Skeletal muscle is a highly organized tissue that requires cooperation of many different structures and components for proper function. We explored the use of a flexor digitorum brevis (FDB) myofiber culture system to better model highly differentiated aspects of skeletal muscle in an in vitro system. Indirect immunofluorescence of FDB myofibers allowed us to better determine the subcellular localization of KLHL41, a new nemaline myopathy (NM) gene product, to ER-like subdomains of the sarcoplasmic retiuculum. By comparing FDB myofibers from wild type and myotubularin knockout mice with X-linked myotubular myopathy (XLMTM), we were also able to analyze satellite cell populations, showing that the knockout mice suffered a marked decrease in associated myogenic satellite cells. This supports concurrent data from our lab indicating a disease progression-related increase in apoptosis and a decrease in satellite cell proliferation in XLMTM.

Ce travail de thèse vise à améliorer notre compréhension des effets l'entraînement en résistance sur la performance et le muscle strié squelettique. La dynamique de ces effets de l'entraînement a été appréhendée de façon systématique grâce à des outils issus de la théorie des systèmes, auprès de 26 rongeurs entraînés en résistance dans un protocole d'escalade avec charges additionnelles. Le modèle classique (Banister et coll, 1975) a permis de décrire les variations de performance de manière significative (R2 = 0,53, P<0,001). L'origine des gains de performance très marqués (+136% par rapport au groupe contrôle) a été recherchée parmi les mécanismes adaptatifs musculaires potentiels. A l'issue de l'entraînement, une augmentation de l'activité de la myosine ATPase de 123 ± 61% indépendante du phénotype a été observée par rapport aux animaux contrôles. Cette augmentation de la puissance chimique consommée semble liée à une augmentation de la vitesse des étapes d'hydrolyse de l'ATP et surtout de celle de la libération des produits de cette hydrolyse (i.e. ADP et Pi) accompagnant la bascule de la tête de myosine. Une nouvelle forme de plasticité musculaire semble avoir été identifiée. Sur la base des mécanismes adaptatifs musculaires, une nouvelle formulation mathématique plus physiologique du modèle des effets de l'entraînement a été proposée et a aboutit à une meilleure qualité d'ajustement (R2 = 0,71, P<0,001). La fonction impulsionnelle du modèle classique a été remplacée par une fonction exponentielle de croissance qui semble plus appropriée pour rendre compte à la fois des variations de performance mais aussi des adaptations qui surviennent au sein du tissu musculaire comme au sein des unités contractiles elles-mêmes. / This thesis work aims to improve our understanding of the effects of resistance training on performance and skeletal muscle. The dynamic of these effects of training has been apprehended systematically trough tools from systems theory, with 26 rodents resistance trained on a climbing protocol with additional weights. The classical model (Banister et al, 1975) was suitable to analyze the training response (R2 = 0.53, P <0.001). The origin of the very marked performance gains (+ 136% compared to the control group) was investigated among the potential muscle adaptive mechanisms. At the end of the training program, an increase of 123 ± 61% in myosin ATPase activity independent of the phenotype was observed compared to control animals. This increase in myosin ATPase activity seems to occur precisely during the main myosin head isomerization step (i.e. powerstroke) that includes the liberation of the hydrolysis products, and to a lesser extent, during ATP hydrolysis step. A new form of muscular plasticity seems identified. Based on muscle adaptive mechanisms, a new mathematical formulation, more physiological, of the model of the training effects has been proposed and resulted in a better fit (R2 = 0.71, P <0.001). The impulse function of the traditional model has been replaced by an exponential growth function that seems more suitable to analyze both the training response and the adaptations that occur within the muscle tissue as in the contractile units themselves.

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<p dir="ltr">Skeletal muscle accounts for 40% of total body weight and the homeostasis of muscle tissue is critical in maintaining proper body function. Skeletal muscle develops during the embryonic stages from the muscle progenitor cells derived from the dermomyotome structure. The myogenic progenitor cells contribute to the primary myogenesis by forming the primary myotubes which are the founding structures that the secondary myogenesis continues to build on. A portion of the myogenic progenitor cells makes up the adult muscle stem cells residing in homeostatic muscle tissue. The adult muscle stem cells contribute substantially for the adult muscle regeneration. Due to the significance of the muscle tissue and the importance of muscle stem cells, dysregulation of the muscle homeostasis or the muscle stem cell homeostasis will result in severe pathological conditions such as myopathy.</p><p dir="ltr">Mitochondria are cellular organelles that are responsible for generating energy needed for cellular processes, especially for muscle tissue where muscle contraction requires the presence of ATP. On the other hand, mitochondria also serve as signaling molecules and provide macromolecules for the biosynthesis. FAM210A (Family With Sequence Similarity 210 Member A) protein was shown to impact the lean mass of human subjects yet a detailed study on the effect of FAM210A in skeletal muscle was not performed, nor has the molecular mechanisms through which FAM210A function been elucidated. Therefore, I take on the task to unveil the function of FAM210A in muscle development, muscle homeostasis and muscle stem cell behavior by using a combination of mouse models with different myogenic promoters to target <i>Fam210a</i> at different developmental stages.</p><p dir="ltr">In the first part of the thesis, I investigated the role of FAM210A in post differentiation myofibers. Using the <i>Myl1</i>^<em>Cre</em> driven deletion of <i>Fam210a</i>, I found that <i>Fam210a</i>^<em>MKO</em> had normal development before 3 weeks of age, but the growth was stagnant from 4 weeks on, and the mice did not survive past 8 weeks of age. I found that the assembly of the ribosomes in the <i>Fam210a</i>^<em>MKO</em> was defective, leading to impaired translation which attenuated the muscle atrophy phenotype. I identified through proteomics that the mitochondrial autophagy and proteostatic control pathways were significantly induced yet mitochondrial organization and energetic proteins were downregulated. Metabolomics analysis showed that the signaling metabolite acetyl-CoA was increased in the <i>Fam210a</i>^<em>MKO</em> which led to increased protein acetylation, specifically, we showed that the ribosomal proteins were hyperacetylated, and that the acetylation increase was elicited by the <i>Fam210a</i>-null mitochondria.</p><p dir="ltr">In the second part of the thesis, I investigated the function of FAM210A in muscle progenitor cells. In the <i>FamMKO</i> mice, I found that deletion of <i>Fam210a</i> from embryonic myogenic progenitor cells led to developmental arrest and postnatal death at day 6. In the <i>FamPKO</i> mice, I found that <i>Fam210a</i> is needed for adult muscle stem cell to contribute to regeneration. Loss of <i>Fam210a</i> leads to the regenerative defects when the muscle was exposed to injury cues. We further showed that <i>Fam210a</i> deletion in muscle stem cells resulted in disruption of the proteostatic control over muscle stem cell activation, thereby forbidding the translational increase necessary to facilitate activation and proliferation. Furthermore, I showed that <i>Fam210a</i> deletion leads to excessive OPA1 cleavage, which contributes to the regenerative failure of muscle stem cells as fusion is required for the mitochondrial network remodeling during regeneration. Therefore, <i>Fam210a</i> safeguards the mitochondrial network and proteostasis during regeneration.</p><p dir="ltr">In summary, my studies characterized the functional contribution of FAM210A during embryonic muscle development, muscle mass maintenance and adult muscle stem cell homeostasis. The regulation of FAM210A in these three processes impinge on the translational regulation. My studies further demonstrated the importance of mitochondrial regulated protein translation in skeletal muscle and muscle stem cells.</p>