Coexistence of slow and fast isoforms of contractile and regulatory proteins in human skeletal muscle fibres induced by endurance raining

The distribution of fast and slow isoforms of troponin C, I, and T components and myosin heavy chains was investigated in histochemically typed myofibrillar ATPase intermediate (IM) fibres, that is, fibres that stain after both acid and alkaline pre-incubation in stainings for myofibrillar ATPase. In addition to the previously described IM fibres of types II C and I B, fibres that displayed staining characteristics between types II C and I B were observed and termed type II C–I B. The IM fibres constitute less than 1% of the fibres in normal human limb and abdominal muscles. The IM fibres studied here resulted from extensive endurance training of human triceps brachii muscle (n= 6) and were induced by conversion of a proportion (130) of type II fibres. The immunohistochemical stains of serial sections with antibodies to slow isoforms of troponin I, T, C and myosin heavy chain showed no staining of type II fibres but intense staining of types I and I B fibres, whereas type II C fibres stained with intermediate intensity. The antibodies to fast isoforms of the troponin components and myosin heavy chain did not give rise to staining of type I fibres but dark staining of type II fibres. Type I B fibres stained with intermediate intensity and type II C was either as dark as type II or slightly lighter. Type II C-I B fibres showed staining intensities intermediate between those observed for types I B and IIC in the immunohistochemical stains. It is therefore concluded that training-induced myofibrillar ATPase intermediate human skeletal muscle fibres are characterized by the coexistence of slow and fast isoforms of contractile and regulatory proteins. Changes in the distribution of fast and slow isoforms of several of the myofibrillar proteins appeared to be induced in a co-ordinated manner.     

Cortisone-induced changes in myosin heavy chain distribution in respiratory and hindlimb muscles

In this study the effects of administration of cortisone acetate (100 mg kg^-1 body weight subcutaneously for 11 days) on distribution and cross-sectional area of different fibre types of rat skeletal muscles were investigated. Diaphragm, parasternal intercostal (PI), extensor digitorum longus (EDL) and soleus muscles were examined in cortisone treated animals (CA) in comparison with ad libitum controls (CTRL) and pair-fed (PF) controls. Four fibre types (I or slow and IIA, IIX, IIB or fast) were identified on the basis of their myosin heavy chain composition using a set of monoclonal antibodies. In CA rats the reduction of cross-sectional area was above 30% in IIX fibres of diaphragm, IIB fibres of PI and in all fast fibres of EDL. In all muscles slow fibres were spared from atrophy. Significant variations in fibre type distribution were found in the muscles of CA rats when compared to CTRL. The percentage of IIB fibres decreased in EDL, PI and diaphragm. This decrease was accompanied by an increase in the percentage of IIA fibres in the same muscles. No changes in the percentage of slow fibres and of fast IIX fibres were observed in EDL, PI and diaphragm of CA rats in comparison with CTRL. In soleus of CA rats the proportion of IIA fibres was lower than in CTRL. In EDL of PF rats atrophy of IIA fibres and changes in fibre type distribution were similar to those observed in CA rats. In diaphragm, PI and soleus of PF rats no significant decrease in fibre cross-sectional area nor significant changes in fibre distribution were found in comparison with CTRL rats.     

Functional characterization of the human atrial essential myosin light chain (hALC-1) in a transgenic rat model

Most patients with hypertrophic cardiomyopathy and congenital heart diseases express the atrial essential myosin light chains (ALC-1) in their ventricles, partially replacing the ventricular essential light chains (VLC-1). This VLC-1/ALC-1 isoform shift is correlated with an increase in cross-bridge cycling kinetics as measured using skinned fibers from the hypertrophied ventricles of human hearts.To study the functional importance of hALC-1 in the intact perfused heart, we generated a transgenic rat model (TGR) overexpressing hALC-1 in the heart. Twelve-week-old TGR rats expressed 17±4 g hALC-1 per mg of whole SDS-soluble protein. Their perfused heart contractility parameters were evaluated using the Langendorff preparation. Expression of hALC-1 was accompanied by statistically significant improvements (P<0.001) in the contractile parameters of the hearts of the TGR compared to the age matched control (WKY) animals, represented by increases from 20.8±2.3 to 45.1±3.6 mmHg/g heart weight in the developed left ventricular pressure, 1,035.7±89.8 to 2,181±135.4 mmHg/s in the contraction rate, and 713±60.2 to 1,364±137.4 mmHg/s in the relaxation rate in the WKY and the TGR groups respectively. Characterizing the functional effects of hALC-1 at the whole organ level represents a step towards gene therapy of heart failure.     

