Satellite cell regulation

[einstein1905]

I know flexshack asked about this at one time...

Satellite cells are the cells that are "recruited" by IGF-1 to mature and become part of functional muscle fibers, thus increasing the number of "muscle cells", functional ones at least.

It has been shown in rats that overexpression of IGF-1, intramuscularly, promotes great increases in muscle mass and strength for several weeks, but eventually the rate of above average growth plateaus. Researchers hypothesized that this was due to a decreased supply of satellite cells (i.e. the rate of satellite cell mitosis was slower than the rate of IGF-1 mediated satellite cell "recruitment").

What regulates the rate of satellite cell mitosis?.....
One thing to note, in rats overexpressing IGF-1, their GH will be greatly inhibited due to negative feedback by IGF-1 on the hypothalamus, so thier endogenous GH production/release would be nearly nonexistent.

Here are two studies....the first shows that myostatin has a distinct adverse effect on satellite cell replication (mitosis)...the second study shows that GH has a distinct effect on decreasing the expression of myostatin. If you put these two results together, you can conclude that GH has a positive effect on satellite cell mitosis. So, since the mice overespressing IGF-1 had their endogenous GH production suppressed, their rates of satellite cell mitosis was also greatly reduced, and therefore the limiting factor was the inability of rate of satellite cell mitosis to provide an adequate supply of satellite cells for IGF-1 recruitment, so growth eventually stopped.

For those of use using LR3 and noticing gains decreasing after 4-5 weeks, this could very well be due to LR3's suppression of our endogenous GH, which is occuring with use of exogenous IGF-1, and therefore a decreased rate of satellite cell mitosis. The first time I used LR3 I was not using GH at the time, and my gains seemed to slow at the 4 week mark. I'd heard others having the same effects, so I wrote it off as a fact that 4 week cycles were the best, beyond which, gains were harder to achieve. On subsequent cycles of LR3, I have always been using GH with them but have stopped the cycles after 4 weeks due to my earlier conclusion that gains slowed substantially after 4 weeks.
So, JB if you read this, were you using GH at the time you did your 5 week LR3 cycle and continued to get gains?

My theory now is that using GH with LR3 can allow one to cycle for a longer duration with LR3 and continue to gain......I hadn't before given this the chance to work, by cutting my cycles at 4 weeks.

Next cycle will be as long as I continue to see good gains, since I'll be using GH along with it again.

Here are the studies:
J Cell Biol. 2003 Sep 15;162(6):1135-47. Epub 2003 Sep 08. Related Articles, Links


Myostatin negatively regulates satellite cell activation and self-renewal.

McCroskery S, Thomas M, Maxwell L, Sharma M, Kambadur R.

Animal Genomics, AgResearch, Hamilton 2015, New Zealand.

Satellite cells are quiescent muscle stem cells that promote postnatal muscle growth and repair. Here we show that myostatin, a TGF-beta member, signals satellite cell quiescence and also negatively regulates satellite cell self-renewal. BrdU labeling in vivo revealed that, among the Myostatin-deficient satellite cells, higher numbers of satellite cells are activated as compared with wild type. In contrast, addition of Myostatin to myofiber explant cultures inhibits satellite cell activation. Cell cycle analysis confirms that Myostatin up-regulated p21, a Cdk inhibitor, and decreased the levels and activity of Cdk2 protein in satellite cells. Hence, Myostatin negatively regulates the G1 to S progression and thus maintains the quiescent status of satellite cells. Immunohistochemical analysis with CD34 antibodies indicates that there is an increased number of satellite cells per unit length of freshly isolated Mstn-/- muscle fibers. Determination of proliferation rate suggests that this elevation in satellite cell number could be due to increased self-renewal and delayed expression of the differentiation gene (myogenin) in Mstn-/- adult myoblasts. Taken together, these results suggest that Myostatin is a potent negative regulator of satellite cell activation and thus signals the quiescence of satellite cells.


J Clin Endocrinol Metab. 2003 Nov;88(11):5490-6. Related Articles, Links


Myostatin is a skeletal muscle target of growth hormone anabolic action.

Liu W, Thomas SG, Asa SL, Gonzalez-Cadavid N, Bhasin S, Ezzat S.

Department of Medicine, Mount Sinai Hospital and University of Toronto, Ontario, Canada.

Myostatin is a cytokine that has recently been shown to selectively and potently inhibit myogenesis. To investigate the mechanisms of anabolic actions of GH on skeletal muscle growth, we examined the in vitro and in vivo effects of GH on myostatin regulation. Twelve GH-deficient hypopituitary adult subjects were treated with recombinant GH (5 micro g/kg.d) in a double-blind, placebo-controlled fashion. Body composition and physical function were assessed and skeletal muscle biopsies from the vastus lateralis performed at 6-monthly intervals during 18 months of treatment. Myostatin mRNA expression was significantly inhibited to 31 +/- 9% (P < 0.001) of control by GH but not by placebo administration (79 +/- 11%) as determined by quantitative real-time PCR normalized for the housekeeping glyceraldehyde-3-phosphate dehydrogenase gene. The inhibitory effect of GH on myostatin was sustained after 12 and 18 months of GH treatment. These effects were associated with increases in lean body mass and translated into enhanced aerobic performance as determined by maximal oxygen uptake and ventilation threshold. Parallel in vitro studies of skeletal muscle cells demonstrated significant reduction of myostatin expression by myotubes in response to GH, compared with vehicle treatment. Conversely, GH receptor antagonism resulted in up-regulation of myostatin in myoblasts. Given the potent catabolic actions of myostatin, our data suggest that myostatin represents a potential key target for GH-induced anabolism.

[flexshack]

I know flexshack asked about this at one time...

Satellite cells are the cells that are "recruited" by IGF-1 to mature and become part of functional muscle fibers, thus increasing the number of "muscle cells", functional ones at least.

It has been shown in rats that overexpression of IGF-1, intramuscularly, promotes great increases in muscle mass and strength for several weeks, but eventually the rate of above average growth plateaus. Researchers hypothesized that this was due to a decreased supply of satellite cells (i.e. the rate of satellite cell mitosis was slower than the rate of IGF-1 mediated satellite cell "recruitment").

What regulates the rate of satellite cell mitosis?.....
One thing to note, in rats overexpressing IGF-1, their GH will be greatly inhibited due to negative feedback by IGF-1 on the hypothalamus, so thier endogenous GH production/release would be nearly nonexistent.

Here are two studies....the first shows that myostatin has a distinct adverse effect on satellite cell replication (mitosis)...the second study shows that GH has a distinct effect on decreasing the expression of myostatin. If you put these two results together, you can conclude that GH has a positive effect on satellite cell mitosis. So, since the mice overespressing IGF-1 had their endogenous GH production suppressed, their rates of satellite cell mitosis was also greatly reduced, and therefore the limiting factor was the inability of rate of satellite cell mitosis to provide an adequate supply of satellite cells for IGF-1 recruitment, so growth eventually stopped.

For those of use using LR3 and noticing gains decreasing after 4-5 weeks, this could very well be due to LR3's suppression of our endogenous GH, which is occuring with use of exogenous IGF-1, and therefore a decreased rate of satellite cell mitosis. The first time I used LR3 I was not using GH at the time, and my gains seemed to slow at the 4 week mark. I'd heard others having the same effects, so I wrote it off as a fact that 4 week cycles were the best, beyond which, gains were harder to achieve. On subsequent cycles of LR3, I have always been using GH with them but have stopped the cycles after 4 weeks due to my earlier conclusion that gains slowed substantially after 4 weeks.
So, JB if you read this, were you using GH at the time you did your 5 week LR3 cycle and continued to get gains?

My theory now is that using GH with LR3 can allow one to cycle for a longer duration with LR3 and continue to gain......I hadn't before given this the chance to work, by cutting my cycles at 4 weeks.

Next cycle will be as long as I continue to see good gains, since I'll be using GH along with it again.

Here are the studies:
J Cell Biol. 2003 Sep 15;162(6):1135-47. Epub 2003 Sep 08. Related Articles, Links


Myostatin negatively regulates satellite cell activation and self-renewal.

McCroskery S, Thomas M, Maxwell L, Sharma M, Kambadur R.

Animal Genomics, AgResearch, Hamilton 2015, New Zealand.

Satellite cells are quiescent muscle stem cells that promote postnatal muscle growth and repair. Here we show that myostatin, a TGF-beta member, signals satellite cell quiescence and also negatively regulates satellite cell self-renewal. BrdU labeling in vivo revealed that, among the Myostatin-deficient satellite cells, higher numbers of satellite cells are activated as compared with wild type. In contrast, addition of Myostatin to myofiber explant cultures inhibits satellite cell activation. Cell cycle analysis confirms that Myostatin up-regulated p21, a Cdk inhibitor, and decreased the levels and activity of Cdk2 protein in satellite cells. Hence, Myostatin negatively regulates the G1 to S progression and thus maintains the quiescent status of satellite cells. Immunohistochemical analysis with CD34 antibodies indicates that there is an increased number of satellite cells per unit length of freshly isolated Mstn-/- muscle fibers. Determination of proliferation rate suggests that this elevation in satellite cell number could be due to increased self-renewal and delayed expression of the differentiation gene (myogenin) in Mstn-/- adult myoblasts. Taken together, these results suggest that Myostatin is a potent negative regulator of satellite cell activation and thus signals the quiescence of satellite cells.


J Clin Endocrinol Metab. 2003 Nov;88(11):5490-6. Related Articles, Links


Myostatin is a skeletal muscle target of growth hormone anabolic action.

Liu W, Thomas SG, Asa SL, Gonzalez-Cadavid N, Bhasin S, Ezzat S.

Department of Medicine, Mount Sinai Hospital and University of Toronto, Ontario, Canada.

