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marylin monroe
Showing posts with label signaling. Show all posts
Showing posts with label signaling. Show all posts

Beyond Satellite Cells: Eccentric Training Bullies Bystanding Stem Cells, Destined to Become Bone, Cartilage, Adipose or Nerve Tissue into Contributing to Skeletal Muscle Growth

Image 1: Apart from their epistemic value, these stained tissue samples from the Valero study are actually pretty aesthetic.The arrows, by the way, mark NG2 monocytes, which "coerce" the non-myogenic stem cells (perycites) to get going ;-)
I guess, the term "satellite cells" has been mentioned so often in the course of the past couple of weeks that it actually would not be necessary to tell you (once again) that these myogenic precursor cells are a necessary prerequisite for the repair and long-term growth of your pecs, legs, back, biceps, triceps, delts and the rest of the skeletal muscle tissue in your body. Those of you, who have read all the latest installments of the Intermittent Thougths will also be familiar with the notion that both the local expression of MGF-1 and inflammatory cytokines, as well as systemic hormones, such as testosterone and estrogen play an important role not only in the recruitement and migration of satellite cells into the muscle, but also in their regeneration and maintenance (cf. "Are You Serming Away Your Growth Potential"). A group of scientists from the University of Illinois must however have figured that this is not yet complicated enough and began digging even deeper into the (re-)generation of (new) muscle tissue... and let me tell you, what they found is intriguing.

Eccentric training is "numero uno" for satellite cell recruitment

I guess you will be aware that it is a relatively well-established fact that of all real-world physical activities eccentric, or lengthening, contractions of skeletal muscle appears to constitute the most potent inducer of both "productive" exercise induced muscle damage, as well as subsequent increases in satellite cell recruitment.
Figure 1: Changes in intramuscular architecture in young and old subjects in response to 6 sets of eccentric leg extensions (data calculated based on Dreyer. 2006)
Back in 2006 Dreyer et al. published the results of a study which compared the satellite cell response to an eccentric exercise regimen (1x12 + 5x16 eccentric-only reps on leg-extension machine) in young and old subjects (Dreyer. 2006). As the data in figure 1 goes to show, this unquestionably exhaustive bout of exercise produced a quite remarkable (>150%) increase in satellite cell volume per muscle fiber and a likewise highly significant (>100%) increase in the number of satellite cells compared to the total number of cells in the sublaminar compartment. It is also evident from the data in figure 1 that the preparatory accumulation of satellite cells 24h after the eccentric exercise-bout was profoundly blunted in the older subjects.
Figure 2: Satellite cell count per myonucleus before and 8d after 100reps of eccentric leg extensions in untrained young subjects (data adapted from Mikkelson. 2009)
Note: As you can see in figure 2, age is yet not the only factor that can compromise the adaptive response to eccentric lengthening contraction of skeletal muscle. Mikkelson et al. who had the (young) participants of their study perform 100 eccentric reps on a similar leg-extension machine (Mikkelson. 2009), for example, found that an indomethacin (NSAID; COX-inhibitor) infusion for 7.5 h during the exercise day did not only blunt the increase in pax-7+/myonuclei, or in "layman's terms", satellite cells 8 days after the exercise session, the combination of exercise and NSAID actually led to a slight and statistically obviously non-significant reduction in satellite cells per myonucleus. This does support the findings, I discussed in one of the previous installments of the Intermittent Thoughts, which dealt with the importance of "inflammation" as a vital constituent of both the repair and hypertrophy response to exercise induced muscle-damage (cf. "IGF-1, IL-15 & Co").
The age-related decline in satellite cell activity unquestionably raises a question Lerner addressed in one of his insightful comments on my post on the important role of estrogen for the maintenance of the satellite cell pool, which is whether or not the latter would be limited... I mean, when the whole craze about stem-cell therapy began, the general accepted notion appeared to be that the average adult human being has a very limited / if any of those "super cells".

If you have been following recent publications, you will yet be aware that as of late researchers (interestingly also from the University, I work at ;-) have made quite some progress in "producing" and "reprogramming" stem cells from all sorts of human and animal tissue. Assuming that you have also read everything about how testosterone works its "muscle building and fat burning magic" (cf. "Understanding the Big T"), you should also be aware that the latter, i.e. the "fat burning" effect is at least partly mediated by the reprogramming of stem cells which are actually supposed to become fat cells into "satellite cells". All that being said, it should actually not surprise you that the main finding of the initially mentioned study is that eccentric contractions of skeletal muscle have a very similar effect on "non-myogenic" stem cells, which "happen to be in the vicinity of the exercised muscle fibers".