Myosin heavy chain expression and atrophy in rat skeletal muscle during transition from cardiac hypertrophy to heart failure

The purpose of this investigation was to determine whether changes in myosin heavy chain (MHC) expression and atrophy in rat skeletal muscle are observed during transition from cardiac hypertrophy to chronic heart failure (CHF) induced by aortic stenosis (AS). AS and control animals were studied 12 and 18 weeks after surgery and when overt CHF had developed in AS animals, 28 weeks after the surgery. The following parameters were studied in the soleus muscle: muscle atrophy index (soleus weight/body weight), muscle fibre diameter and frequency and MHC expression. AS animals presented decreases in both MHC1 and type I fibres and increases in both MHC2a and type IIa fibres during late cardiac hypertrophy and CHF. Type IIa fibre atrophy occurred during CHF. In conclusion, our data demonstrate that skeletal muscle phenotype changes occur in both late cardiac hypertrophy and heart failure; this suggests that attention should be given to the fact that skeletal muscle phenotype changes occur prior to overt heart failure symptoms.     

Correlation between percentage fiber type area and myosin heavy chain content in human skeletal muscle

Histochemical methods are routinely used to delineate skeletal muscle fiber types. In the present investigation, this qualitative determination of fiber type composition was compared to the electrophoretically determined myosin heavy chain (MHC) content from a large number of human muscle biopsy samples. Biopsies were taken from the vastus lateralis muscle at the beginning and every 2 weeks during 8 weeks of highi-ntensity resistance training from men (n = 13) and woman (n = 8). Muscle was also extracted from nontraining men (n = 7) and women (n = 5) at the same periods. Six muscle fiber types (I, IC, IIAC, IIA, IIAB, and IIB) were determined using basic myofibrillar adenosine triphosphatase histochemistry. Cross-sectional areas were determined for the three major fiber types (I, IIA, and IIB) and used to calculate the percentage area of these types. Electrophoretic techniques were used to separate and quantify the percentage MHC content in these same biopsy samples, and these data were then used to compare with the percentage fiber type area. Correlation analyses suggest a relationship between the histochemically assessed percentage fiber type area and the electrophoretically assessed MHC content in human limb musculature. However, because of possible histochemical misclassification of some fibers (especially in trained muscle) both techniques may be essential in yielding important information about fiber type composition and possible fiber type transformations.     

Influence of fast and slow alkali myosin light chain isoforms on the kinetics of stretch-induced force transients of fast-twitch type IIA fibres of rat

This study contributes to understand the physiological role of slow myosin light chain isoforms in fast-twitch type IIA fibres of skeletal muscle. These isoforms are often attached to the myosin necks of rat type IIA fibres, whereby the slow alkali myosin light chain isoform MLC1s is much more frequent and abundant than the slow regulatory myosin light chain isoform MLC2s. In the present study, single-skinned rat type IIA fibres were maximally Ca²⁺ activated and subjected to stepwise stretches for causing a perturbation of myosin head pulling cycles. From the time course of the resulting force transients, myosin head kinetics was deduced. Fibres containing MLC1s exhibited slower kinetics independently of the presence or absence of MLC2s. At the maximal MLC1s concentration of about 75%, the slowing was about 40%. The slowing effect of MLC1s is possibly due to differences in the myosin heavy chain binding sites of the fast and slow alkali MLC isoforms, which changes the rigidity of the myosin neck. Compared with the impact of myosin heavy chain isoforms in various fast-twitch fibre types, the influence of MLC1s on myosin head kinetics of type IIA fibres is much smaller. In conclusion, the physiological role of fast and slow MLC isoforms in type IIA fibres is a fine-tuning of the myosin head kinetics.     

Force-velocity properties and myosin light chain isoform composition of an identified type of skinned fibres from rat skeletal muscle

Force-velocity relations, myosin heavy chain (MHC) and myosin light chain (MLC) isoform composition of single skinned fibres from rat plantaris muscle were determined. In fibres containing the same (2X) isoform of myosin heavy chain, several parameters derived from the force-velocity relation and isometric force (Po) were tested for relation with the fibre content in alkali myosin light chain (MLC) isoforms. Whereas maximum shortening velocity was found to be proportional to the relative content in the 3f isoform of alkali MLC, velocity of shortening at 5% relative load, maximum power output, and Po were not. These results strengthen the idea that, in mammalian skeletal fibres, alkali MLC isoforms modulate shortening velocity at zero load, but suggest that they do not control the contractile behaviour at loads higher than zero.     

