“MIGHTY MOUSE” Gene Increases Muscle Mass in Humans

By studying the genes of a German child born with unusually well developed muscles, an international research team has discovered the first evidence that the gene whose loss makes “mighty mice” also controls muscle growth in people.

Writing in the June 24 issue of the New England Journal of Medicine, German neurologist Markus Schuelke, M.D., and the team show that the child’s extra-large muscles are due to an inherited mutation that effectively silences the myostatin gene, proving that its protein normally keeps muscle development in check in people.

People with muscle-wasting conditions such as muscular dystrophy, and others just wanting to “bulk up,” have eagerly followed work on myostatin, hoping for a way to counteract the protein’s effects in order to build or rebuild muscle mass. But while research with mice has continued to reveal myostatin’s role and the effects of interfering with it, no one knew whether any of the results would be relevant to humans.

“This is the first evidence that myostatin regulates muscle mass in people as it does in other animals,” says Se-Jin Lee, M.D., Ph.D., professor of molecular biology and genetics in the Institute for Basic Biomedical Sciences at Johns Hopkins and co-author on the study. “That gives us a great deal of hope that agents already known to block myostatin activity in mice may be able to increase muscle mass in humans, too.”

Lee and his team discovered in 1997 that knocking out the myostatin gene led to mice that were twice as muscular as their normal siblings, lending them the moniker “mighty mice.” Later, others showed that naturally bulky cattle, such as Belgian Blues, got their extra muscles from lack of myostatin, too.

An unusual opportunity to examine myostatin’s role in humans arose when Schuelke examined a newborn baby boy, almost five years ago, and was struck by the visible muscles on the infant’s upper legs and upper arms. When ultrasound proved that the muscles were roughly twice as large as other infants’, but otherwise normal, Schuelke realized that a naturally occurring mutation in the child’s myostatin gene might be the cause.

Sequencing the myostatin gene from the boy and his mother, who had been a professional athlete, revealed a single change in the building blocks of the gene’s DNA. Surprisingly, the change was not in the gene regions that correspond to the resulting protein, but in the intervening regions that are used only to create protein-making instructions, thus changing the gene’s protein-building message.

“The mutation caused the gene’s message, the messenger RNA, to be wrong,” says Hopkins

neurologist Kathryn Wagner, M.D., Ph.D., who tested the genetic mutation’s effect in laboratory studies. “If the message had been used to make a protein, it would be much shorter than it should be. But we think the process doesn’t even get that far; instead the cells just destroy the message.”

Co-authors from Wyeth Research, Cambridge, Mass., analyzed samples of the child’s blood for evidence of the myostatin protein and found none. “Both copies of the child’s myostatin gene have this mutation, so little if any of the myostatin protein is made,” says Schuelke. “As a result, he has about twice the muscle mass of other children.”

Completely lacking myostatin, the boy is stronger than other children his age, and fortunately has no signs of problems with his heart so far, Schuelke says. But he adds that it’s impossible to know whether the lack of myostatin in that crucial muscle might lead to problems as the boy gets older.

While other family members — the boy’s mother and her brother, father and grandfather — were also reported to have been usually strong, only the mother’s DNA was available for analysis along with her son’s. Schuelke discovered that only one copy of the mother’s myostatin gene had the mutation found in both copies of her son’s myostatin gene. (We have two copies of each gene; one inherited from the mother and one inherited from the father.)

—————————-
Article adapted by MD Sports Weblog from original press release.
—————————-

 Contact: Joanna Downer
Johns Hopkins Medical Institutions

 

The Johns Hopkins researchers were funded by the National Institutes of Health and the Muscular Dystrophy Association. The German researchers were funded by the parents’ self-help group (Helft dem muskelkranken Kind).

Authors on the paper are Schuekle, Christoph Hubner, Thomas Riebel and Wolfgang Komen of Charite, University Medical Center Berlin, Germany; Wagner and Lee of Johns Hopkins; Leslie Stolz and James Tobin of Wyeth Research, Cambridge, Ma.; and Thomas Braun of Martin-Luther-University, Halle-Wittenberg, Germany.