Myostatin is a cytokine that has recently been shown to selectively and potently inhibit myogenesis. To investigate the mechanisms of anabolic actions of GH on skeletal muscle growth, we examined the in vitro and in vivo effects of GH on myostatin regulation. Twelve GH-deficient hypopituitary adult subjects were treated with recombinant GH (5 micro g/kg.d) in a double-blind, placebo-controlled fashion. Body composition and physical function were assessed and skeletal muscle biopsies from the vastus lateralis performed at 6-monthly intervals during 18 months of treatment. Myostatin mRNA expression was significantly inhibited to 31 +/- 9% (P < 0.001) of control by GH but not by placebo administration (79 +/- 11%) as determined by quantitative real-time PCR normalized for the housekeeping glyceraldehyde-3-phosphate dehydrogenase gene. The inhibitory effect of GH on myostatin was sustained after 12 and 18 months of GH treatment. These effects were associated with increases in lean body mass and translated into enhanced aerobic performance as determined by maximal oxygen uptake and ventilation threshold. Parallel in vitro studies of skeletal muscle cells demonstrated significant reduction of myostatin expression by myotubes in response to GH, compared with vehicle treatment. Conversely, GH receptor antagonism resulted in up-regulation of myostatin in myoblasts. Given the potent catabolic actions of myostatin, our data suggest that myostatin represents a potential key target for GH-induced anabolism.

well put einstein. please keep me updated with the results of your upcoming experiment. i'm very interested. i'm also curious about the question you asked JB.

one thing to mention though, is that when using lr3, one's nat. gh production isn't going to be "shut down" for the duration of the cycle (especially if one is administering it only once daily). maybe it's an issue of the length of time throughout the day that gh is being suppressed that is affecting myostatin? maybe if we cycle lr3 differently, we could possibly extend its length of effectiveness (eod)?

another idea to consider is blocking myostatin through methods/supps/drugs other than gh. to be honest, i don't know much on myostatin, nor do i know if what i suggested above is even feasible. if i remember correctly, i think i've heard of some supplements that claim to do this, but i also remember hearing that they're garbage.

[einstein1905]

well put einstein. please keep me updated with the results of your upcoming experiment. i'm very interested. i'm also curious about the question you asked JB.

one thing to mention though, is that when using lr3, one's nat. gh production isn't going to be "shut down" for the duration of the cycle (especially if one is administering it only once daily). maybe it's an issue of the length of time throughout the day that gh is being suppressed that is affecting myostatin? maybe if we cycle lr3 differently, we could possibly extend its length of effectiveness (eod)?

another idea to consider is blocking myostatin through methods/supps/drugs other than gh. to be honest, i don't know much on myostatin, nor do i know if what i suggested above is even feasible. if i remember correctly, i think i've heard of some supplements that claim to do this, but i also remember hearing that they're garbage.

No, you're right about our nat. GH not being completely suppressed while using LR3 (especially 1x/day), but there is suppression nonetheless, and adding exogenous GH in supraphysiological amounts should have quite an impact on satellite cell mitosis, especially relative to the levels of GH we'd be producing during an LR3 cycle.

As for myostatin, there are clinical trials going on now with a neutralizing antibody to myostatin, called MYO-029....we'll see where that goes.

[flexshack]

As for myostatin, there are clinical trials going on now with a neutralizing antibody to myostatin, called MYO-029....we'll see where that goes.

why is it even being made? there are medical reasons for suppressing myostatin? maybe in degenerative muscle wasting diseases and such?

[einstein1905]

why is it even being made? there are medical reasons for suppressing myostatin? maybe in degenerative muscle wasting diseases and such?

Primarily to treat muscular dystrophy and muscle wasting associated with cancer and AIDS

[flexshack]

i just found this on blocking myostatin. it also answers my last question above.

http://www.mdausa.org/news/021127micedmd.html

also, paragraphs 8 and 9 caught my attention. i believe it pertains to our discussion on satellite cells. i don't see why he claims that the accelerated recruitment could cause worsening of the muscular dystrophy? do you? and i wonder if the reason and negative effects would also pertain to normal individuals?

[flexshack]

Primarily to treat muscular dystrophy and muscle wasting associated with cancer and AIDS

woops! you beat me to it, lol!

[einstein1905]

i just found this on blocking myostatin. it also answers my last question above.

http://www.mdausa.org/news/021127micedmd.html

also, paragraphs 8 and 9 caught my attention. i believe it pertains to our discussion on satellite cells. i don't see why he claims that the accelerated recruitment could cause worsening of the muscular dystrophy? do you? and i wonder if the reason and negative effects would also pertain to normal individuals?

I'll look into it more, but myostatin keeps satellite cells in a quiescent state, meaning nonmitotic. It's often the case that when cells are in a state of quiescence, they don't express many of the proteins that are expressed when they are actively dividing. one of these proteins could very well be IGF receptors. So, if myostatin was substantially inhibited, the majority of satellite cells would be mitotic and expressing IGF receptors and therefore be susceptible for recruitment and maturation, which could expedite their depletion. I'd say it's all going to be relative and dose-dependent....e.g. a 30% reduction in myostatin we'll just say increases mitotic activity in satellite cells by 30%, which may be not enough to deplete the supply of satellite cells for someone using 40mcg/day of LR3, but if myostatin were inhibited 50%, this may be enough to cause depletion, however, then the dose of LR3 could also be reduced to accommodate.

I'd say it's just speculative on their part.....just stating a potential risk, but who knows?

[flexshack]

I'll look into it more, but myostatin keeps satellite cells in a quiescent state, meaning nonmitotic. It's often the case that when cells are in a state of quiescence, they don't express many of the proteins that are expressed when they are actively dividing. one of these proteins could very well be IGF receptors. So, if myostatin was substantially inhibited, the majority of satellite cells would be mitotic and expressing IGF receptors and therefore be susceptible for recruitment and maturation, which could expedite their depletion. I'd say it's all going to be relative and dose-dependent....e.g. a 30% reduction in myostatin we'll just say increases mitotic activity in satellite cells by 30%, which may be not enough to deplete the supply of satellite cells for someone using 40mcg/day of LR3, but if myostatin were inhibited 50%, this may be enough to cause depletion, however, then the dose of LR3 could also be reduced to accommodate.

but then wouldn't your theory of using gh (to inhibit myostatin) with lr3 cause the same complicated dose-dependent issue?

I'd say it's just speculative on their part.....just stating a potential risk, but who knows?

when speaking about degenerative diseases such as the one in the article, i don't even understand the risk of depleting the satellite cells. how could the depletion worsen the dystrophy if the cells are being recruited to form more muscle! where's the problem or "worsening" there? b/c i'm not seeing it. maybe i'm just misunderstanding this?

[einstein1905]

when speaking about degenerative diseases such as the one in the article, i don't even understand the risk of depleting the satellite cells. how could the depletion worsen the dystrophy if the cells are being recruited to form more muscle! where's the problem or "worsening" there? b/c i'm not seeing it. maybe i'm just misunderstanding this?

I'd assume they're referring to more long term. If the satellite cell population is depleted, even though this contributed to extensive regeneration of muscle, the causative agent still remains and confers disease, in the case of DMD, a genetic defect. So, inevitably the muscle that has been regenerated is still doomed to the same fate as the muscle it replaced.




Myogenic satellite cells: physiology to molecular biology
Thomas J. Hawke1 and Daniel J. Garry1,2
J Appl Physiol 91: 534-551, 2001;
8750-7587/01


"Most myopathies have a molecular mutation that affects the structural or cytoskeletal proteins in skeletal muscle. Duchenne muscular dystrophy (DMD) is the most common and the most devastating of the muscular dystrophies (20, 55, 76, 82, 127). Disease progression and death are ultimately due to a failure of the myogenic satellite cells to maintain muscle regeneration (36, 80). DMD is a recessive X-linked disease that results in a null mutation at the dystrophin locus (20, 82, 127). The absence of this cytoskeletal protein renders the muscle fiber extremely fragile. In response to mechanical stress associated with repeated contraction, there is widespread degeneration. The satellite cells respond to the injury by repopulating the injured skeletal muscle with defective myofibers lacking dystrophin. This process results in continuous degeneration-regeneration cycles and ultimately exhausts the satellite cell pool (36, 80).

Clinical symptoms are apparent by 4-5 yr of age in boys with DMD (10, 20, 127). DMD patients (4-5 yr of age) have been shown to undergo more skeletal muscle regeneration than that measured in a total of six normal patients over 60 yr of age (46). These results were confirmed by Renault et al. (151), who demonstrated that the proliferative life span of satellite cells derived from a 9-yr-old DMD patient was approximately one-third of an age-matched control. Proliferative fatigue or senescence of the satellite cell population and the milieu of the DMD skeletal muscle may collectively impair the proliferative or regenerative capacity of this cell population. In the DMD patient, increased levels of IGF binding proteins (IGFBP) are released by fibroblasts. The elevated IGFBP sequesters IGF-I, limiting its bioavailability for satellite cells and ultimately resulting in increased skeletal muscle fibrosis (120). The evidence to date demonstrates the tremendous strain and ultimate failure of the satellite cell population to adequately compensate for the persistent degeneration-regeneration process that is occurring in the DMD skeletal muscle."

[einstein1905]

Here is another study showing testosterone having a role in satellite cell activathion. However, the study is generalized and doesn't look at specifics. We know that testosterone increases GH, which increases IGF-1, so really all this study says to me is .....what I just said, but AR has lots of bandwidth, so I'm posting it anyway.

Dev Biol. 1995 May;169(1):286-94. Related Articles, Links


Testosterone treatment results in quiescent satellite cells being activated and recruited into cell cycle in rat levator ani muscle.

Joubert Y, Tobin C.

Laboratoire de Neurobiologie des signaux intercellulaires, URA 1488, Paris.

