Claude Louis Gallien

Regulation by thyroid hormones of terminal differentiation in the skeletal dorsal muscle. I: Neonate mouse

Changes both in the ATPase myofibrillar profile and in the electrophoretic pattern of myosin isoforms were examined in the mouse dorsal skeletal muscle (longissimus) during postnatal development. In the newborn, only type II C and a few type I fibers were present; differentiation into type II A and II B fibers took place during the 3 weeks following birth. During the same period, a transition from three neonatal isomyosins to four adult isoforms was observed. The two phenomena were related to a marked increase in the serum thyroid hormones levels. Hypothyroidism and hyperthyroidism experiments were performed. Hypothyroidism produced by propylthiouracil treatment of pregnant females and thiourea injections of the litters was shown to induce a complete inhibition of postnatal muscular differentiation. Hyperthyroidism produced by triiodothyronine treatment of the neonate mice significantly accelerated the myosin transition and the switch in the myofibrillar pattern. Our results suggest a primordial role for thyroid hormones in directly regulating the appearance of myosin and fiber adult types and in modulating directly or indirectly the disappearance of the neonatal types.

Expression and neural control of follistatin versus myostatin genes during regeneration of mouse soleus

Follistatin and myostatin are two secreted proteins involved in the control of muscle mass during development. These two proteins have opposite effects on muscle growth, as documented by genetic models. The aims of this work were to analyze in mouse, by using in situ hybridization, the spatial and temporal expression patterns of follistatin and myostatin mRNAs during soleus regeneration after cardiotoxin injury, and to investigate the influence of innervation on the accumulation of these two transcripts. Follistatin transcripts could be detected in activated satellite cells as early as the first stages of regeneration and were transiently expressed in forming myotubes. In contrast, myostatin mRNAs accumulated persistently throughout the regeneration process as well as in adult control soleus. Denervation significantly affected both follistatin and myostatin transcript accumulation, but in opposite ways. Muscle denervation persistently reduced the levels of myostatin transcripts as early as the young myotube stage, whereas the levels of follistatin mRNA were strongly increased in the small myotubes in the late stages of regeneration. These results are discussed with regard to the potential functions of both follistatin, as a positive regulator of muscle differentiation, and myostatin, as a negative regulator of skeletal muscle growth. We suggest that the belated up‐regulation of the follistatin mRNA level in the small myotubes of the regenerating soleus as well as the down‐regulation of the myostatin transcript level after denervation contribute to the differentiation process in denervated regenerating muscle. Developmental Dynamics 227:256–265, 2003.

Regulation by thyroid hormones of terminal differentiation in the skeletal dorsal muscle: II. Urodelan amphibians☆

In the urodelan amphibian Pleurodeles waltlii, spontaneous anatomical metamorphosis was correlated with an increase in the serum level of thyroxine (T4). It was also accompanied by a change in the myofibrillar ATPase profile of the dorsal skeletal muscle; fibers of larval type were gradually replaced by the adult fiber types I, II A, and II B. Likewise, a myosin isoenzymic transition was observed in dorsal muscle, larval isomyosins were replaced by adult isoforms. In a related species, Ambystoma mexicanum, in which no spontaneous external metamorphosis occurs under standard conditions, the serum T4 level was shown to remain low. During further development, the myofibrillar ATPase profile acquired the adult fiber types, but a high percentage of immature fibers of type II C persisted. Myosin isoenzymic transition was also incomplete; larval isoforms were still distinguished in the neotenic adults. In experimental hypothyroidian P. waltlii, no external metamorphosis occurred; the myofibrillar ATPase profile was of the immature type, and the larval isomyosins persisted. Triiodothyronine induced experimental anatomical metamorphosis in A. mexicanum; only limited changes in the myofibrillar ATPase profile resulted from the treatment, but a complete myosin isoenzymic transition was observed. These results tend to indicate that a moderate increase in the level of thyroid hormone is sufficient to induce the differentiation of adult fiber types, together with the production of adult myosin isoforms in the skeletal dorsal muscle of amphibians, while a pronounced increase would be necessary for repressing the initial larval features.