*Under a licensing agreement between MetaMorphix Inc. and The Johns Hopkins University, Lee is entitled to a share of royalty received by the University on sales of products described in this article. Lee also is entitled to a share of sublicensing income from arrangements between MetaMorphix and American Home Products (Wyeth Ayerst Laboratories) and Cape Aquaculture Technologies. Lee and the University own MetaMorphix Inc. stock, which is subject to certain restrictions under University policy. Lee owns Cape Aquaculture Technologies stock, which is subject to certain restrictions under University policy. Lee has served as a paid consultant to MetaMorphix Inc. The terms of these arrangements are being managed by The Johns Hopkins University in accordance with its conflict of interest policies.

Super protein creates fat burning Metabolic ’shield” against weight gain…

And it increases endurance to run a mile and decreases inflammation

The Salk Institute scientist who earlier discovered that enhancing the function of a single protein produced a mouse with an innate resistance to weight gain and the ability to run a mile without stopping has found new evidence that this protein and a related protein play central roles in the body’s complex journey to obesity and offer a new and specific metabolic approach to the treatment of obesity related disease such as Syndrome X (insulin resistance, hyperlipidemia and atherosclerosis).

Dr. Ronald M. Evans, a Howard Hughes Medical Investigator at The Salk Institute’s Gene Expression Laboratory, presented two new studies (date) at Experimental Biology 2005 in the scientific sessions of the American Society for Biochemistry and Molecular Biology. The studies focus on genes for two of the nuclear hormone receptors that control broad aspects of body physiology, including serving as molecular sensors for numerous fat soluble hormones, Vitamins A and D, and dietary lipids.

The first study focuses on the gene for PPARd, a master regulator that controls the ability of cells to burn fat. When the “delta switch” is turned on in adipose tissue, local metabolism is activated resulting in increased calorie burning. Increasing PPARd activity in muscle produces the “marathon mouse,” characterized by super-ability for long distance running. Marathon mice contain altered muscle composition, which doubles its physical endurance, enabling it to run an hour longer than a normal mouse. Marathon mice contain increased levels of slow twitch (type I) muscle fiber, which confers innate resistance to weight gain, even in the absence of exercise.

Additional work to be reported at Experimental Biology looks at another characteristic of PPARd: its role as a major regulator of inflammation. Coronary artery lesions or atherosclerosis are thought to be sites of inflammation. Dr. Evans found that activation of PPARd suppresses the inflammatory response in the artery, dramatically slowing down lesion progression. Combining the results of this new study with the original “marathon mouse” findings suggests that PPARd drugs could be effective in controlling atherosclerosis by limiting inflammation and at the same time promoting improved physical performance.

Dr. Evans says he is very excited about the therapeutic possibilities related to activation of the PPARd gene. He believes athletes, especially marathon runners, naturally change their muscle fibers in the same way as seen in the genetically engineered mice, increasing levels of fat-burning muscle fibers and thus building a type of metabolic ’shield” that keeps them from gaining weight even when they are not exercising.

But athletes do it through long periods of intensive training, an approach unavailable to patients whose weight or medical problems prevent them from exercise. Dr. Evans believes activating the PPARd pathway with drugs (one such experimental drug already is in development to treat people with lipid metabolism) or genetic engineering would help enhance muscle strength, combat obesity, and protect against diabetes in these patients.

—————————-
Article adapted by MD Sports Weblog from original press release.
—————————-

Contact: Sarah Goodwin
Federation of American Societies for Experimental Biology

Creatine/CLA combo enhances muscle size and decreases body fat

Lower muscle mass and an increase in body fat are common consequences of growing older.

While exercise is a proven way to prevent the loss of muscle mass, a new study led by McMaster researcher Dr. Mark Tarnopolsky shows that taking a combination of creatine monohydrate (CrM) and conjugated linoleic acid (CLA) in addition to resistance exercise training provides even greater benefits.

The study to be published on Oct. 3 in PLoS One, an international, peer-reviewed online journal of the Public Library of Science, involved 19 men and 20 women who were 65 years or older and took part in a six-month program of regular resistance exercise training.

In the randomized double blind trial, some of the participants were given a daily supplement of creatine (a naturally produced compound that supplies energy to muscles) and linoleic acid (a naturally occurring fatty acid), while others were given a placebo. All participants took part in the same exercise program.