At puberty, the male levator ani (LA) muscle exhibits muscle fiber hypertrophy. This fiber enlargement can be provoked in the female LA muscle by testosterone treatment. In both cases the hypertrophic process is accompanied by an increase in the number of satellite cells and myonuclei. The present ultrastructural autoradiographic study was undertaken in order to investigate (1) whether satellite cells when stimulated by testosterone can undergo DNA synthesis by incorporating [3H]thymidine ([3H]Tdr); (2) whether the whole satellite cell population is committed in the cell cycle; and (3) whether the new myonuclei originate from fusion of the satellite cells with the preexistant myofibers. Thirty-day-old female rats treated with a single testosterone injection received a single injection of [3H]Tdr at 24, 28, 32, to 34 hr after testosterone treatment. LA muscles were removed 2 hr after [3H]Tdr injection. This first series of experiments allowed us to determine that onset of DNA synthesis in satellite cells occurs within the 34th and the 36th hour after testosterone treatment. To obtain a more precise timing, 30-day-old female rats treated with a single testosterone injection received multiple injections every 30 min either from Hour 32 to 33.5 or from Hour 34 to 35.5 LA muscles were removed 30 min after the last [3H]Tdr injection. This showed that the onset of satellite cell replication occurred between the 32nd and the 34th hour after testosterone treatment. However, only 30% of the satellite cell population was affected by this proliferative process regardless of the experimental protocol used. To confirm that the increase in myonuclei number results from incorporation of satellite cells into mature myofibers, 30-day-old female rats treated with a single injection of testosterone received a single injection of [3H]Tdr on the 60th hour after testosterone treatment. LA muscles were removed at 63, 84, to 108 hr after [3H]Tdr injection. We conclude that testosterone induces satellite cell proliferation at around the 33rd hour and that the increased number of myonuclei reported in our previous study is due to fusion of labeled satellite cells with myofibers.


Here is the Figure legend for the attached image:
Fig. 4. Satellite cell response to myotrauma. *Skeletal muscle trauma or injury may be minor (e.g., resistance training) or may be more extensive (e.g., toxin injection, Duchenne muscular dystrophy). In response to an injury, satellite cells become activated and proliferate. Some of the satellite cells will reestablish a quiescent satellite cell pool through a process of self-renewal. Satellite cells will migrate to the damaged region and, depending on the severity of the injury, fuse to the existing myofiber or align and fuse to produce a new myofiber. In the regenerated myofiber, the newly fused satellite cell nuclei will initially be centralized but will later migrate to assume a more peripheral location.

[einstein1905]

J. Anim. Sci. 2003. 81:965-972
© 2003 American Society of Animal Science

Growth factor messenger RNA levels in muscle and liver of steroid -implanted and nonimplanted steers1,2
M. E. White, B. J. Johnson, M. R. Hathaway* and W. R. Dayton*,3
* Animal Growth and Development Laboratory, Department of Animal Science, University of Minnesota, St. Paul 55108 and Department of Animal Sciences and Industry, Kansas State University, Manhattan 66506





Ribonuclease protection assays were used to measure steady-state semimembranosus muscle and/or hepatic levels of IGF-I, IGFBP-3, IGFBP-5, hepatocyte growth factor (HGF), and myostatin messenger RNA (mRNA) in steers implanted from 32 to 38 d with Revalor-S, a combined trenbolone acetate and estradiol implant. Insulin -like growth factor-I mRNA levels were 69% higher (P < 0.01, n = 7) in the livers of implanted steers than in the livers of nonimplanted steers. Similarly, IGF-I mRNA levels were 50% higher (P < 0.05, n = 7) in the semimembranosus muscles of implanted steers than in the same muscles from nonimplanted steers. Hepatic IGFBP-3 mRNA levels were 24% higher (P < 0.07, n = 7) in implanted steers than in nonimplanted steers. Hepatic HGF and IGFBP-5 mRNA levels did not differ between implanted and nonimplanted steers. Similarly, muscle IGFBP-3, IGFBP-5, HGF, and myostatin mRNA levels were not affected by treatment. Previous data from these same steers have shown that circulating IGF-I and IGFBP-3 concentrations were 30 to 40% higher (P < 0.01, n = 7) in implanted steers than in nonimplanted, control steers. Additionally, the number of actively proliferating satellite cells that could be isolated from the semimembranosus muscle was 45% higher (P < 0.01, n = 7) for implanted steers than for nonimplanted steers. Viewed together, these data suggest that increased muscle IGF-I levels stimulate increased satellite cell proliferation, resulting in the increased muscle growth observed in Revalor-S implanted steers.




Feedlot steers implanted with Revalor-S, a combined implant containing 120 mg of trenbolone acetate (TBA) and 24 mg of estradiol (E2), exhibited a 20 to 25% increase in rate of gain, a 15 to 20% increase in feed efficiency, increased carcass protein, and increased longissimus muscle area compared with nonimplanted steers (Johnson et al., 1996a). Implantation of feedlot steers with Revalor-S also resulted in an increase in the number of actively proliferating satellite cells that could be isolated from the semimembranosus muscle (SM) (Johnson et al., 1998a). Steady-state IGF-I messenger RNA (mRNA) levels were increased in the longissimus muscle of implanted steers (Johnson et al., 1998b) and IGF-I levels were increased in the diaphragm muscle of nandrolone -treated rats (Lewis et al., 2002). Additionally, virally induced overexpression of IGF-I in muscle tissue resulted in a 15% increase in muscle mass in young adult mice (Barton-Davis et al., 1998), and overexpression of IGF-I extends the replicative lifespan of satellite cells (Barton-Davis et al., 1999; Chakravarthy et al., 2000). Viewed together, these data suggest that steroid-induced muscle growth may result from increased muscle IGF-I levels that maintain satellite cells in a proliferative state as steers approach maturity. Additionally, IGFBP (James et al., 1993; Damon et al., 1998; Yang et al., 1999), hepatocyte growth factor (HGF) (Allen et al., 1995; Tatsumi et al., 1998; Sheehan and Allen, 1999), or myostatin could play a role in anabolic steroid-induced muscle growth (McPherron et al., 1997; Carlson et al., 1999; Sakuma et al., 2000). Consequently, we have measured steady-state HGF, myostatin, IGF-I, IGFBP-3, and IGFBP-5 mRNA levels in SM and IGF-I, IGFBP-3, IGFBP-5, and HGF mRNA levels in livers from Revalor-S-treated steers and nonimplanted steers.

[einstein1905]

Am J Physiol Endocrinol Metab 281: E1159-E1164, 2001;
0193-1849/01 $5.00
Vol. 281, Issue 6, E1159-E1164, December 2001

Comparison of GH, IGF-I, and testosterone with mRNA of receptors and myostatin in skeletal muscle in older men
Taylor J. Marcell1, S. Mitchell Harman1, Randall J. Urban3, Daniel D. Metz1, Buel D. Rodgers2, and Marc R. Blackman2
1 Intramural Research Program, National Institute on Aging, National Institutes of Health; 2 Division of Endocrinology and Metabolism, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21224; and 3 Department of Internal Medicine, University of Texas Medical Branch, Galveston, Texas 77555



Growth hormone (GH), insulin -like growth factor I (IGF-I), and testosterone (T) are important mediators of muscle protein synthesis, and thus muscle mass, all of which decline with age. We hypothesized that circulating hormones would be related to the transcriptional levels of their respective receptors and that this expression would be negatively related to expression of the myostatin gene. We therefore determined content of mRNA transcripts (by RT-PCR) for GH receptor (GHR), IGF-I, androgen receptor (AR), and myostatin in skeletal muscle biopsy samples from 27 healthy men >65 yr of age. There were no significant relationships between age, lean body mass, or percent body fat and transcript levels of GHR, IGF-I, AR, or myostatin. Moreover, there were no significant correlations of serum GH, IGF-I, or T with their corresponding target mRNA levels (GHR, intramuscular IGF-I, or AR) in skeletal muscle. However, GHR was negatively correlated (r = 0.60, P = 0.001) with myostatin mRNA levels. The lack of apparent relationships of muscle transcripts with their respective ligands in healthy older adults suggests that age-related deficits in both GH and T may lead to an increase in myostatin expression and a disassociation in autocrine IGF-I effects on muscle protein synthesis, both of which could contribute to age-related sarcopenia.



IN HEALTHY YOUNG ADULTS, under equilibrium conditions, skeletal muscle protein synthesis and degradation are a balanced, dynamic process with no net change occurring in skeletal muscle mass. During aging, however, muscle tissue is gradually lost, often resulting in diminished mass and strength, a condition referred to as sarcopenia, which contributes to frailty. This loss of muscle results from a net imbalance between the rates of protein synthesis and degradation (22). Protein synthesis can be stimulated by various signals, including hormones, metabolic demand, and functional overload (10, 23). Protein breakdown is also under the influence of these same factors, which stimulate lysosomal and ubiquitin degradation processes. Insulin, growth hormone (GH), insulin-like growth factor I (IGF-I), and testosterone (T) are anabolic hormones, all of which increase muscle mass by stimulating protein synthesis and/or inhibiting protein breakdown, whereas cortisol is a potent stimulus to protein catabolism (6, 9, 22). Circulating levels of these various hormones are altered by the aging process, potentially contributing to sarcopenia (2, 5, 12, 27).

Although GH-induced IGF-I production in the liver is the major source of circulating IGF-I and mediates many GH metabolic effects, local IGF-I production within target tissues, under the influence of both GH and T (28), accounts for >50% of total IGF-I production and appears to be more important for stimulating muscle growth and repair (10, 14, 16). How changes in hormone levels with age affect the IGF-I pathway in skeletal muscle remains poorly understood.

Myostatin, a recently discovered member of the transforming growth factor (TGF)- superfamily, is an autocrine factor that is a potent inhibitor of muscle development (20). Postnatally, myostatin is expressed in varying levels exclusively in skeletal muscle (4, 29) and preferentially in fast-type skeletal muscle fibers (4). Age-related loss of skeletal muscle is associated with a selective atrophy of the fast-type skeletal muscle fibers (17). Thus, whether the age-related decline in anabolic hormonal status allows a progressive increase in myostatin expression that would contribute to sarcopenia is an important question (15). However, by what mechanism hormones interact with myostatin remains unclear.