Myosin heavy chain isoform transition in ageing fast and slow muscles of the rat

Sugiura , T., Matoba , H., Miyata , H., Kawai , Y. & Murakami , N. 1992. Myosin heavy chain isoform transition in ageing fast and slow muscles of the rat. Acta Physiol Scand 144 , 419423. Received 26 August 1 991 , accepted 3 December 1991. ISSN 0001–6772. Laboratory of Biomechanics and Physiology, Faculty of Liberal Arts, Yamaguchi University, Yamaguchi, Faculty of Integrated Arts and Sciences, Tokushima University, Tokushima, and Department of Physiology, Yamaguchi University School of Medicine, Ube, Yamaguchi, Japan. Using gradient sodium dodecyl sulphate-polyacrylamide gel electrophoresis, myosin heavy chain (MHC) isoforms were studied in the extensor digitorum longus (EDI,) and the soleus muscles of male Wistar rats at different ages ( 5, 10 , 20 weeks, 1 and 2 years). In the EDL muscle, four types of MHC isoforms were observed in all age groups. There was an increase in the percentage of HCIId and a concomitant decrease in the percentage of HCIIb with increasing age. No significant difference was observed in the percentages of HCI and HCIIa isoforms in all the age groups. In contrast, the soleus muscle contained two MHC isoforms, HCI and HCIIa. There was an increase in the percentage of HCI and a concomitant decrease in the percentage of HCIIa with increasing age. These results suggest that age-related changes in the MHC isoforms in both the fast-twitch EDI, and the slow-twitch soleus muscles are one factor underlying the age-related decrease in the speed of muscle contraction.     

Effects of low and high frequency patterns of stimulation on contractile properties,enzyme activities and myosin light chain accumulation in slow and fast denervated muscles of the chicken

The effects of denervation and direct stimulation in fast and slow latissimus dorsii muscles were investigated in chicken. In slow ALD muscle, denervation resulted in an incompleteness of the relaxation, a decrease in MDH and CPK activities and an increase in fast myosin light chains (MLC) accumulation. Direct stimulation at either fast or slow rhythm prevented the effects of denervation on relaxation and CPK activity but was ineffective on MDH activity and fast MLC accumulation. Moreover, direct stimulation of denervated ALD caused rhythm-dependent change in tetanic contraction. In fast PLD muscle, the main changes in muscle properties following denervation were a slowing down of the time course of the twitch and an incompleteness of the relaxation, a decrease in LDH and CPK activities and in LC₃F accumulation. Stimulation at a high frequency partly prevented the effects of denervation and resulted in a large accumulation of LC₃F, while a low frequency stimulation did not restore the twitch time to peak, increased MDH activity and induced synthesis of slow MLC. This study emphasizes the role of muscle activity and its pattern in some properties of slow and fast chicken muscles following denervation.This work was supported by the Association des Myopathes de France     

Mechanical properties and myosin heavy chain isoform composition of skinned skeletal muscle fibres from a human biopsy sample

 Experiments were conducted to investigate the mechanics of contraction of chemically skinned muscle fibre segments of a biopsied sample of single human quadriceps muscle. Subsequently, the isoforms of the myosin heavy chain (MHC) were analysed by sodium dodecyl sulphate (SDS) gel electrophoresis.Of the 41 fibres, 26 contained MHCI (type I), 11 of the fibres contained MHCIIa (type IIA), and 4 of the fibres contained both MHCI and MHCIIa (of which MHCIIa was always slightly predominant (type IIC)). Distinct differences between fibre types were found in terms of the kinetics of force responses following stepwise length changes (order of velocity: IIA > IIC > I). The differences in maximal shortening velocity and in the kinetics of Ca²⁺-dependent activation were of the same order, but much less pronounced. Type I fibres had significantly greater fibre diameters than type IIA fibres. No significant differences were found among different fibre types in terms of isometric tension, resting sarcomere length or the length change needed to discharge the elasticity of maximally Ca²⁺-activated fibres (y _o value). The distribution of shortening velocity and kinetics of stretch activation values suggest that two muscle fibre subtypes may exist in human type I fibres. Received: 15 April 1997 / Received after revision 19 May 1997 / Accepted: 20 May 1997     

Detection of complement C3 and factor B gene expression in normal colorectal mucosa,adenomas and carcinomas

Local secretion of complement components in the human intestine has been previously reported. However, the cellular source has not been identified. In this study, we demonstrate complement C3 and factor B mRNA expression in the normal colonic mucosa by in situ hybridization analysis. C3 and factor B genes were found to be expressed at high levels in the epithelial cells of the lower parts of the crypts in colonic mucosa, and this expression decreased gradually from the crypt base to the luminal surface. At the upper crypt and the luminal surface, these genes almost disappeared. C3 and factor B genes were expressed in all crypts at the same level. Furthermore, C3 and factor B gene expression was also identified in adenomas and carcinomas. In these neoplastic tissues, C3 and factor B genes were expressed uniformly, and the polarized distribution observed in the normal crypts was not detected. It is likely that complement components are locally synthesized in the intestine, and that these complement components may actively participate in normal immune and inflammatory responses over the enormous surface area of the intestinal mucosa.     