The exercise training resulted in improvements of functional ability and strength in all participants, but those taking the CrM and CLA showed even greater gains in muscle endurance, an increase in fat-free mass and a decrease in the percentage of body fat.

“This data confirms that supervised resistance exercise training is safe and effective for increasing strength and function in older adults and that a combination of CrM and CLA can enhance some of the beneficial effects of training over a six month period,” said Tarnopolsky, a professor of pediatrics and medicine.

This study provides functional outcomes that build on an earlier mechanistic study co-led by Tarnopolsky and Dr. S. Melov at the Buck Institute of Age Research, published in PLoS One this year, which provided evidence that six months of resistance exercise reversed some of the muscle gene expression abnormalities associated with the aging process.

—————————-
Article adapted by MD Sports Weblog from original press release.
—————————-

Contact: Veronica McGuire
McMaster University

Quadrupling of Muscle Mass Without Steroids

Myostatin (MSTN) is a transforming growth factor-ß (TGF-ß) family member that plays a critical role in regulating skeletal muscle mass [1]. Mice engineered to carry a deletion of the Mstn gene have about a doubling of skeletal muscle mass throughout the body as a result of a combination of muscle fiber hyperplasia and hypertrophy [2]. Moreover, loss of myostatin activity resulting either from postnatal inactivation of the Mstn gene [3], [4] or following administration of various myostatin inhibitors to wild type adult mice [5]–[7] can also lead to significant muscle growth. Hence, myostatin appears to play as least two distinct roles, one to regulate the number of muscle fibers that are formed during development and a second to regulate growth of muscle fibers postnatally. The function of myostatin appears to have been conserved across species, as inactivating mutations in the myostatin gene have been demonstrated to cause increased muscling in cattle [8]–[11] , sheep [12], dogs [13] and humans [14]. As a result, there has been considerable effort directed at developing strategies to modulate myostatin activity in clinical settings where enhancing muscle growth may be beneficial. In this regard, loss of myostatin activity has been demonstrated to improve muscle mass and function in dystrophic mice [15]–[17] and to have beneficial effects on fat and glucose metabolism in mouse models of obesity and type II diabetes [18].

Myostatin is synthesized as a precursor protein that undergoes proteolytic processing to generate an N-terminal propeptide and a C-terminal dimer, which is the biologically active species. Following proteolytic processing, the propeptide remains bound to the C-terminal dimer and maintains it in an inactive, latent complex [6], [19], [20], which represents one of the major forms of myostatin that circulates in the blood [21], [22]. In addition to the propeptide, other binding proteins are capable of regulating myostatin activity in vitro, including follistatin [19], [21], FLRG [22], and Gasp-1 [23]. We previously showed that follistatin can also block myostatin activity in vivo; specifically, we showed that follistatin can ameliorate the cachexia induced by high level expression of myostatin in nude mice [21] and that transgenic mice expressing follistatin in muscle have dramatic increases in muscle mass [19]. Here, I show that overexpression of follistatin can also cause substantial muscle growth in mice lacking myostatin, demonstrating that other TGF-ß related ligands normally cooperate with myostatin to suppress muscle growth and that the capacity for enhancing muscle growth by targeting this signaling pathway is much larger than previously appreciated.

Results

Increased muscle mass in transgenic mice expressing FLRG

Previous studies have identified several proteins that are normally found in a complex with myostatin in the blood [22], [23]. One of these is the follistatin related protein, FLRG, which has been demonstrated to be capable of inhibiting myostatin activity in vitro. To determine whether FLRG can also inhibit myostatin activity in vivo, I generated a construct in which the FLRG coding sequence was placed downstream of a myosin light chain promoter/enhancer. From pronuclear injections of this construct, a total of four transgenic mouse lines (Z111A, Z111B, Z116A, and Z116B) were obtained containing independently segregating insertion sites. Each of these four transgenic lines was backcrossed at least 6 times to C57 BL/6 mice prior to analysis in order to control for genetic background effects. Northern analysis revealed that in three of these lines the transgene was expressed in skeletal muscles but not in any of the non-skeletal muscle tissues examined (Figure 1); in the fourth line, Z111B, the expression of the transgene was below the level of detection in these blots. As shown in Table 1, all four lines exhibited significant increases in muscle weights compared to wild type control mice. These increases were observed in all four muscles that were examined as well as in both sexes. Moreover, the rank order of magnitude of these increases correlated with the rank order of expression levels of the transgene; in the highest-expressing line, Z116A, muscle weights were increased by 57–81% in females and 87–116% in males compared to wild type mice. Hence, FLRG is capable of increasing muscle growth in a dose-dependent manner when expressed as a transgene in skeletal muscle.