Therefore, in the current study, we measured plasma levels of GH, IGF-I, T, and cortisol and, using the reverse transcription-polymerase chain reaction (RT-PCR), determined gene expression of IGF-I and myostatin in skeletal muscle biopsy samples from healthy older men. Because circulating hormones can autoregulate their actions by effects on their own receptors (3), we also determined mRNA levels for GH receptor (GHR) and androgen receptor (AR). The present study was designed to test the hypotheses that 1) circulating anabolic hormone concentrations are significantly related to the transcriptional levels of their respective receptors, 2) the expression of GHR, IGF-I, and AR is up- or downregulated coordinately, and 3) the expression of anabolic hormone receptors is negatively related to expression of the myostatin gene.

[einstein1905]

Advanced Training Planning for Bodybuilders: Part 2
written by Bryan Haycock


Introduction
In part one of this article we discussed the history of resistance training and some very basic principles at work during adaptation to resistance training. Specificity was outlined as a governing principle with which we can predict the outcomes of our training. Low volume/high load training produce increases in neuromuscular efficiency and motor unit recruitment, while high volume/moderate load training produces only moderate increases in strength and neuromuscular adaptations along with marked hypertrophy of both slow and fast twitch fibers. Also discussed were issues such as rational and irrational adaptation. Dramatic increases in sarcomere volume without increases in myo-nuclear number, seen during irrational adaptation, effectively inhibits further increases in the production of contractile proteins and diminishes recovery and performance. Slower increases in sarcomere volume, as seen in rational adaptation, actually facilitates recovery and leads to a more steady increase in both size and strength. In part two we will discuss the mechanisms responsible for the specific nature of adaptation and look at ways of applying this knowledge to build size and strength.

The Stimulus for Muscle Growth
To most people, the way to get a muscle to grow larger is simply a matter of "exercising" the muscle you want to grow. This is a very simplistic way of looking at it. Those individuals who fail to realize that there is a right way and a wrong way to train a muscle never go on to develop out of the ordinary physiques. Don't get me wrong, the knowledge of how muscle tissue grows in response to training is not a requirement to be a successful bodybuilder. Anabolics have become the ultimate "cheat sheet". They effectively reduce the overload threshold necessary for compensatory hypertrophy and elevate the genetic limit or plateau. If you chose not to use potent synthetic hormones you are in for a much more difficult road, and a good understanding of muscle hypertrophy will be invaluable to you as you train over the years.

At the very foundation of muscle hypertrophy is load induced tissue strain. Muscle tissue must undergo mechanical strain in order to begin the biochemical steps necessary for adaptive growth (Clarke,1996). This "strain" induced by the loading of the tissue leads to sarcolemma microtrauma. The wounding of the muscle cell after training is characterized by myofibrillar disruption, Z line smearing, discontinuity of sarcomeres and an increase in the porosity, or permeability, of the sarcolemma. The effect or result of this nonfatal cellular damage is the production, and release of growth factors that then interact with the damaged cell itself and also very importantly, with satellite cells.

Satellite cells are myogenic stem cells that serve to assist postnatal growth and regeneration in adult skeletal muscle. Following proliferation (reproduction) and subsequent differentiation (to become a specific type of cell), these satellite cells will fuse with one another or with the adjacent damaged muscle fiber, thereby increasing myonuclei numbers for fiber growth and repair. Proliferation is necessary in order to meet the needs of thousands of muscle cells all potentially requiring additional nuclei. Differentiation is necessary in order for the new nucleus to behave as a nucleus of muscle origin. In order to better understand what is physically happening between satellite cells and muscle cells, try to picture 2 oil droplets floating on water. The two droplets represent a muscle cell and a satellite cell. Because the lipid bilayer of cells are hydrophobic just like common oil droplets, when brought into proximity to one another in an aqueous environment, they will come into contact for a moment and then fuse together to form one larger oil droplet. Now whatever (i.e. nuclei) was dissolved within one droplet will then mix with the contents of the other droplet. This is a simplified model of how satellite cells donate nuclei to existing muscle cells.

There appears to be a finite limit placed on the cytoplasmic/nuclear ratio (Rosenblatt,1994). Whenever a muscle grows in response to functional overload there is a positive correlation between the increase in the number of myonuclei and the increase in fiber cross sectional area (CSA). When satellite cells are prohibited from donating viable nuclei, overloaded muscle will not grow (Rosenblatt,1992; Phelan,1997). It is not a stretch to say that satellite cell activity is a required step, or prerequisite, in compensatory muscle hypertrophy, for without it, a muscle simply cannot significantly increase total protein content nor CSA.

Some factors which regulate this process are exercise, trauma, passive stretch, massage, innervation, and the activity of soluble growth factors. Three classes of growth factors in particular have been studied extensively with respect to their effects on satellite cell proliferation and differentiation in vitro. They are; fibroblast growth factors (FGF), insulin -like growth factors (IGF), and the transforming growth factor-beta superfamily (TGF-beta). When administered in combination, these factors can induce satellite cell activities in vitro which mimic those typical of satellite cells found in vivo in growing, regenerating, or healthy mature muscle. In essence they can mimic the effects of loading without any microtrauma actually being done.

FGF is one growth factor being actively studied. Recent studies have shown that an increase in the permeability of the sarcolemma is necessary for the release of fibroblast growth factor (FGF). This is because FGF does not contain a signal transduction peptide sequence (Abraham,1986) and thus does not exit the source cell through vesicle mediated exocytosis. This "signal transduction peptide sequence" is necessary for the protein to be incorporated into intracellular vesicles and actively transported to the cell surface, and then ultimately released. Instead, it must be able to pass directly through the phospholipid bilayer of the wounded sarcomere in order to have an autocrine and paracrine effect on target tissues. This is accomplished in adult muscle fibers by inflicting microtrauma.

FGF, specifically FGF-beta, has been shown to stimulate proliferation but suppress differentiation of myogenic stem cells. This has the result of increasing the availability of satellite cells but reduces the number of them that are actively fusing with nearby muscle cells. In a study where FGF was injected directly into healthy muscle tissue, it had the effect of increasing muscle DNA content and IGF-1 peptides, but had no significant effect on total protein content or gross muscle weight (Adams,1998). These researchers speculate that the increase in DNA content was the result of increased satellite cell number. The fact that FGF was unable to induce hypertrophy reflects the fact that FGF inhibits satellite cell differentiation (i.e. the satellite cells never actually became of the muscle cell type) preventing the nuclei from producing muscle specific proteins and thereby short circuiting a critical step the growth process. An additional study involving both local injection and local implantation of FGF pellets into "regenerating", or damaged muscle, failed to show any effect of FGF on the ability of muscle tissue to regenerate after microtrauma (Mitchell,1996). In this study FGF did have the effect of enhancing satellite cell proliferation as well as angiogenesis (capillary formation) nevertheless, it appears that FGF is not the limiting factor in compensatory muscle hypertrophy.

Insulin-like growth factor (specifically IGF-1) stimulates both proliferation and differentiation in an autocrine-paracrine manner, although it induces differentiation to a much greater degree. IGF-1, when injected locally, increases satellite cell activity, muscle DNA, muscle protein content, muscle weight and muscle cross sectional area (Adams,1998). As discussed earlier, the proliferation and differentiation of satellite cells is critical part of compensatory hypertrophy. The importance of IGF-1 lies in the fact that all of its apparent functions act to induce muscle growth with or without overload although it really shines as a growth promoter when combined with physical loading of the muscle.

IGF-1 also acts as an endocrine growth factor having an anabolic effect on distant tissues once released into the blood stream by the liver. In human volunteers, detailed information on the effect of IGF-1 on protein synthesis, degradation and balance has been obtained by using the arteriovenous difference of labeled and unlabeled phenylalanine across the forearm (Barrett and Gelfand 1989). In the aforementioned studies, "systemic" IGF-1 infusion for 6 h caused positive amino acid balance, both by inhibiting protein degradation and stimulating protein synthesis (Fryburg, 1994). This differs from the effect of peptide "hormones" such as insulin, which does not stimulate synthesis in adults (Bennet et al. 1990, Fryburg 1990, Gelfand and Barrett 1987, McNurlan, 1994). Therefore IGF-1 possesses the insulin-like property of inhibiting degradation, but in addition can stimulate protein synthesis (Fryburg 1991). The insulin-like effects are probably due to the similarity of the signaling pathways between insulin and IGF-1 following ligand binding at the receptors (Schumacher 1991, Gual 1998).

The ability of IGF-I to stimulate protein synthesis resembles the action of GH, which was shown in separate studies on volunteers to stimulate protein synthesis without affecting protein degradation (Fryburg et al. 1991, Fryburg and Barrett 1993). Although it is often believed that the effects of GH are mediated through IGF-1, this cannot be the case entirely. First, the effects of the two hormones were different, in that GH did not change protein degradation. Second, the effect of GH was observed with little or no change in systemic IGF-1 and GH concentrations because the GH was infused directly into the brachial artery (Fryburg et al. 1991).

Transforming growth factor-beta (TGF-beta) slightly depresses proliferation but inhibits differentiation. So although it is called a growth factor, it is an "inhibitory" factor involved in muscle growth. For example, one TGF-beta member known as growth/differentiation factor-8 (GDF-8), was found to have profound effects on muscle growth (McPherron and Lawler,1997). GDF-8 was found to exist in many muscles throughout the body. In order to identify the function of this protein, researchers "targeted", or "knocked out" the gene responsible for producing it. The result was nothing less than miraculous. The mice who lacked the gene went on to grow muscles up to 3 times larger than normal mice. This is a 300% increase in muscle weight with no specific exercise or mechanical loading! You may remember pictures of these mice published in issue number 188 in MuscleMag International (MuscleMag,1998). The increase in the mass of the "mutant" mice muscles was a result of both increased hypertrophy and hyperplasia (an increase in fiber number). There are also naturally occurring mutations to this gene that result in "double muscling" of animals such as livestock. Two breeds of cattle known as Belgian Blue and Piedmontese exhibit naturally occurring mutations on the myostatin gene (McPherron and Lee,1997). This changes the amino acid sequence of the GDF-8 peptide and greatly attenuates its physiological activity.

