Original article
The Effect of 30 Minutes of Passive Stretch of the Rat Soleus Muscle on the Myogenic Differentiation, Myostatin, and Atrogin-1 Gene Expressions

Presented as abstracts to the Federação de Sociedades de Biologia Experimental, August 25–28, 2004, Águas de Lindóia, Brazil, and the American Physiological Society, October 6–9, 2004, Austin, TX.
https://doi.org/10.1016/j.apmr.2005.08.126Get rights and content

Abstract

Gomes AR, Soares AG, Peviani S, Nascimento RB, Moriscot AS, Salvini TF. The effect of 30 minutes of passive stretch of the rat soleus muscle on the myogenic differentiation, myostatin, and atrogin-1 gene expressions.

Objective

To evaluate the effect of passive stretch, applied for 30 minutes to the rat soleus muscle, on the myogenic differentiation (myoD), myostatin, and atrogin-1 gene expressions.

Design

Case-controlled study.

Setting

University laboratory.

Animals

Fifty 12-week-old male Wistar rats.

Interventions

Six groups of animals were given a single stretch bout and were evaluated immediately and 8, 24, 48, 72, and 168 hours later. Another 3 groups were evaluated immediately after 2, 3, and 7 stretches. An intact control group was also analyzed.

Main Outcome Measures

The messenger ribonucleic acid (mRNA) levels of myoD, myostatin, and atrogin-1 were assessed by real-time polymerase chain reaction.

Results

Twenty-four hours after a single session of stretch only, the myoD mRNA levels had increased compared with the control group, whereas an increase in the atrogin-1 expression was observed after 2, 3, and 7 stretches.

Conclusions

A single session of passive stretch increased the myoD gene expression, a factor related to muscle growth. Interestingly, daily stretches increased the atrogin-1 gene expression, a gene primarily associated with muscle atrophy. The results indicated that gene expression was responsive to the number of stretch sessions.

Section snippets

Animal Care and Experimental Groups

We used 50 male, 3-month-old Wistar rats (weight, 373±32g). They were housed in plastic cages in a room with controlled environmental conditions and had free access to water and standard food. This study was conducted in accordance with the university approval for the care and use of laboratory animals. The rats were anesthetized by an intraperitoneal injection of xylazine (12mg/kg) and ketamine (95mg/kg) for the stretching of the soleus muscle and muscle dissection. Afterward, they were killed

Muscle Mass

No difference was found in the weight of the soleus muscles among any groups evaluated (table 1).

MyoD Gene Expression

An increase in the myoD gene expression was found 24 hours after a single session of stretch, when compared with the control group (3.4±0.9-fold vs 1±0.06-fold, respectively, ANOVA, P=.001; fig 1A). Subsequently, the myoD gene expression returned to the control levels for all the times evaluated (48h, 72h, 168h). On the other hand, when daily sessions of stretch were performed, the myoD levels

Discussion

The results of this study show that a single session of passive stretch, performed for 30 minutes, induced a 300% increase in myoD mRNA when compared with the control soleus muscle, but did not change the gene expressions of myostatin and atrogin-1. As described in the literature, muscles immobilized in a lengthened position by a plaster cast, present an increase in the myoD gene expression.11, 16, 17, 45 However, in these earlier studies, the muscles were maintained immobilized in a stretched

Conclusions

A single session of passive stretch for 30 minutes increased myoD gene expression, suggesting a hypertrophic effect. However, daily sessions of stretch blocked this effect and increased atrogin-1 gene expression, which has been primarily associated with muscle atrophy. Thus, passive stretch alters both hypertrophic and atrophic skeletal muscle mechanisms. Also, the number of stretch sessions was determinant in both atrogin-1 and myoD responsiveness.

Acknowledgment

We thank Marcia Gislene, BSc, for providing technical assistance.

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    Supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (grant nos. 01/13523-4, 01/135221-1, 03/10889-3), Coordenção de Aperfeiçoamento de Pessoal de Nivel Superior (grant no. 33001014016P7), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (grant no. 501737/2004-9).

    No commercial party having a direct financial interest in the results of the research supporting this article has or will confer a benefit upon the authors or upon any organization with which the authors are associated.

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