The research was funded by grants from the NIH and the Muscular Dystrophy Association and by a gift from Merck Research Laboratories.

See http://www.jhu.edu/sejinlee/%20for%20more%20information for more information.
Citation: Lee S-J (2007) Quadrupling Muscle Mass in Mice by Targeting TGF-ß Signaling Pathways. PLoS ONE 2(8): e789. doi:10.1371/journal.pone.0000789

LINK TO THE PUBLISHED ARTICLE http://www.plosone.org/doi/pone.0000789

Source: Nick Zagorski
Johns Hopkins Medical Institutions

Caffeine intake negates the benefits of creatine supplements

The serious athlete knows better than to rely just on a famous cereal to provide additional energy in preparation of a sporting event. Supplements have assumed an important role in today’s training regimen. Some – such as anabolic steroids — have been deemed illegal by most sports authorities. Others – such as caffeine and creatine — are controversial yet presently allowed.Background
Caffeine, the primary ingredient of coffee, is used as a central nervous system stimulant, diuretic, circulatory and respiratory stimulant, and as an adjunct in the treatment of headaches. Evidence shows that caffeine intensifies muscle contractions, masks the discomfort of physical exertion, and even speeds up the use of the muscles’ short-term fuel stores. Some exercise physiologists believe that caffeine might improve performance by increasing fat oxidation and conserving muscle glycogen.

Creatine is used by athletes to increase lean body mass and improve performance in single and repetitive high-intensity, short-duration exercise tasks such as weightlifting, sprinting, and cycling. It is a popular nutritional supplement that is used by physically active people – from recreational exercisers to Olympic and professional athletes. According to a recent survey, 28 percent of athletes in an NCAA Division IA program reported using creatine. The creatine that is normally present in human muscle may come from two potential sources: dietary (animal flesh) and internally manufactured.

The purpose of creatine supplementation is to increase either total creatine stores or phosphocreatine (PCr) stores within muscle. Supplementation increases the rate of resynthesis of creatine phosphate following exercise. Various studies have shown increased muscle PCr levels after supplementing with 20-30 grams of creatine monohydrate daily.

Creatine supplementation has also been known to shorten relaxation time during intermittent maximal iosometric muscle contraction. This shortened time, coupled with a creatine loaded muscle facilitates calcium absorption into the sarcoplasmic reticulum (the endoplasmic reticulum of skeletal and cardiac muscle). However, some believe that caffeine intake enhances calcium release from the sarcoplasmic reticulum.

The Study
This has lead a research team from Belgium to suggest that the combined effects of creatine and caffeine supplementation may be counterproductive to creatine’s effect on muscle relaxation time. The authors of the study, “Opposite Actions of Caffeine and Creatine on Muscle Relaxation Time in Humans” are P. Hespel, B. Op ‘T Eijnde, and M. Van Leemputte, all from the Department of Kinesiology, Katholieke Universiteit Leuven, Leuven, Belgium. Their findings appear in the February 2002 edition of the Journal of Applied Physiology.

Methodology
Ten physical education students (nine men and one woman) participated in the study. They were told to abstain from medication and caffeine intake one week prior to the experiment. The subjects were additionally asked to avoid changes in their level of physical activity and diet during the 25-week duration of the study. In this double blind experiment, the subjects performed the exercise test before and after creatine supplementation, short-term caffeine intake, creatine supplementation in the short term, acute caffeine intake, or a placebo.