A study on You, Wolverine and the Ultimate Hard Gainer

Unfortunately, the design of the study by M. Carmen Valero and her colleagues from the Department of Kinesiology and Community Health at the University of Illinois is... well, let's say not exactly straight-forward, or easy to understand (Valero. 2012). Basically, the scientists took 3 types of muscle fibers from rodents,
Figure 3: Localization of stem cell antigen-1 (Sca-1) (arrows, TRITC-red) positive mononuclear cells and a-7 integrin (FITC-green) in the different muscle tissue before (SED) or 24 hr post-exercise (Ex) at 20x magnification (adapted from Valero. 2012)
  • wild type, as a normal control (that would be your muscle ;-)
     
  • alpha-7 integrin transgenic (a7TG), which is resistant to injury, but still responsive to strain (I guess that is either the "Wolverine" or "Unbreakable" type of muscle tissue ;-), and
     
  • alpha-7 (-/-), which is the "ultimate hard gainer muscle" that does not respond with alpha-7 integrin expression to overload
Now, even if you don't understand a word of what I am talking about here, I guess that you will be able to see the significant color-differences in the immuno-stained (=marked by antibodies) tissue samples in figure 3. If you just focus on the arrows, the red and green areas, that should suffice to grasp the idea that the both the stretch induced alpha-7 integrin response, as well as the number (arrows) and area (red staining) of stem cells in are maximal in the Wolverines, "normal" in guys and gals like you and minimal to non-existant in the "ultimate hard gainer".

The first take-home message of this study is thusly that the strain that is induced by eccentric training activates "dormant" stem cells via alpha-7 integrin. Why this is the case, becomes clear when you look at the structure of those heterodimers, which transverse the cell membranes of regular muscles and adhere the extracellular matrix to the cytoskeletal network. If you strain the muscle, this will obviously affect the integrin system, which thusly functions as a "sensor" for mechanical signals.

"No satellite cell available? Well I guess we just take this one, then..."

Upon closer analysis of the "satellite cells" that actually responded to the alpha-7 integrin signals, the scientists did yet realize that the resource from which the majority of the new muscle cells were recruited were actually mesenchymal stem cells, which are usually destined to become osteoblasts (bone), chrondocytes (cartilage) and adipocytes (fat cells):
In this study, we provide the first demonstration that muscle resident mesenchymal-like stem cells (mMSCs), predominantly pericytes, are increased in muscle in an a-7 integrin dependent
manner following an acute bout of eccentric exercise. mMSCs maximally appeared in a7BX2 transgenic muscle resistant to injury following eccentric exercise and were rarely present in
muscle lacking the a-7 integrin, suggesting that factors other than injury or inflammation are primary regulators of mMSC accumulation in skeletal muscle.
In view of the fact that a preliminary experiment in which the scientists transplanted exogenous mMSCs into the muscle of living mice, led to a very localized, but distinctly measurable increase in new fibers opens a whole new venue for research into the artificial enhancement of skeletal muscle hypertrophy... for you as an avid (and probably "natural") trainee, however, the main take home message is that exercise, in this case, the strain (not the damage!) that is induced by eccentric lengthening contractions of your muscles still appears to be the major determinant of all aspects of skeletal muscle hypertrophy. Don't forget that, when you are about to invest the next few hundred bucks into whatever "myogenic" supplement the guy at GNC is trying to persuade you into buying... without hard and consistent work at the gym those powders and caps are pretty useless.

    Chronic Resistance Training Reduces the Anabolic Signaling in Response to Exercise - 12 Days of Detraining Restore It

    This rodent obviously knows about the value of detraining as a means to restore the signaling protein response that gets blunted over weeks of continuous training (photo from livescience.com)
    Ah, some really good stuff in the news, or rather in the journals as of late (for the news version of the articles, you obviously got to come here, to the SuppVersity ;-). So, let's skip any lengthy preludes and let's start with a simple question pertaining to the topic of the day: "When was the last time you took 2 weeks or more off?" What? Last year, when you were down with the flu? No, that does not count. I am talking about detraining, here; so only voluntary off-times will be reckoned as off time... I thought so, you haven't taken off in years, right? Well, what if I told you that this may be the reason your gains have not taken off either? Interested? Yeah, that's what I thought.

    Chronic resistance training reduces its own anabolic effect, detraining restores it

    I guess in the end, all of us knew this instinctively: The unbelievable gains you make as a rookie vs. the slow and arduous road you will be walking later in your "career" as a trainee are too obvious for anyone not to suspect that the marginal utility of exercise declines.

    Learn more about domain sizes, protein synthesis and  "muscle bulding" in Part II of the Intermittent Thoughts on Building muscle.
    Now, one of the most common and certainly accurate hypothesis to explain this phenomenon is that the restructuring processes that starts when you hit the early domain size limit is more time consuming than just "pumping more protein into the muscle" - a process, which happens more or less automatically, a previously sedentary individual picks up a dumb- or barbell ;-)

    With the impending publication of a study by researchers from the Ritsumeikan University, the University of Tokyo, the Nippon Sport Science University and the  University of Mississippi we do now have evidence for another, yet probably not unrelated reason to the exponential decline in marginal utility: The amelioration of the exercise induced phosphorylation of signaling proteins, due to which the marginal utility of your workouts decreases over time.