Smooth muscle alpha actin and myosin heavy chain expression in the vascular smooth muscle cells surrounding human endometrial arterioles

Endometrial spiral arterioles are believed to play a major role in controlling menstruation. These arterioles coil and grow through the secretory stages of the cycle, unlike the 'straight' endometrial arterioles that remain uncoiled. We postulate that alterations in the growth and development of spiral arterioles, in particular the vascular smooth muscle cells (VSMC), may contribute to menorrhagia. We examined smooth muscle alpha actin (alphaSMA) and myosin heavy chains (MHC), two VSMC differentiation markers, in the endometrial arterioles of 64 women, comparing them in controls, menorrhagic tissues and across the menstrual cycle. alphaSMA and MHC expression were determined immunohistochemically then evaluated using computer-aided image analysis. alphaSMA expression in the straight arterioles of menorrhagic women was reduced in the early secretory stage of the cycle and significantly decreased at the mid-secretory stage of the cycle (0.67 +/- 0.03 versus 0.55 +/- 0.04, P 相似文献   

Time dependent effects on contractile properties,fibre population,myosin light chains and enzymes of energy metabolism in intermittently and continuously stimulated fast twitch muscles of the rabbit

Summary Fast-twitch tibialis anterior and extensor digitorum longus rabbit muscles were subjected to long-term intermittent (8 h daily) or continuous (24 h daily) indirect stimulation with a frequency pattern resembling that of a slow motoneuron.Increases in time to peak of isometric twitch contraction were observed without parallel changes in the pattern of myosin light chains or alterations in the distribution of slow and fast fibres as discernible by the histochemical ATPase reaction. However, changes in the fibre population and in the myosin light chain pattern were observed after intermittent stimulation periods exceeding 40 days or continuous stimulation periods longer than 20 days. Under these conditions even higher increases were found in contraction time. In one animal a complete change in fibre population was observed. In this case myosin light chains of the slow (LC_S1, LC_S2) and of the fast type (LC_f1) were obviously synthetized simultaneously within the same fibre. Early changes in the enzyme activity pattern of energy metabolism indicated a conversion of the fibres including their mitochondrial population. These changes and the earlier reported changes in the sarcoplasmic reticulum are probably responsible for the early changes in contractile properties which occur before the transformation of the myosin.This study was supported by Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 138 Biologische Grenzflächen und Spezifität and by grants from Deutscher Sportbund     

Myosin heavy chains in skeletal muscles of the common shrew (Sorex araneus): absence of a slow isoform and transitions of fast isoforms with ageing

Myosin heavy chain (MHC) composition in seven skeletal muscles of the common shrew (Sorex araneus) was analysed by gradient SDS-PAGE and immunoblotting with monoclonal antibodies. Characteristic for the studied muscles (diaphragm, lateral gastrocnemius, medial gastrocnemius, masseter, plantaris, soleus and tibialis anterior) was a total absence of the slow isoform, MHCI; all muscles were exclusively composed of two fast isoforms MHCIId and MHCIIb. In young adults the amount of MHCIId varied between 34% (tibialis anterior) and 97% (masseter). In over-wintered senescent individuals MHCIId was clearly the dominant isoform in all muscles studied; the lowest MHCIId content was measured for tibialis anterior (69%), while in diaphragm, masseter and soleus it was practically the sole isoform (over 96%). Ageing associated isoform transition from MHCIIb to MHCIId occurred in all seven muscles studied (P< 0.05).     

Store and recoil of elastic energy in slow and fast types of human skeletal muscles

Stretch-shortening cycle refers to the mechanical condition in which store and recoil of elastic energy occur in the skeletal muscle. This leads to a greater work output when compared to a simple shortening contraction. The subjects performed vertical jumps with and without preliminary counter-movement and with small and large knee angular displacement. The results indicated that those subjects who had more fast twitch (FT) fibers benefited more from the stretching phase performed with high speed and short angular displacement. The amounts of elastic energy stored in this phase were 30 and 26 N × kgBW^-1, respectively, for FT and slow twitch (ST) type subjects. The recoil of elastic energy was proportional to the amount of energy storage. In large amplitude jumps where transient period between stretch and shortening is long the both types of subjects demonstrated similar amount of storage of elastic energy (17 and 16 N × kgBW^-1, respectively). However, the re-use of this elastic energy was greater in ST group (24%) as compared to the FT group (17%). The results can be interpreted through differences in sarcomere cross-bridge life times between fast and slow muscle fibers. The slow type muscle may be able to retain the cross-bridge attachment for a longer period of time and therefore it may utilize elastic energy better in a slow type ballistic motion.     