All of these growth factors are brought into play in human models of compensatory hypertrophy. In summary, resistance exercise of sufficient load and volume causes microtrauma to the sarcolemma which increases the release and production of growth factors. This is done through increased permeability of the wounded cell membrane allowing soluble growth factors to "leak" out into the intercellular space. The growth factors then go on to increase the number, or "proliferation" of satellite cells, also called myogenic stem cells. This is done through interaction with growth factor receptors on the surface of these cells. These growth factors then go further to induce the conversion, or "differentiation" of these undifferentiated cells into cells expressing DNA of muscle origin. Once these satellite cells have undergone differentiation they can then fuse to existing muscle cells. This fusion allows the satellite cell to donate needed myonuclei to the wounded or developing muscle cell. The effect of increasing the number of nuclei within a cell allows for increased protein synthesis and ultimately hypertrophy. As mentioned earlier, there is a nuclear to cytoplasmic ratio, or "nuclear domain", that is closely regulated by the cell. If the muscle cell is prevented from increasing the number of nuclei, it does not grow in response to overload (Phelan,1997).

All of the aforementioned processes occur with or without systemic hormonal influence or nutritional abundance (Borer,1995). In this way the hypertrophic response can be limited to the overloaded tissue. I do not mean to imply that exercise induced growth is not effected by systemic hormones and/or nutritional abundance, only that mechanisms are in place that allow for growth in localized muscle tissue in the absence of endocrine support and adequate nutritional status.

The Stimulus for Strength
The foundation for the development of strength is neuromuscular in nature. Increases in strength from resistance exercise has been attributed to several neural adaptations including altered recruitment patterns, rate coding, motor unit synchronization, reflex potentiation, prime mover antagonist activity, and prime mover agonist activity. Aside from incremental changes in the number of contractile filaments, voluntary force production is largely a matter of "activating" motor units. In order to ascertain the relative contribution of each of these mechanisms, various measurement techniques have been utilized. Hereafter we will briefly discuss each of these mechanisms as they relate to resistance training.

Recruitment of motor units can be measured with Electromyography (EMG). As a muscle contracts, the electrical signal initiated by the motor nerve can be detected with EMG. The intensity or magnitude of this signal is sometimes described as "neural drive". In order to explain increases in strength from resistance exercise, researchers have measured the changes in EMG activity in weight training subjects.

Hakkinen and co-workers have shown that there is an increase in EMG activity with strength training as well as a decrease in EMG activity upon cessation of training (Hakkinen,1983). Fourteen male subjects (20-30 yr) accustomed to weight training went through progressive strength training of combined concentric and eccentric contractions three times per week for 16 wk. The active training period was followed by an eight week detraining period. The training program consisted mainly of dynamic exercises for leg extensors with the loads of 80-120% of one maximum concentric repetition (1RM). Significant improvements in muscle function were observed in early conditioning; however, the increase in maximal force during the very late training period was greatly limited. Marked improvements in muscle strength were accompanied by significant increases in the neural activation (EMG) of the leg extensor muscles. The relationship between EMG and high absolute forces changed during the training period. The occurrence of these changes varied during the course of training. During detraining, there was a decline in EMG activity.

Now those who would argue that increases in strength are solely due to increased recruitment of motor units would have a difficult time defending themselves in light of other research. The is a method of measuring motor unit activity called "Interpolated Twitch Technique", or ITT. ITT is used to determine the extent of activation of the entire muscle. Merton (Merton, 1954) was the first to use this technique to describe whole muscle activation. He showed full activation of the adductor pollicis with fatigue in untrained subjects. Several other studies have since shown a similar ability of untrained subjects to voluntarily fully activate various muscle groups (Bellemare 1983, Chapman 1985, Gandevia 1988, Belanger 1981). This directly contradicts the theory of strength increases due to the ability to activate more motor units.

The activation of motor units is done in an asynchronous fashion, meaning that not all fibers contract at the same time within a given muscle. There is a hierarchy to the order of fiber recruitment in muscle tissue. Because fiber activation is not "analog" or variable in nature, in other words, a fiber is either fully activated or fully quiescent, the brain must control contraction intensity by altering the number of fibers it activates. In general, slow twitch fibers are activated first followed by larger fast twitch fibers. Now when muscles begin to fatigue the asynchronous firing of fibers become more and more synchronized (Butchal, 1950). This allows for greater force production. This synchronization of muscle fibers has been linked to increases in voluntary strength (Milner-Brown, 1975). Now although increases in motor unit synchronization have been reported with training, studies involving artificial stimulation show that force development with asynchronous stimulation is greater and smoother (Clamann, 1988). In addition, researchers have shown that the rate of force development in brief maximal contractions is faster in voluntary than in evoked contractions (Miller, 1981). So from these studies we see that although synchronization of motor units can increase with training, asynchronous motor unit activation is more advantageous to rate and magnitude of force development than is synchronous activation.

Increases in "reflex potentiation" have also been linked to resistance training (Sale and Upton 1983, Sale and MacDougall 1983) as well as decreases with immobilization (Sale, 1982). The actual benefit, if any, of this adaptation is unclear. An increase in reflex potentiation would contribute to the voluntary EMG signal augmenting the motor or neuronal drive. Nevertheless, because untrained individuals have been shown to be able to fully recruit their motor units, the purpose of increased reflex potential remains undecided.

Finally, that activity of prime mover agonists and antagonists plays a role in directed voluntary strength. The obvious role of agonists is to assist the prime mover by guidance and stabilization. This could be termed "coordination". It is well known that any unaccustomed exercise requires practice in order to develop sufficient coordination to allow maximum efficiency of muscular effort. The role of antagonistic muscle groups is more complicated. They serve to prevent damage through co-contraction as well as ensure less resistance through relaxation to prime mover contractility.

The protective mechanisms function by way of golgi tendon organs (GTO). The GTO is sensitive to force output or tension within the muscle. They are located at the musculo-tendonous junction and is contained within a compressible collagenous capsule. Fibers of the GTO are connected directly to muscle fibers as well as to Type "Ib" inhibitory neurons within the muscle. The physical structure of the GTO allows it to be sensitive to stretch or load present in the muscle. Think of the notorious "Chinese finger trap". You first stick you fingers in each end. Then as you pull your fingers apart, the structure of the woven tube causes it shrink (or in the case of GTO it compresses) in diameter in order to stretch. The GTO works very much like this. When the collagen around the GTO is compressed because of contraction or stretch by the muscle, the Ib neurons generate an impulse that is proportional to the amount of GTO deformation. In this way the GTO can decrease contraction of a muscle being stretched in order to protect it from being torn. Likewise, GTO are thought to prevent unusually high contractions of a muscle in order to protect it from tearing itself apart. So in an antagonist muscle, the GTO can serve to inhibit co-contraction, facilitating contraction of the prime mover. In a prime mover, the GTO acts to prevent torn pecs, biceps and whatever else you are using to lift insanely heavy weights.

Another neuronal structure regulating involuntary muscle activity is the muscle spindle. The muscle spindle is found in greater abundance in the muscle belly as apposed to the musculotendonous junction. The muscle spindle also responds to stretch. However, the spindle is less like a Chinese finger trap and more like spring. When the muscle undergoes stretch, the center of the spindle is stretched. These spindles contain neurons that are sensitive to this stretching. Unlike with the GTO, when a muscle spindle is stretched its excitatory neurons fire in order to counteract the stretch. When a stretch is imposed on a muscle, the Type-I sensory neuron sends impulses into the spinal cord and connects with interneurons, generating an excitatory local-graded potential that is sent back to the muscle being stretched. If the stretch is of sufficient magnitude and/or rate, a local graded impulse will be sent back to the same muscle with sufficient strength to initiate a contraction via alpha motoneurons. This reflex arc in known as the "stretch-reflex" and is characterized by a quick muscular contraction following a rapid stretch of the same muscle. Now this stretch reflex primarily functions in slow twitch muscle fibers.

Alterations in the sensitivity of these two regulatory mechanisms have been seen with training. Carolan (Carolan, 1992) showed a decrease in antagonist co-activation of the lex extensors with training. On the other hand, increases in co-activation have been seen in longitudinal studies comparing explosive trained athletes to non-explosive trained athletes (Osternig 1986, Barrata 1988). These somewhat contradictory results may reflect the possibility that co-activation alterations are very specific in nature and depend on things such as contraction velocity, range of motion, and training specific effects.

Training for Size
Of primary interest to bodybuilders is training for size. After all, what is bodybuilding but doing whatever you can to make your muscles larger. The goal now is to use the current knowledge of the way muscle tissue reacts to imposed mechanical overload and microtrauma to plan a training strategy or routine that best elicits a growth effect. Understand that from here on out you are going to see areas where different approaches would be equally valid. One reason for this is the lack of quality research looking specifically at muscle hypertrophy in humans using typical (or atypical for that matter) exercise protocols that last for more than 8-12 weeks. In part 1 of this article we mentioned the fact that the length of a standard school semester or quarter usually dictate the length of a given study. Volunteers are hard to keep track of when their schedules change or when summer starts.

Conclusion
Thus far in Part II we have discussed the various mechanisms by which muscle adapts to resistance exercise. This is very important to understand if we are going to make advances in training techniques and planning. We know that in order for a muscle to undergo compensatory hypertrophy it must be subjected to sufficient trauma to activate satellite cells. This is done through the activity of various growth factors which all act in concert to regulate muscle hypertrophy. We also know that in order to increase our strength we must increase the "efficiency" of nerve conduction and motor unit coordination. Training to accomplish this relies heavily upon the principle of specificity which was spoken of in Part 1 of "Advanced Training Planning for Bodybuilders". If you simply want brute force you must train the nervous system and the resultant increase in strength will reflect the manner in which you trained. Now one should not assume that training for size will automatically lead to significant increases in size. This simply isn't true. In Part 3 we will examine specifically what methods can be adopted to cause increases in muscle size or increases in muscular strength. You might be surprised with what we come up with so stay tuned.