    What did the scientists do?

    In the experiment, the results of which Riki Ogasawara and his colleagues summarized an discussed in their latest paper, the researchers randomized a group of male Sprague-Dawley rats (10 weeks of age, 356.1 ± 4.4 g body weight) to four groups (+control) performing either continuous training (1S, 12S, 18S), in the form of 1 exercise session (1S), 12 exercise sessions (12S), or 18 exercise sessions (18S) every other day (Mo, Tue, Wed, Fr, Su, Tue, ...), or continuous training + detraining (DT), in the form of 12 sessions of exercise every followed by 12 days of detraining.
      Figure 1: The "muscle builder" mTOR and the pertaining signaling cascade(s); remember that the line with the bar at the end indicates an antagonism → higher mTOR = higher phosphorylation of p70SK and it's downstream target rpS6, but lower 4E-BP1 (my orange markups; original from ebiotrade.com).
      The exercise itself was mimicking a leg-training regimen, in the course of which, the gastrocnemius muscle was trained by stimulating 5 contractions, with a 5-s interval between contractions, per set for 5 sets (5-min rest intervals in-between the sets).
      "The voltage (~30 V) and stimulation frequency (60 Hz) were adjusted to produce maximal isometric tension. Before every exercise session, peak twitch torque was measured. Torque signals were collected continuously at a sampling rate of 1024 Hz using a 16-bit analog-to-digital converter (PowerLab/16SP;AD Instruments, Japan) and analyzed using Power Lab Chart 5 software (AD Instruments, Japan). " (Ogasawara. 2013)
      24h after the last exercise session, the rats were anesthetized and exsanguinated. The muscles were removed immediately after death and both muscle size, volume and weight, as well as the expression of the signaling proteins p70S6 kinase, p90RSK, 4E-BP1 and S6 ribosomal protein (rpS6) were measured.

      So what's that all about? Did the rats become more muscular?

      As you can see from  my plot of the protein responses on the left and the respective effects the different training (+detraining) protocols had on the muscle weight of the rodents on the right hand side of figure 2, the chronic resistance training protocols lead to statistically significant reductions in the post training p70S6K and rpS6 expression, which were restored in response to the detraining protocol.
      Figure 2: Phosphorylation status of p70S6K (Thr389), p90RSK (Thr573), 4E-BP1 (total) an rpS6(Ser235/236) on the left and body weight, as well as muscle weight  in both exercised (RT ) and non-exercised (CON) rodents measured on the day after the last workout of the respective training group (Ogasawara. 2013).
      Notwithstanding, the fact that the muscle gains were (expressed relative to the respective control group) statistically identical, shouldn't surprise you. After all, the growth benefits of the detraining protocol will show only in the weeks after your absence from the gym. The rodents in the study at hand, however, were subjected to only one training session after the detraining period and killed afterwards. Therefore, the main message the data on the increase in muscle weight in the DT compared to its control group is sending us is that short periods of detraining won't cost you precious muscle mass.

      Don't hesitate, dare growing like a rookie again - dare taking a week off!

      In a previous study, by Ogasawara et al. I have likewise discussed here at the SuppVersity, the researchers have already shown that a "6-weeks-on vs. 3-weeks off" training-detraining regimen produces identical gains in muscle growth as continuous training w/out  producing the logarithmic decline in marginal gains that brings skeletal muscle hypertrophy to a screetching halt over time (read more)
      The novel information about the decline and restoration of the signaling proteins this study provides would support the longstanding hypothesis that continuous exercise blunts its own growth response. And what's more it's also supported by various human studies. In 2006, for example, Coffey et al. were able to show that the phosphorylation of p70S6K and rpS6 in response to resistance training was almost completely blunted in highly resistance-trained subjects (power lifters), while it did occur in the untrained controls (Coffey. 2006).Morever, Ogasawara et al. have shown last year already that an even longer detraining period of 3 weeks lead to much steeper inclines in muscle CSA in human subjects than continous training (see figure next to the paragraph below and read up all the details in the respective SuppVersity article from October 2012)

      Collectively, these results clearly suggest that the notion of planned, regular detraining periods could have benefits that go way beyond the well-known ability to protect you from getting caught in the downward spiral of chronic overtraining, as it will also "reset" the anabolic response to a given workload.