This study required the random assignment of the students into five experimental protocols, each lasting eight days. Three elements were measured during an experiment consisting of 30 intermittent contractions of quadriceps entailing two seconds of stimulation and two seconds of rest. Measurements included maximum torque (Tmax), contraction time (CT) from 0.25 to 0.75 of Tmax, and relaxation time (RT) from 0.75 to 0.25 of max.

Results
Key findings of this study included:

· a confirmation of the fact that oral creatine supplementation shortens muscle relaxation time in humans: relation time was reduced by five percent and was significantly shorter than after the placebo;

· discovery that the intake of caffeine, combined with a daily creatine supplement, counteracted the beneficial effects of creatine intake on relaxation time and fatigue enhanced this inhibitory effect; and

· the observation that caffeine reduces the functional capacity of sacroplasmic reticulum calcium ATPase.

Conclusion The researchers believe that the findings from this experiment offer indirect evidence that suggests that facilitation of muscle relaxation may be important to the ergogenic action of creatine supplementation as well as power production during sprint exercises.

However, for the athlete in training, the key finding is that sustained caffeine intake, over a three-day period, negates the benefits of creatine supplements.

—————————-
Article adapted by MD Only Sports Weblog from original press release.
—————————-

Contact: Donna Krupa
American Physiological Society

Majority of Steroid Users Are Muscle Seeking Adults, Not Athletes

The majority of non-medical anabolic-androgenic steroid (AAS) users are not cheating athletes or risk-taking teenagers. According to a recent survey, containing the largest sample to date and published in the online open access publication, Journal of the International Society of Sports Nutrition, the typical male user is about 30 years old, well-educated, and earning an above-average income in a white-collar occupation. The majority did not use steroids during adolescence and were not motivated by athletic competition or sports performance.

The study, conducted by a collaboration of researchers from around the country coordinated by Jason Cohen, Psy.D. candidate, used a web-based survey of nearly 2,000 US males. Whereas athletes are tempted to take anabolic steroids to improve sports performance, the study suggests that physical self-improvement motivates the unrecognized majority of non-medical AAS users who particularly want to increase muscle mass, strength, and physical attractiveness. Other significant but less highly ranked factors included increased confidence, decreased fat, improved mood and attraction of sexual partners.

Although often considered similar to abusers of narcotics and other illicit drugs (e.g., heroin or cocaine), non-medical AAS users are remarkably different. These users follow carefully planned drug regimens in conjunction with a healthy diet, ancillary drugs and exercise. As opposed to the spontaneous and haphazard approach seen in abusers of psychotropic drugs, everything is strategically planned to maximize benefits and minimize harm. “This is simply not a style or pattern of use we typically see when we examine substance abuse” said Jack Darkes, Ph.D., one of the authors. “The notions of spontaneous drug seeking and loss of control do not apply to the vast majority of AAS users,” added co-author Daniel Gwartney, M.D.

“These findings question commonly held views of typical AAS users and their underlying motivations,” said Rick Collins, one of the study’s authors. “The focus on ‘cheating’ athletes and at risk youth has led to irrelevant policy as it relates to the predominant group of non-medical AAS users. The vast majority of AAS users are not athletes and hence, are not likely to view themselves as cheaters. The targeting of athletes through drug testing and other adolescent or sports-based interventions has no bearing on non-competitive adult users.” The study concludes that these AAS users are a driven and ambitious group dedicated to gym attendance, diet, occupational goals and educational attainment. “The users we surveyed consider that they are using directed drug technology as one part of a strategy for physical self-improvement within a health-centered lifestyle,” said Collins. “Effective public policy should begin by accurately identifying who’s using steroids and why. We hope our research – the largest adult survey of non-medical AAS use we know of – is a significant step forward in that direction.”

—————————-
Article adapted by MD Only Sports Weblog from original press release.
—————————-

Contact: Charlotte Webber
BioMed Central

Article:
A League of Their Own: Demographics, Motivations and Patterns of Use of 1,955 Male Adult Non-Medical Anabolic Steroid Users in the United States
Jason Cohen, Rick Collins, Jack Darkes and Dan Gwartney
Journal of the International Society of Sports Nutrition (in press)

During embargo, article available at: http://www.jissn.com/imedia/1374735248154681_article.pdf?random=454689

After the embargo, article available from the journal website at: http://www.jissn.com