[einstein1905]

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Abraham, J. A., J. L. Whang, A. Tumolo, A. Mergia, J. Friedman, D. Gospodarowicz, and J. C. Fiddes. Human basic fibroblast growth factor: nucleotide sequence and genomic organization. EMBO J. 5: 2523-2528, 1986

Barrata R., Solomonow M., Zhou B., Letson D., Chouinard L., and D'Ambrosia R. Muscular co-activation: The role of the antagonist musculature in maintaining knee stability. Am. J. Sports Med. 16:113-122. 1988

Barrett E. J., Gelfand R. A. The in vivo study of cardiac and skeletal muscle protein turnover. Diabetes Metab. Rev. 1989; 5:133-148

Belanger AY., McComas AJ. Extent of motor unit activation during effort. J. Appl. Physiol. 51:1131-1135. 1981

Bellemare F., Woods JJ., Johansson R., Bigland-Ritchie B. Motor-unit discharge rates in maximal voluntary contractions of three human muscles. J. Neurophysiol. 50:1380-1392, 1983

Bennet, W. M., Connacher, A. A., Scrimgeour, C. M., Jung, R. T. and Rennie, M. J. (1990) Euglycemic hyperinsulinemia augments amino acid uptake by human leg tissues during hyperaminoacidemia. Am. J. Physiol. 259 (Endocrinol. Metab. 22): E185-E194.

Borer KT. The effects of exercise on growth. Sports Med 20(6):375-397 1995

Butchal F., Madsen T. Synchronous activity in normal and atrophic muscle. Electroencephal. Clin. Neurophysiol. 2:425-444. 1950

Carolan B., Cafarelli E. Adaptations in co-activation in response to isometric training. J. Appl. Physiol. 73:911-917. 1992

Chapman SJ., Edwards RH., Gregg C., Rutherford O. Practical application of the twitch interpolation technique for the study of voluntary contraction of the quadriceps muscle in man. J. Physiol. (Lond) 353:3P. 1985

Clamann HP., Schelhorn TB. Nonlinear force addition of newly recruited motor units in the cat hindlimb. Muscle and Nerve 11:1079-1089. 1988

Clarke MS, Feeback DL Mechanical load induces sarcoplasmic wounding and FGF release in differentiated human skeletal muscle cultures. FASEB J. 10(4):502-509 1996

Fryburg D. A. Insulin -like growth factor I exerts growth hormone - and insulin-like actions on human muscle protein metabolism. Am. J. Physiol. 1994; 267:E331-E336

Fryburg D. A., Barrett E. J. Growth hormone acutely stimulates skeletal muscle but not whole-body protein synthesis in humans. Metabolism 1993; 42:1223-1227

Fryburg D. A., Barret E. J., Louard R. J., Gelfand R. A. Effect of starvation on human muscle protein metabolism and its response to insulin. Am. J. Physiol. 1990; 259:E477-E482

Fryburg D. A., Gelfand R. A., Barrett E. J. Growth hormone acutely stimulates forearm muscle protein synthesis in normal human. Am. J. Physiol. 1991; 260:E499-E504

Gandevia SC., McKenzie DK. Activation of human muscles at short muscle lengths during maximal static efforts. J. Physiol. (Lond) 407:599-613. 1988

Gelfand R. A., Barrett E. J. Effect of physiologic hyperinsulinemia on skeletal muscle protein synthesis and breakdown in man. J. Clin. Invest. 1987; 80:1-6

Gual P, Baron V, Lequoy V, Van Obberghen E. Interaction of Janus kinases JAK-1 and JAK-2 with the insulin receptor and the insulin-like growth factor-1 receptor. Endocrinology 139(3):884-893, 1998

Hakkinen K, Komi PV. EMG changes during strength training and detraining. Med. Sci. Sports Exerc. 15(6):455-460. 1983

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Rosenblatt JD, Yong D, Parry DJ., Satellite cell activity is required for hypertrophy of overloaded adult rat muscle. Muscle Nerve 17:608-613, 1994

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[flexshack]

I'd assume they're referring to more long term. If the satellite cell population is depleted, even though this contributed to extensive regeneration of muscle, the causative agent still remains and confers disease, in the case of DMD, a genetic defect. So, inevitably the muscle that has been regenerated is still doomed to the same fate as the muscle it replaced.




Myogenic satellite cells: physiology to molecular biology
Thomas J. Hawke1 and Daniel J. Garry1,2
J Appl Physiol 91: 534-551, 2001;
8750-7587/01


"Most myopathies have a molecular mutation that affects the structural or cytoskeletal proteins in skeletal muscle. Duchenne muscular dystrophy (DMD) is the most common and the most devastating of the muscular dystrophies (20, 55, 76, 82, 127). Disease progression and death are ultimately due to a failure of the myogenic satellite cells to maintain muscle regeneration (36, 80). DMD is a recessive X-linked disease that results in a null mutation at the dystrophin locus (20, 82, 127). The absence of this cytoskeletal protein renders the muscle fiber extremely fragile. In response to mechanical stress associated with repeated contraction, there is widespread degeneration. The satellite cells respond to the injury by repopulating the injured skeletal muscle with defective myofibers lacking dystrophin. This process results in continuous degeneration-regeneration cycles and ultimately exhausts the satellite cell pool (36, 80).

Clinical symptoms are apparent by 4-5 yr of age in boys with DMD (10, 20, 127). DMD patients (4-5 yr of age) have been shown to undergo more skeletal muscle regeneration than that measured in a total of six normal patients over 60 yr of age (46). These results were confirmed by Renault et al. (151), who demonstrated that the proliferative life span of satellite cells derived from a 9-yr-old DMD patient was approximately one-third of an age-matched control. Proliferative fatigue or senescence of the satellite cell population and the milieu of the DMD skeletal muscle may collectively impair the proliferative or regenerative capacity of this cell population. In the DMD patient, increased levels of IGF binding proteins (IGFBP) are released by fibroblasts. The elevated IGFBP sequesters IGF-I, limiting its bioavailability for satellite cells and ultimately resulting in increased skeletal muscle fibrosis (120). The evidence to date demonstrates the tremendous strain and ultimate failure of the satellite cell population to adequately compensate for the persistent degeneration-regeneration process that is occurring in the DMD skeletal muscle."

wow, nice digging/researching. thanks. i'm always impressed einstein.

[flexshack]

Here is another study showing testosterone having a role in satellite cell activathion. However, the study is generalized and doesn't look at specifics. We know that testosterone increases GH, which increases IGF-1, so really all this study says to me is .....what I just said, but AR has lots of bandwidth, so I'm posting it anyway.

Dev Biol. 1995 May;169(1):286-94. Related Articles, Links


Testosterone treatment results in quiescent satellite cells being activated and recruited into cell cycle in rat levator ani muscle.

Joubert Y, Tobin C.

Laboratoire de Neurobiologie des signaux intercellulaires, URA 1488, Paris.

At puberty, the male levator ani (LA) muscle exhibits muscle fiber hypertrophy. This fiber enlargement can be provoked in the female LA muscle by testosterone treatment. In both cases the hypertrophic process is accompanied by an increase in the number of satellite cells and myonuclei. The present ultrastructural autoradiographic study was undertaken in order to investigate (1) whether satellite cells when stimulated by testosterone can undergo DNA synthesis by incorporating [3H]thymidine ([3H]Tdr); (2) whether the whole satellite cell population is committed in the cell cycle; and (3) whether the new myonuclei originate from fusion of the satellite cells with the preexistant myofibers. Thirty-day-old female rats treated with a single testosterone injection received a single injection of [3H]Tdr at 24, 28, 32, to 34 hr after testosterone treatment. LA muscles were removed 2 hr after [3H]Tdr injection. This first series of experiments allowed us to determine that onset of DNA synthesis in satellite cells occurs within the 34th and the 36th hour after testosterone treatment. To obtain a more precise timing, 30-day-old female rats treated with a single testosterone injection received multiple injections every 30 min either from Hour 32 to 33.5 or from Hour 34 to 35.5 LA muscles were removed 30 min after the last [3H]Tdr injection. This showed that the onset of satellite cell replication occurred between the 32nd and the 34th hour after testosterone treatment. However, only 30% of the satellite cell population was affected by this proliferative process regardless of the experimental protocol used. To confirm that the increase in myonuclei number results from incorporation of satellite cells into mature myofibers, 30-day-old female rats treated with a single injection of testosterone received a single injection of [3H]Tdr on the 60th hour after testosterone treatment. LA muscles were removed at 63, 84, to 108 hr after [3H]Tdr injection. We conclude that testosterone induces satellite cell proliferation at around the 33rd hour and that the increased number of myonuclei reported in our previous study is due to fusion of labeled satellite cells with myofibers.


Here is the Figure legend for the attached image:
Fig. 4. Satellite cell response to myotrauma. *Skeletal muscle trauma or injury may be minor (e.g., resistance training) or may be more extensive (e.g., toxin injection, Duchenne muscular dystrophy). In response to an injury, satellite cells become activated and proliferate. Some of the satellite cells will reestablish a quiescent satellite cell pool through a process of self-renewal. Satellite cells will migrate to the damaged region and, depending on the severity of the injury, fuse to the existing myofiber or align and fuse to produce a new myofiber. In the regenerated myofiber, the newly fused satellite cell nuclei will initially be centralized but will later migrate to assume a more peripheral location.

when you said testosterone increases gh and therefore igf-1, is it primarily due to the converted estrogens that cause this increase or does testosterone itself have this ability? i know estrogen causes significant elevation of igf-1 levels, but what about androgens (maybe to a lesser degree)?

[einstein1905]

when you said testosterone increases gh and therefore igf-1, is it primarily due to the converted estrogens that cause this increase or does testosterone itself have this ability? i know estrogen causes significant elevation of igf-1 levels, but what about androgens (maybe to a lesser degree)?

Estrogen does play a significant role. However, most clinical studies that have included both women and men receiving GH treatment have shown women respond less favorably, especially in regards to strength/mass increases.

