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Biomedical subjects

A Margreth

Publications and source records attributed to A Margreth.

At least 73 records · Page 4Linked to original sources

Evidence for the presence of the stearyl-CoA desaturase system in the sarcoplasmic reticulum of rabbit slow muscle.

We have shown that the isolated sarcoplasmic reticulum from rabbit slow muscle contains cytochrome b5 which can be reduced via a flavoprotein, with FAD as the prosthetic group. In the presence of NADH and oxygen, these sarcoplasmic reticulum membranes can convert stearyl-CoA to oleyl-CoA, similarly to liver endoplasmic reticulum membranes. However, the stearyl-CoA desaturase system is virtually lacking in fast muscle sarcoplasmic reticulum. The data suggest that these differences between fast and slow twitch muscle may be related to the characteristic fatty acid composition of phospholipids and the function of the sarcoplasmic reticulum.

Animals↗

Microplate enzyme-linked immunosorbent assay in the study of the structural relationship between myosin light chains.

A microplate enzyme-linked immunosorbent assay (microELISA) for the study of immunochemical relationships between rabbit myosin light chains is described. Purified individual fast-muscle myosin light chains (LC1F, LC2F and LC3F) and their respective antisera, obtained in chicken, were used. Optimal conditions for antigen concentration, antiserum dilution, substrate concentration, incubation time and reproducibility with time were established. The observed cross-reactivities between the different types of light chains associated with rabbit fast-muscle myosin confirm and extend previous results obtained by other authors using radioimmunoassay procedures. It was concluded that microELIAS may be successfully employed also to the study of macromolecule cross-reactivities.

Animals↗

[Muscular phenotypes in relation to the specific differentiative influences of motor innervation].

Vertebrate skeletal muscles are classified into fast-twitch and slow-twitch muscles according to the intrinsic speed of contraction. These physiological characteristics of the muscles are ontogenetically determined by epigenetic influences arising from the specific motor innervation. The evidence for a neural control on gene expression comes mainly from the demonstration that myosin is present in two different molecular forms in fast and in slow muscles. It is known from previous work in several laboratories that the two myosin isozymes differ with respect to both the primary structure of the heavy chains and the subunit composition of the light chains. Fast muscle myosin is characterized by a three-bands electrophoretic pattern in SDS-gels, whereas the myosin from slow muscle contains only two, distinct types of subunits. Our results show that the tripartite band pattern of the light chains is a common characteristic for the myosin of the fast-white muscles with intermittent-phasic activity (e.g. rabbit adductor) and the fast-red muscles with sustained-phasic activity (e.g. rat masseter and pigeon pectoralis). These results lend support to the view that the neural control on gene expression in skeletal muscles is mediated by specific influences somehow arising from the pattern of activity and which are independent from the total input of nerve impulses.

Animals↗

Structural membrane proteins and loosely associated proteins of the sarcoplasmic reticulum.

The protein composition of sarcoplasmic-reticulum vesicles, either unpurified or after fractionation on sucrose gradients, and with or without previous osmotic shock and sonication, was investigated by electrophoresis in acid polyacrylamide gels. The pattern of release of loosely bound proteins is discussed with respect to their localization in the interior of the vesicles.

Adenosine Triphosphatases↗

Early biochemical consequences of denervation in fast and slow skeletal muscles and their relationship to neural control over muscle differentiation.

1. One week after denervation several biochemical characteristics of the fast extensor digitorum longus and slow soleus muscles from adult rats were investigated and compared with the characteristics of the corresponding unoperated contralateral muscles. 2. After these short periods of denervation-induced atrophy, the isolated myosins showed unchanged ATPase (adenosine triphosphatase) activities, but there was the expected difference between fast and slow muscle. 3. The specific activities of several soluble enzymes and their characteristic patterns were found to be only slightly modified in both the extensor and soleus muscles after denervation, as were most of the activities measured in the isolated mitochondria. 4. The most significant modifications were in the isolated sarcoplasmic reticulum, and appeared to be specific to either slow or fast muscle. 5. Denervation of slow muscle led to a marked increase of Ca(2+)-transport rates, and of the specific activity of the Mg(2+)-activated K(+)-modulated Ca(2+)-stimulated ATPase, together with changes in the polyacrylamide-electrophoretic profiles of the microsomal membrane protein. Transformation of these several properties of slow muscle sarcoplasmic reticulum to those of fast muscle sarcoplasmic reticulum was further substantiated by electron-microscopic analysis after negative staining. Control experiments with tenotomized soleus muscle gave negative results. 6. The isolated sarcoplasmic reticulum from fast muscle showed a slight diminution of ATPase-linked Ca(2+)-transport activity and a selective increase of rotenone-insensitive NADH-cytochrome c reductase activity, in addition to a greater emphasis on slow-type electrophoretic components of the structural membrane protein. 7. The significance of these results in relation to specific differentiating influences from motor nerves is discussed.

Adenosine Triphosphatases↗

Coordinated development of the sarcoplasmic reticulum and T system during postnatal differentiation of rat skeletal muscle.

An electron microscope study has been carried out on rat psoas muscle, during the early postnatal stages of development. Among the several subcellular components, the sarcotubular system undergoes the most striking modifications during this period. In muscle fibers of the newborn rat, junctional contacts between the T system and the SR are sparse and are, mostly, longitudinally or obliquely oriented. The T tubules do not penetrate deeply into the muscle cell, as indicated by the predominantly peripheral location of the triads and the persistence, at these stages of development, of a highly branched subsarcolemmal system of tubules. Diadic associations of junctional SR elements with the plasma membrane are also occasionally observed. The early SR elaborations incompletely delineate the myofibrils, at both the A- and I-band level. Longitudinal sections show irregularly oriented SR tubules, running continuously over successive sarcomeres. Flattened junctional cisterns filled with granular material are sparse and laterally interconnected, at circumscribed sites, with the SR tubules. Between 1 and 2 wk postpartum, transversal triadic contacts are extensively established, at the A-I band level, and the SR network differentiates into two portions in register with the A and I band, respectively. At 10-15 days after birth, the SR provides a transversely continuous double sheet around the myofibrils at the I-band level, whereas it forms a single discontinuous layer at the A-band level. The relationship that these morphological modifications of the sarcotubular system may bear to previously described biochemical and physiological changes of rat muscle fibers after birth is discussed.

Animals↗

On the differential response of sarcoplasm and myoplasm to denervation in frog muscle.

Electron microscopic evidence is presented that the early response to denervation ("simple atrophy") of the semitendinosus m. of the frog is characterized by a greater prominence of the sarcoplasmic reticulum and by the presence, in the interfibrillar spaces, of mitochondria which are more numerous and smaller than in normal muscle. In contrast with the dynamic changes of the sarcoplasmic structural components, the myofibrils showed a progressive decrease in diameter after denervation and throughout the period studied. By carrying out tissue fractionation experiments, the yield of microsome-protein was found significantly greater in the denervated muscles, as compared with the contralateral controls, in this initial stage. Under the conditions attending the overdevelopment of the sarcoplasmic reticulum (SR), denervated semitendinosus m. incorporated valine-C(14) into proteins more actively than the control pairs. The denervated muscles also showed an increase in the number of freely scattered and membrane-bound ribosomes and of polyribosomes, suggesting a more active synthesis of the SR membranes. Pronounced atrophy of the myofibrils, disorganization of the SR, and an increased number of ribonucleoprotein particles lying in the enlarged interfibrillar spaces were the main ultrastructural features of "degenerative atrophy" in frog muscle in the late periods after denervation. The probably adaptive character of the early changes occurring on denervation of frog muscle is discussed.

Animals↗