Chantal Janmot

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.

Myosin isoform transitions in regeneration of fast and slow muscles during postnatal development of the rat

Regeneration of rat fast (gastrocnemius medialis) and slow (soleus) muscles was examined after degeneration of myofibers had been achieved by injection of cardiotoxin into the hindleg during the first week after birth. Myogenesis in the regenerating muscles was compared to postnatal myogenesis in the contralateral and in control muscles. Synthesis of embryonic and neonatal myosin isoforms was initiated 3 days after injury. These forms were gradually replaced by the intermediate and fast adult isoforms (type II fiber myosins), whose synthesis followed the same curve in regenerating, contralateral, and control muscles. In contrast, synthesis of the slow myosin isoform (type I fiber myosin) was greatly delayed in injured muscles, but eventually became equal to its synthesis in contralateral and control muscles. It therefore appears that synthesis of type II fiber myosins is similarly regulated, probably by thyroid hormone, in developing regenerating and normal muscles, while synthesis of type I fiber myosin depends on other factor(s).

Electrophoretic separation of developmental and adult rabbit skeletal muscle myosin heavy chain isoforms: Example of application to muscle denervation study

We present the separation by SDS gel‐electrophoresis of the six main myosin heavy chains (MHC) present in rabbit skeletal muscle. The separation of the four adult MHC (1, 2A, 2X/2D, 2B) was compared to that of the corresponding rat MHC as described by Talmadge and Roy [J. Appl. Physiol. 99 (1993) 2337‐2340]. We found that many rabbit muscles contained mainly one of the four MHC, in some cases the 2B MHC. In addition, we resolved the embryonic E and perinatal P developmental MHC, which should facilitate muscle differentiation and regeneration studies in the rabbit. An example of application to the study of muscle denervation is given.

Myosin switches in skeletal muscle development of an urodelan amphibian, Pleurodeles waltlii. Comparison with a mammalian, Mus musculus

The isomyosins from dorsal axial muscle, which appear successively through metamorphosis of P.waltlii, are shown to be composed of identical fast-type light chains but of distinct heavy subunits. We observe that this modification goes with a change in ATPase activity as also in the case of mouse. Metamorphosis in amphibian as well as birth in mammalian are thus both accompanied by the synthesis of new myosins of higher catalytic efficiency.

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.

Myosin Heavy Chain Profiles in Regenerated Fast and Slow Muscles Innervated by the Same Motor Nerve Become Nearly Identical

Plasticity of mature muscles exposed to different activation patterns is limited, probably due to restricted adaptive range of their muscle fibres. In this study, we tested whether satellite cells derived from slow muscles can give rise to a normal fast muscle, if transplanted to the fast muscle bed. Marcaine-treated rat soleus and extensor digitorum longus (EDL) muscles were transplanted to the EDL muscle bed and innervated by the ‘EDL’ nerve. Six months later expression of myosin heavy chain isoforms was analysed by areal densities of fibres, binding specific monoclonal antibodies, and by SDS gel electrophoresis. Both regenerated muscles closely resembled each other. Their myosin heavy chain profiles were similar to those in fast muscles although they were not identical to that in the control EDL muscle. Since not even regenerated EDL was able to reach the myosin heavy chain isoform profile of mature EDL muscle, our experimental model did not permit studying the adaptive capacity of satellite cells in different muscles in its whole extent. However, the results favour the multipotential myoblast stem cell population in rat muscles and underline the importance of the extrinsic regulation of muscle phenotype.

Molecular characteristics of cocaine-induced cardiomyopathy in rats

Cocaine abuse induces severe cardiomyopathy. To investigate the molecular effects of acute and prolonged administration of cocaine, mRNAs encoding markers of either mechanical overload, as atrial natriuretic factor (ANF) and alpha- and beta-myosin heavy chains, or fibrosis as type I and III procollagens, were quantitated in the left ventricle of rats 4 h after one injection of cocaine (40 mg/kg, n = 7), or 14 (n = 15) and 28 days (n = 10) after chronic infusion of cocaine (40 mg/kg per day). Plasma cocaine and benzylecgonine concentrations were both significantly augmented during the infusion while plasma levels of triiodothyronine and thyroxine were lowered. Acute injection of cocaine induced ANF gene expression. Cocaine treatment during 28 days resulted in left ventricular hypertrophy (+ 20% after 24 days, P < 0.05) with normal blood pressure, associated with an accumulation of mRNAs encoding ANF and type I and III collagens (+66% and +55%, P < 0.05). Such a chronic treatment also induced a shift from the alpha- to the beta-myosin heavy chain gene expression (-40% and +50%, P < 0.05). In conclusion, cocaine activates markers of both hemodynamic overload and fibrosis. Such an activation may result from direct and/or indirect effects of the drug such as myocardial ischemia, mechanical overload and/or hypothyroidism.

Myosin isoforms in mackerel (Scomber scombrus) red and white muscles

Myosin extracts of red and white muscles from mackerel (Scomber scombrus) were analyzed by native electrophoresis to investigate the existence of myosin isoforms in both kind of muscles, and by two‐dimensional and SDS gel electrophoresis to study their subunit composition. Two isoforms were found in red muscle comprising one type of heavy chain and two light chains. Four isoforms were found in white muscle made up of one type of heavy chain and three types of light chains. The heavy chains from white muscle showed a higher electrophoretic mobility than that of red muscle in SDS‐PAGE. Both heavy chains had an intermediate mobility between those of slow and fast myosins from rat diaphragm.

Opposite regulations by androgenic and thyroid hormones of V1 myosin expression in the two types of rabbit striated muscle: skeletal and cardiac.

The finding that VI cardiac myosin is expressed in masticatory skeletal muscles of the rabbit [1] provided a unique opportunity for comparing the hormonal regulation of VI in skeletal and cardiac muscles. Thyroid hormones had no significant effect on the postnatal expression of VI in masticatory muscles, but increased this expression in cardiac ventricles. In contrast, androgenic hormones reduced VI expression in masticatory muscles, but did not affect it significantly in cardiac ventricles. Modulation of VI gene transcription in striated muscle is thus shown here to depend both on the target muscle and on the hormone.

Reinnervation of denervated extensor digitorum longus of the rat by the nerve of the soleus does not induce the type I myosin synthesis directly but through a sequential transition of type II myosin isoforms

The fast-contracting extensor digitorum longus (EDL) muscle of 1-month-old rats was denervated and reinnervated by the nerve innervating the slow-contracting soleus muscle. After variable periods of time, the myosin isoform content of the EDL was analyzed by sensitive electrophoretic techniques, which allowed to discriminate between the slow-type I and the three, IIA, (IID or IIX) and IIB, fast-type II myosin isoforms. Compared to the control EDL, which contains predominantly the IIB isoform, the operated muscles contained variable proportions of all the isoforms. Analysis of the results leads us to conclude that reinnervation of EDL induces a sequential transition of myosin isoforms: IIB----(IID or IIX)----IIA----I.