[NewBreed]

Women may have a weaker GH-receptor-response,since high IGF-1-levels are not favorable in women,because it promotes breast-cancer.
A Havard-studie showed,that even milk elevates IGF in women thus leading to a significant increase in breast cancer trials,also negotiating poitive effects of multile births on breast cancer.

[flexshack]

Advanced Training Planning for Bodybuilders: Part 2
written by Bryan Haycock


Introduction
In part one of this article we discussed the history of resistance training and some very basic principles at work during adaptation to resistance training. Specificity was outlined as a governing principle with which we can predict the outcomes of our training. Low volume/high load training produce increases in neuromuscular efficiency and motor unit recruitment, while high volume/moderate load training produces only moderate increases in strength and neuromuscular adaptations along with marked hypertrophy of both slow and fast twitch fibers. Also discussed were issues such as rational and irrational adaptation. Dramatic increases in sarcomere volume without increases in myo-nuclear number, seen during irrational adaptation, effectively inhibits further increases in the production of contractile proteins and diminishes recovery and performance. Slower increases in sarcomere volume, as seen in rational adaptation, actually facilitates recovery and leads to a more steady increase in both size and strength. In part two we will discuss the mechanisms responsible for the specific nature of adaptation and look at ways of applying this knowledge to build size and strength.

The Stimulus for Muscle Growth
To most people, the way to get a muscle to grow larger is simply a matter of "exercising" the muscle you want to grow. This is a very simplistic way of looking at it. Those individuals who fail to realize that there is a right way and a wrong way to train a muscle never go on to develop out of the ordinary physiques. Don't get me wrong, the knowledge of how muscle tissue grows in response to training is not a requirement to be a successful bodybuilder. Anabolics have become the ultimate "cheat sheet". They effectively reduce the overload threshold necessary for compensatory hypertrophy and elevate the genetic limit or plateau. If you chose not to use potent synthetic hormones you are in for a much more difficult road, and a good understanding of muscle hypertrophy will be invaluable to you as you train over the years.

At the very foundation of muscle hypertrophy is load induced tissue strain. Muscle tissue must undergo mechanical strain in order to begin the biochemical steps necessary for adaptive growth (Clarke,1996). This "strain" induced by the loading of the tissue leads to sarcolemma microtrauma. The wounding of the muscle cell after training is characterized by myofibrillar disruption, Z line smearing, discontinuity of sarcomeres and an increase in the porosity, or permeability, of the sarcolemma. The effect or result of this nonfatal cellular damage is the production, and release of growth factors that then interact with the damaged cell itself and also very importantly, with satellite cells.

Satellite cells are myogenic stem cells that serve to assist postnatal growth and regeneration in adult skeletal muscle. Following proliferation (reproduction) and subsequent differentiation (to become a specific type of cell), these satellite cells will fuse with one another or with the adjacent damaged muscle fiber, thereby increasing myonuclei numbers for fiber growth and repair. Proliferation is necessary in order to meet the needs of thousands of muscle cells all potentially requiring additional nuclei. Differentiation is necessary in order for the new nucleus to behave as a nucleus of muscle origin. In order to better understand what is physically happening between satellite cells and muscle cells, try to picture 2 oil droplets floating on water. The two droplets represent a muscle cell and a satellite cell. Because the lipid bilayer of cells are hydrophobic just like common oil droplets, when brought into proximity to one another in an aqueous environment, they will come into contact for a moment and then fuse together to form one larger oil droplet. Now whatever (i.e. nuclei) was dissolved within one droplet will then mix with the contents of the other droplet. This is a simplified model of how satellite cells donate nuclei to existing muscle cells.

There appears to be a finite limit placed on the cytoplasmic/nuclear ratio (Rosenblatt,1994). Whenever a muscle grows in response to functional overload there is a positive correlation between the increase in the number of myonuclei and the increase in fiber cross sectional area (CSA). When satellite cells are prohibited from donating viable nuclei, overloaded muscle will not grow (Rosenblatt,1992; Phelan,1997). It is not a stretch to say that satellite cell activity is a required step, or prerequisite, in compensatory muscle hypertrophy, for without it, a muscle simply cannot significantly increase total protein content nor CSA.

Some factors which regulate this process are exercise, trauma, passive stretch, massage, innervation, and the activity of soluble growth factors. Three classes of growth factors in particular have been studied extensively with respect to their effects on satellite cell proliferation and differentiation in vitro. They are; fibroblast growth factors (FGF), insulin -like growth factors (IGF), and the transforming growth factor-beta superfamily (TGF-beta). When administered in combination, these factors can induce satellite cell activities in vitro which mimic those typical of satellite cells found in vivo in growing, regenerating, or healthy mature muscle. In essence they can mimic the effects of loading without any microtrauma actually being done.

FGF is one growth factor being actively studied. Recent studies have shown that an increase in the permeability of the sarcolemma is necessary for the release of fibroblast growth factor (FGF). This is because FGF does not contain a signal transduction peptide sequence (Abraham,1986) and thus does not exit the source cell through vesicle mediated exocytosis. This "signal transduction peptide sequence" is necessary for the protein to be incorporated into intracellular vesicles and actively transported to the cell surface, and then ultimately released. Instead, it must be able to pass directly through the phospholipid bilayer of the wounded sarcomere in order to have an autocrine and paracrine effect on target tissues. This is accomplished in adult muscle fibers by inflicting microtrauma.

FGF, specifically FGF-beta, has been shown to stimulate proliferation but suppress differentiation of myogenic stem cells. This has the result of increasing the availability of satellite cells but reduces the number of them that are actively fusing with nearby muscle cells. In a study where FGF was injected directly into healthy muscle tissue, it had the effect of increasing muscle DNA content and IGF-1 peptides, but had no significant effect on total protein content or gross muscle weight (Adams,1998). These researchers speculate that the increase in DNA content was the result of increased satellite cell number. The fact that FGF was unable to induce hypertrophy reflects the fact that FGF inhibits satellite cell differentiation (i.e. the satellite cells never actually became of the muscle cell type) preventing the nuclei from producing muscle specific proteins and thereby short circuiting a critical step the growth process. An additional study involving both local injection and local implantation of FGF pellets into "regenerating", or damaged muscle, failed to show any effect of FGF on the ability of muscle tissue to regenerate after microtrauma (Mitchell,1996). In this study FGF did have the effect of enhancing satellite cell proliferation as well as angiogenesis (capillary formation) nevertheless, it appears that FGF is not the limiting factor in compensatory muscle hypertrophy.

Insulin-like growth factor (specifically IGF-1) stimulates both proliferation and differentiation in an autocrine-paracrine manner, although it induces differentiation to a much greater degree. IGF-1, when injected locally, increases satellite cell activity, muscle DNA, muscle protein content, muscle weight and muscle cross sectional area (Adams,1998). As discussed earlier, the proliferation and differentiation of satellite cells is critical part of compensatory hypertrophy. The importance of IGF-1 lies in the fact that all of its apparent functions act to induce muscle growth with or without overload although it really shines as a growth promoter when combined with physical loading of the muscle.

IGF-1 also acts as an endocrine growth factor having an anabolic effect on distant tissues once released into the blood stream by the liver. In human volunteers, detailed information on the effect of IGF-1 on protein synthesis, degradation and balance has been obtained by using the arteriovenous difference of labeled and unlabeled phenylalanine across the forearm (Barrett and Gelfand 1989). In the aforementioned studies, "systemic" IGF-1 infusion for 6 h caused positive amino acid balance, both by inhibiting protein degradation and stimulating protein synthesis (Fryburg, 1994). This differs from the effect of peptide "hormones" such as insulin, which does not stimulate synthesis in adults (Bennet et al. 1990, Fryburg 1990, Gelfand and Barrett 1987, McNurlan, 1994). Therefore IGF-1 possesses the insulin-like property of inhibiting degradation, but in addition can stimulate protein synthesis (Fryburg 1991). The insulin-like effects are probably due to the similarity of the signaling pathways between insulin and IGF-1 following ligand binding at the receptors (Schumacher 1991, Gual 1998).

The ability of IGF-I to stimulate protein synthesis resembles the action of GH, which was shown in separate studies on volunteers to stimulate protein synthesis without affecting protein degradation (Fryburg et al. 1991, Fryburg and Barrett 1993). Although it is often believed that the effects of GH are mediated through IGF-1, this cannot be the case entirely. First, the effects of the two hormones were different, in that GH did not change protein degradation. Second, the effect of GH was observed with little or no change in systemic IGF-1 and GH concentrations because the GH was infused directly into the brachial artery (Fryburg et al. 1991).

Transforming growth factor-beta (TGF-beta) slightly depresses proliferation but inhibits differentiation. So although it is called a growth factor, it is an "inhibitory" factor involved in muscle growth. For example, one TGF-beta member known as growth/differentiation factor-8 (GDF-8), was found to have profound effects on muscle growth (McPherron and Lawler,1997). GDF-8 was found to exist in many muscles throughout the body. In order to identify the function of this protein, researchers "targeted", or "knocked out" the gene responsible for producing it. The result was nothing less than miraculous. The mice who lacked the gene went on to grow muscles up to 3 times larger than normal mice. This is a 300% increase in muscle weight with no specific exercise or mechanical loading! You may remember pictures of these mice published in issue number 188 in MuscleMag International (MuscleMag,1998). The increase in the mass of the "mutant" mice muscles was a result of both increased hypertrophy and hyperplasia (an increase in fiber number). There are also naturally occurring mutations to this gene that result in "double muscling" of animals such as livestock. Two breeds of cattle known as Belgian Blue and Piedmontese exhibit naturally occurring mutations on the myostatin gene (McPherron and Lee,1997). This changes the amino acid sequence of the GDF-8 peptide and greatly attenuates its physiological activity.