      And while you can hardly expect the results to be anywhere similar to those you've hopefully experienced, when you were still a scrawny beginner, the data from the study at hand does suggest that there will be an increase in the marginal utility of your workouts after one or two weeks of detraining. Moreover, there is no reason to be afraid that you could lose muscle within this short time period. Based on the absolute numbers in the study at hand (cf. figure 2), you could rather expect to see a non-significant increase in muscle mass that will occur during shorter (1-2 weeks) detraining phases.

      If you have no idea what macro- and micro-cycles are or are clueless about how to incorporate phases of detraining and - as a possible alternative with potentially similar effects - tapering into your routine, I suggest you go back to part VI of the Step by Step Guide to Your Own Workout
      Just don't forget, that just as it was the case in the previously mentioned 2012 human study by Ogasawara et al., you will see and feel the beneficial effects of the detraining period, not before you are back on the grind for 1-2 weeks. If you look back at the protocol, Ogasawara et al. used, the fact that the gains were "just" identical (even that would be a huge plus: after all you get the added bonus of reduced risk of injury, overtraining, etc. without missing out on a single additional mm on your arms, chest, shoulders, quads, hams, and what not) would actually support my gut feeling that a detraining period (= no training at all) of three weeks after "only" 6 weeks, could be a little too long. In the study at hand, which is obviously not a human stud and did not involve a regular full-body split routine, it did after all take no more than 12 days, i.e. 9 days less for the signaling responses to return to baseline.

      Further speculations about optimal off-times and respective increases in muscle gains would be mere speculation, so that I would suggest, we will postpone more concrete suggestions until the next paper from Ogosawara's group at the Research Organization of Science and Technology to be published. If we assume that it will take another 3 months, which happens to be the interval between the aforementioned human study and the study at hand, you better mark the first two weeks of April 2013 in your calendar, if you don't want to miss the respective SuppVersity post on the matter ;-)



      Regular periods of detraining are not the only thing you should keep in mind, when you lay out your workout routine. A study I wrote about in June 2012, for example showed that appropriate periodization can help you "Cut 12% Body Fat in 12 Weeks, Get Stronger, Bigger and Better Conditioned" (read more)
      In short, what is it, this paper brings to the table and what are the implications? The "new" information this paper has to offer pertains to the restorative effects of detraining n the exercise induced protein signaling cascade that will eventually result in skeletal muscle hypertrophy (=muscle gains). The differential protein expression in the different training groups tell us, that the 12 days of detraining effectively restored the p70S6K an  rpS96 that declines with each and every workout during periods of continuous training.

      The practical implications of these findings, should actually be obvious: Incorporating regular periods of detraining in your macrocycles and most importantly sticking to the plan, will not just help you to avoid overtraining with all it's negative side effects, it will also prevent the hypertrophy response to your workouts from diminishing and thus propel your lean mass gains.
      Update: Steven Acerra called my attention to an older study by Kadi et al. who report distinct effects of detraining on the satellite cell response to heavy resistance training (Kadi. 2004). According to the results of their study, the often overlooked contribution of the satellite cells to the structural underpinnings of skelatal muscle appears to peak early in the detraining phase with the maximal number of satellite cells per muscle fiber being achieved after 10 days of detraining and a return to pre-training levels after 90 days. Despite the fact that a straight forward extrapolation of practical recommendations based on these observations alone is not feasible, the researchers observations do confirm that there is a multilayered benefit to short (~14 day) detraining periods.
      Whether 12 days is the "optimal" length for a detraining period, if this "optimum" depends on the length, intensity and type of the previous training period, whether it's body part specific (like stop training arms for two weeks to restore your growth response in the biceps and triceps) and whether or not the training status of an individual has any impact on the time that's necessary for the "reset" to take place, will have to be elucidated in previous studies. That the SuppVersity is going to be the place, where you will read about these first, is something I probably don't have to tell you, right?

      References: 
      • Coffey VG, Zhong Z, Shield A, Canny BJ, Chibalin AV, Zierath JR, and Hawley JA. Early signaling responses to divergent exercisestimuli in skeletal muscle from well-trained humans. FASEB J. 2006; 20: 190-192. 
      • Kadi F, Schjerling P, Andersen LL, Charifi N, Madsen JL, Christensen LR, Andersen JL. The effects of heavy resistance training and detraining on satellite cells in human skeletal muscles. J Physiol. 2004 Aug 1;558(Pt 3):1005-12. Epub 2004 Jun 24.
      • Ogasawara R, Yasuda T, Sakamaki M, Ozaki H, Abe T. Effects of periodic and continued resistance training on muscle CSA and strength in previously untrained men. Clin Physiol Funct Imaging. 2011 Sep;31(5):399-404.
      • Ogasawara R, Kobayashi K, Tsutaki A, Lee K, Abe T, Fujita S, Nakazato K, Ishii N. mTOR signaling response to resistance exercise is altered by chronic resistance training and detraining in skeletal muscle. J Appl Physiol. 2013 Jan 31. [Epub ahead of print]