Analysis of MyoD, myogenin, and muscle-specific gene mRNAs in regenerating Xenopus skeletal muscle

We have analyzed in adult Xenopus laevis, using in situ hybridization, the spatial and temporal expression patterns of MyoD, myogenin, and α‐skeletal actin and fast myosin heavy chain mRNAs during muscle regeneration following cardiotoxin injury. MyoD transcripts could be detected in the satellite cells as early as the first stage of regeneration and were expressed persistently throughout the regeneration process. Myogenin mRNAs were transiently expressed in forming myotubes. α‐Skeletal actin and fast myosin heavy chain mRNAs were detected precociously, before the young myotube stage. This work has shown, for the first time, the presence of myogenin transcripts during Xenopus myogenesis.

Thyroidal status and myosin isoenzymic pattern in the skeletal dorsal muscle of urodelan amphibians — The perennibranchiate Proteus anguinus

In the perennibranchiate Proteus anguinus, larval myosin isoforms were shown to coexist for life with the adult isomyosins that appeared at the end of the larval stage. Analysis of the myofibrillar ATPase profile also revealed that a high percentage of immature fibers persisted in adults. A long-term treatment with large amounts of T3 had no effect on juvenile individuals. Applied to subadult animals it promoted a regression of larval myosin isoforms and a reduction in the percentage of immature fiber types. The regulative effect of T3 in the myosin isoenzymic transition may be delayed and depends on metabolic conditions, which suggests it is indirect.

Changes in calmodulin level after fertilization and during first cleavage in the egg of the urodelan amphibian Pleurodeles waltlii

We report three significant calmodulin rises related to Pleurodeles waltlii egg fertilization and following developmental events. These elevations are correlated to the major obvious Ca2+-dependent events: Na+-H+ exchange, activation of NAD kinase, triggering of cortical reaction, resumption of meiotic division II, initiation of DNA synthesis and regulation of cell division. Therefore, it is suggested that alterations in calmodulin level in fertilized egg may be part of the Ca2+-dependent regulatory mechanisms which turn on metabolisms, initiate development and govern cell cleavages.

Myosin isoforms and their light chains from the ventricular muscle of the urodelan amphibian Pleurodeles waltlii: Comparison with myosin from skeletal muscles☆

Myosin extracted from ventricular muscle of the urodelan amphibian Pleurodeles waltlii was analyzed in comparison with myosin extracted from skeletal muscles by native, one-dimensional SDS gel electrophoresis and two-dimensional gel electrophoresis. Two myosin isoforms were detected in ventricular muscle using pyrophosphate gel electrophoresis. These isomyosins contained two types of light chain subunits, LC1v and LC2v. Two-dimensional gel electrophoresis showed that LC1v comigrated with the slow light chain LC1s, whereas LC2v was characterized by a specific mobility, distinct from LC2s and LC2f. Diaphragm muscle was characterized by the coexistence of larval and adult myosin isoforms.

ANALYSIS OF MUSCLE ADAPTATIONS TO TERRESTRIAL STEPPING IN THE URODELAN AMPHIBIAN PLEURODELES WALTLII

Abstractu2002The changes of myosin isoform pattern and of its associated light chains in relation to the myosin ATPase profile were analysed in different muscles of the hypothyroidian amphibian Pleurodeles waltlii submitted to terrestrial stepping, using electrophoretic and histochemical techniques. These changes were specific to the muscle type but appeared globally characterized by a type-IIB to type-IIA/I fibre transition associated with a transition from fast to intermediate and/or slow myosin isoforms. These results are similar to the effects of endurance training on locomotor muscles of mammals. The diaphragm of experimental animals was also characterized by a complete disappearance of the larval myosin isoforms which were detected in the diaphragm of control animals. The myosin pattern of ventricular muscle did not change following terrestrial stepping. This work indicates that thyroid hormone does not regulate the muscle adaptations that occur following terrestrial stepping and suggests a more complex mechanism of regulation in which innervation could be implicated.