All of these growth factors are brought into play in human models of compensatory hypertrophy. In summary, resistance exercise of sufficient load and volume causes microtrauma to the sarcolemma which increases the release and production of growth factors. This is done through increased permeability of the wounded cell membrane allowing soluble growth factors to "leak" out into the intercellular space. The growth factors then go on to increase the number, or "proliferation" of satellite cells, also called myogenic stem cells. This is done through interaction with growth factor receptors on the surface of these cells. These growth factors then go further to induce the conversion, or "differentiation" of these undifferentiated cells into cells expressing DNA of muscle origin. Once these satellite cells have undergone differentiation they can then fuse to existing muscle cells. This fusion allows the satellite cell to donate needed myonuclei to the wounded or developing muscle cell. The effect of increasing the number of nuclei within a cell allows for increased protein synthesis and ultimately hypertrophy. As mentioned earlier, there is a nuclear to cytoplasmic ratio, or "nuclear domain", that is closely regulated by the cell. If the muscle cell is prevented from increasing the number of nuclei, it does not grow in response to overload (Phelan,1997).

All of the aforementioned processes occur with or without systemic hormonal influence or nutritional abundance (Borer,1995). In this way the hypertrophic response can be limited to the overloaded tissue. I do not mean to imply that exercise induced growth is not effected by systemic hormones and/or nutritional abundance, only that mechanisms are in place that allow for growth in localized muscle tissue in the absence of endocrine support and adequate nutritional status.

The Stimulus for Strength
The foundation for the development of strength is neuromuscular in nature. Increases in strength from resistance exercise has been attributed to several neural adaptations including altered recruitment patterns, rate coding, motor unit synchronization, reflex potentiation, prime mover antagonist activity, and prime mover agonist activity. Aside from incremental changes in the number of contractile filaments, voluntary force production is largely a matter of "activating" motor units. In order to ascertain the relative contribution of each of these mechanisms, various measurement techniques have been utilized. Hereafter we will briefly discuss each of these mechanisms as they relate to resistance training.

Recruitment of motor units can be measured with Electromyography (EMG). As a muscle contracts, the electrical signal initiated by the motor nerve can be detected with EMG. The intensity or magnitude of this signal is sometimes described as "neural drive". In order to explain increases in strength from resistance exercise, researchers have measured the changes in EMG activity in weight training subjects.

Hakkinen and co-workers have shown that there is an increase in EMG activity with strength training as well as a decrease in EMG activity upon cessation of training (Hakkinen,1983). Fourteen male subjects (20-30 yr) accustomed to weight training went through progressive strength training of combined concentric and eccentric contractions three times per week for 16 wk. The active training period was followed by an eight week detraining period. The training program consisted mainly of dynamic exercises for leg extensors with the loads of 80-120% of one maximum concentric repetition (1RM). Significant improvements in muscle function were observed in early conditioning; however, the increase in maximal force during the very late training period was greatly limited. Marked improvements in muscle strength were accompanied by significant increases in the neural activation (EMG) of the leg extensor muscles. The relationship between EMG and high absolute forces changed during the training period. The occurrence of these changes varied during the course of training. During detraining, there was a decline in EMG activity.

Now those who would argue that increases in strength are solely due to increased recruitment of motor units would have a difficult time defending themselves in light of other research. The is a method of measuring motor unit activity called "Interpolated Twitch Technique", or ITT. ITT is used to determine the extent of activation of the entire muscle. Merton (Merton, 1954) was the first to use this technique to describe whole muscle activation. He showed full activation of the adductor pollicis with fatigue in untrained subjects. Several other studies have since shown a similar ability of untrained subjects to voluntarily fully activate various muscle groups (Bellemare 1983, Chapman 1985, Gandevia 1988, Belanger 1981). This directly contradicts the theory of strength increases due to the ability to activate more motor units.

The activation of motor units is done in an asynchronous fashion, meaning that not all fibers contract at the same time within a given muscle. There is a hierarchy to the order of fiber recruitment in muscle tissue. Because fiber activation is not "analog" or variable in nature, in other words, a fiber is either fully activated or fully quiescent, the brain must control contraction intensity by altering the number of fibers it activates. In general, slow twitch fibers are activated first followed by larger fast twitch fibers. Now when muscles begin to fatigue the asynchronous firing of fibers become more and more synchronized (Butchal, 1950). This allows for greater force production. This synchronization of muscle fibers has been linked to increases in voluntary strength (Milner-Brown, 1975). Now although increases in motor unit synchronization have been reported with training, studies involving artificial stimulation show that force development with asynchronous stimulation is greater and smoother (Clamann, 1988). In addition, researchers have shown that the rate of force development in brief maximal contractions is faster in voluntary than in evoked contractions (Miller, 1981). So from these studies we see that although synchronization of motor units can increase with training, asynchronous motor unit activation is more advantageous to rate and magnitude of force development than is synchronous activation.

Increases in "reflex potentiation" have also been linked to resistance training (Sale and Upton 1983, Sale and MacDougall 1983) as well as decreases with immobilization (Sale, 1982). The actual benefit, if any, of this adaptation is unclear. An increase in reflex potentiation would contribute to the voluntary EMG signal augmenting the motor or neuronal drive. Nevertheless, because untrained individuals have been shown to be able to fully recruit their motor units, the purpose of increased reflex potential remains undecided.

Finally, that activity of prime mover agonists and antagonists plays a role in directed voluntary strength. The obvious role of agonists is to assist the prime mover by guidance and stabilization. This could be termed "coordination". It is well known that any unaccustomed exercise requires practice in order to develop sufficient coordination to allow maximum efficiency of muscular effort. The role of antagonistic muscle groups is more complicated. They serve to prevent damage through co-contraction as well as ensure less resistance through relaxation to prime mover contractility.

The protective mechanisms function by way of golgi tendon organs (GTO). The GTO is sensitive to force output or tension within the muscle. They are located at the musculo-tendonous junction and is contained within a compressible collagenous capsule. Fibers of the GTO are connected directly to muscle fibers as well as to Type "Ib" inhibitory neurons within the muscle. The physical structure of the GTO allows it to be sensitive to stretch or load present in the muscle. Think of the notorious "Chinese finger trap". You first stick you fingers in each end. Then as you pull your fingers apart, the structure of the woven tube causes it shrink (or in the case of GTO it compresses) in diameter in order to stretch. The GTO works very much like this. When the collagen around the GTO is compressed because of contraction or stretch by the muscle, the Ib neurons generate an impulse that is proportional to the amount of GTO deformation. In this way the GTO can decrease contraction of a muscle being stretched in order to protect it from being torn. Likewise, GTO are thought to prevent unusually high contractions of a muscle in order to protect it from tearing itself apart. So in an antagonist muscle, the GTO can serve to inhibit co-contraction, facilitating contraction of the prime mover. In a prime mover, the GTO acts to prevent torn pecs, biceps and whatever else you are using to lift insanely heavy weights.

Another neuronal structure regulating involuntary muscle activity is the muscle spindle. The muscle spindle is found in greater abundance in the muscle belly as apposed to the musculotendonous junction. The muscle spindle also responds to stretch. However, the spindle is less like a Chinese finger trap and more like spring. When the muscle undergoes stretch, the center of the spindle is stretched. These spindles contain neurons that are sensitive to this stretching. Unlike with the GTO, when a muscle spindle is stretched its excitatory neurons fire in order to counteract the stretch. When a stretch is imposed on a muscle, the Type-I sensory neuron sends impulses into the spinal cord and connects with interneurons, generating an excitatory local-graded potential that is sent back to the muscle being stretched. If the stretch is of sufficient magnitude and/or rate, a local graded impulse will be sent back to the same muscle with sufficient strength to initiate a contraction via alpha motoneurons. This reflex arc in known as the "stretch-reflex" and is characterized by a quick muscular contraction following a rapid stretch of the same muscle. Now this stretch reflex primarily functions in slow twitch muscle fibers.

Alterations in the sensitivity of these two regulatory mechanisms have been seen with training. Carolan (Carolan, 1992) showed a decrease in antagonist co-activation of the lex extensors with training. On the other hand, increases in co-activation have been seen in longitudinal studies comparing explosive trained athletes to non-explosive trained athletes (Osternig 1986, Barrata 1988). These somewhat contradictory results may reflect the possibility that co-activation alterations are very specific in nature and depend on things such as contraction velocity, range of motion, and training specific effects.

Training for Size
Of primary interest to bodybuilders is training for size. After all, what is bodybuilding but doing whatever you can to make your muscles larger. The goal now is to use the current knowledge of the way muscle tissue reacts to imposed mechanical overload and microtrauma to plan a training strategy or routine that best elicits a growth effect. Understand that from here on out you are going to see areas where different approaches would be equally valid. One reason for this is the lack of quality research looking specifically at muscle hypertrophy in humans using typical (or atypical for that matter) exercise protocols that last for more than 8-12 weeks. In part 1 of this article we mentioned the fact that the length of a standard school semester or quarter usually dictate the length of a given study. Volunteers are hard to keep track of when their schedules change or when summer starts.

Conclusion
Thus far in Part II we have discussed the various mechanisms by which muscle adapts to resistance exercise. This is very important to understand if we are going to make advances in training techniques and planning. We know that in order for a muscle to undergo compensatory hypertrophy it must be subjected to sufficient trauma to activate satellite cells. This is done through the activity of various growth factors which all act in concert to regulate muscle hypertrophy. We also know that in order to increase our strength we must increase the "efficiency" of nerve conduction and motor unit coordination. Training to accomplish this relies heavily upon the principle of specificity which was spoken of in Part 1 of "Advanced Training Planning for Bodybuilders". If you simply want brute force you must train the nervous system and the resultant increase in strength will reflect the manner in which you trained. Now one should not assume that training for size will automatically lead to significant increases in size. This simply isn't true. In Part 3 we will examine specifically what methods can be adopted to cause increases in muscle size or increases in muscular strength. You might be surprised with what we come up with so stay tuned.

well i finally got to this one and it was an excellent read. i definately learned a lot.

einstein, i was wondering, with the body's regulatory or homeostatic tendencies, is there a possibility that it would compensate for supraphysiological levels of igf-1 by releasing more levels of TGF-beta or other "inhibitory growth factors"?

also, do you have part 3 to this b/c i'd love to read it?

again, excellent post.