Nonuniform myosin expression along single fibers of chronically stimulated and contralateral rabbit tibialis anterior muscles

Using a combined histochemical and biochemical technique, single fiber analyses were performed on chronically stimulated and contralateral tibialis anterior (TA) muscles of the rabbit. The major fiber population (60%) in 30 days stimulated TA was transforming fibers (type IIC). Some of these fibers displayed a nonuniform distribution of histochemically assessed myofibrillar actomyosin ATPase (mATPase) activity. This heterogeneity of mATPase activity along the fibers was verified in longitudinal sections and by microphotometric evaluation of mATPase staining intensities in serial cross-sections. Biochemical analyses of single fiber segments revealed that these C fibers not only coexpressed fast- and slow-myosin subunits but did so nonuniformly along their length. The distribution of fast- and slow-myosin subunits in these fibers was not random but focal. Variations in myosin expression were also observed in some of the C fibers in the contralateral TA. As opposed to the transforming fibers in the stimulated TA, heterogeneities of mATPase activity and myosin subunits in these contralateral C fibers were less focal and more gradual. These findings suggest that muscle fibers in chronically stimulated TA and the contralateral muscle do not transform synchronously or uniformly along their length.     

Exercise-induced fibre type transitions with regard to myosin,parvalbumin, and sarcoplasmic reticulum in muscles of the rat

Effects of a long-term, high intensity training program upon histochemically assessed myofibrillar actomyosin ATPase, myosin composition, peptide pattern of sarcoplasmic reticulum (SR), and parvalbumin content were analysed in muscles from the same rats which were used in a previous study (Green et al. 1983). Following 15 weeks of extreme training, an increase in type I and type IIA fibres and a decrease in type IIB fibres occurred both in plantaris and extensor digitorum longus (EDL) muscles. In the deep portion of vastus lateralis (VLD), there was a pronounced increase from 10±5% to 27±11% in type I fibres. No type I fibres were detected in the superficial portion of vastus lateralis (VLS) both in control and trained animals. An increase in slow type myosin light chains accompanied the histochemically observed fibre type transition in VLD. Changes in the peptide pattern of SR occurred both in VLS and VLD and suggested a complete transition from type IIB to IIA in VLS and from type IIA to I in VLD. A complete type IIA to I transition in the VLD was also suggested by the failure to detect parvalbumin in this muscle after 15 weeks of training. Changes in parvalbumin content and SR tended to precede the transitions in the myosin light chains. Obviously, high intensity endurance training is capable of transforming specific characteristics of muscle fibres beyond the commonly observed changes in the enzyme activity pattern of energy metabolism. The time courses of the various changes which are similar to those in chronic nerve stimulation experiments, indicate that various functional systems of the muscle fibre do not change simultaneously.     

The effect of different patterns of long-term stimulation on contractile properties and myosin light chains in rabbit fast muscles

Fast rabbit skeletal muscles (tibialis anterior and extensor digitorum longus) were stimulated for 2–28 days by electrodes implanted in the vicinity of the peroneal nerve to produce maximal contractions at two different frequency patterns: that occuring naturally in nerves to slow muscles (10 Hz continuously) or three bursts of tetani (40 Hz) per minute, each 5s in duration. Both types of frequency produced muscles more resistant to fatigue during isometric twitch contractions, and led to a prolongation of contraction time greater and more consistent with 10 Hz than with 40 Hz. The twitch/tetanus ration was significantly higher in muscles stimulated at 10 Hz for 3–4 weeks but was not different from controls in muscles stimulated at 40 Hz. Both types of stimulation led to the appearance of myosin light chains characteristic of slow muscles. Muscles stimulated for 4 weeks at 40 Hz developed greater twitch tension per gram, and had significantly smaller cross-sectional area of myofibrils than control muscles. It is concluded that long-term electrical stimulation of fast muscles can affect some muscle contractile properties to resemble those of slow muscles irrespective of frequency of stimulation, provided the total number of stimuli is comparable, the duration of stimulation is long enough (minimum 2 weeks) and all motor units are activated.This work was supported by a grant from British Medical Research Council, and from Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 138 Biologische Grenzflächen und Spezifität