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E Bandman

Publications and source records attributed to E Bandman.

At least 37 records · Page 2Linked to original sources
PubMed1992

Contractile protein isoforms in muscle development.

The contractile proteins of skeletal muscle are often represented by families of very similar isoforms. Protein isoforms can result from the differential expression of multigene families or from multiple transcripts from a single gene via alternative splicing. In many cases the regulatory mechanisms that determine the accumulation of specific isoforms via alternative splicing or differential gene expression are being unraveled. However, the functional significance of expressing different proteins during muscle development remains a key issue that has not been resolved. It is widely believed that distinct isoforms within a family are uniquely adapted to muscles with different physiological properties, since separate isoform families are often coordinately regulated within functionally distinct muscle fiber types. It is also possible that different isoforms are functionally indistinguishable and represent an inherent genetic redundancy among critically important muscle proteins. The goal of this review is to assess the evidence that muscle proteins which exist as different isoforms in developing and mature skeletal and cardiac muscles are functionally unique. Since regulation of both transcription and alternative splicing within multigene families may also be an important factor determining the accumulation of specific protein isoforms, evidence that genetic regulation rather than protein coding information provides the functional basis of isoform diversity is also examined.

Actins↗
PubMed1992

Skeletal muscle satellite cells appear during late chicken embryogenesis.

The emergence of avian satellite cells during development has been studied using markers that distinguish adult from fetal cells. Previous studies by us have shown that myogenic cultures from fetal (Embryonic Day 10) and adult 12-16 weeks) chicken pectoralis muscle (PM) each regulate expression of the embryonic isoform of fast myosin heavy chain (MHC) differently. In fetal cultures, embryonic MHC is coexpressed with a ventricular MHC in both myocytes (differentiated myoblasts) and myotubes. In contrast, myocytes and newly formed myotubes in adult cultures express ventricular but not embryonic MHC. In the current study, the appearance of myocytes and myotubes which express ventricular but not embryonic MHC was used to determine when adult myoblasts first emerge during avian development. By examining patterns of MHC expression in mass and clonal cultures prepared from embryonic and posthatch chicken skeletal muscle using double-label immunofluorescence with isoform-specific monoclonal antibodies, we show that a significant number of myocytes and myotubes which stain for ventricular but not embryonic MHC are first seen in cultures derived from PM during fetal development (Embryonic Day 18) and comprise the majority, if not all, of the myoblasts present at hatching and beyond. These results suggest that adult type myoblasts become dominant in late embryogenesis. We also show that satellite cell cultures derived from adult slow muscle give results similar to those of cultures derived from adult fast muscle. Cultures derived from Embryonic Day 10 hindlimb form myocytes and myotubes that coexpress ventricular and embryonic MHCs in a manner similar to cells of the Embryonic Day 10 PM. Thus, adult and fetal expression patterns of ventricular and embryonic MHCs are correlated with developmental age but not muscle fiber type.

Animals↗
PubMed1992

Developmental modulation of myosin expression by thyroid hormone in avian skeletal muscle.

It is well established that a rise in circulating thyroid hormone during the second half of chick embryo development significantly influences muscle weight gain and bone growth. We studied thyroid influence on differentiation in slow anterior latissimus dorsi (ALD) and fast posterior latissimus dorsi (PLD) muscles of embryos rendered hypothyroid by hypophysectomy or administration of an anti-thyroid drug. The expression of native myosins and myosin light chains (MLCs) was studied by electrophoretic analysis, and the myosin heavy chain (MHC) was characterized by immunohistochemistry. The first effects of hypothyroid status were observed at day 21 of embryonic development (stage 46 according to Hamburger and Hamilton). Analysis of myosin isoform expression in PLD muscles of hypothyroid embryos showed persistence of slow migrating native myosins and slow MLCs as well as inhibition of neonatal fast MHC expression, indicating retarded differentiation of this muscle. In ALD muscle, hypothyroidism maintained fast embryonic MHC and induced noticeable amounts of fast MLCs, thus delaying slow muscle differentiation. Our results suggest that thyroid hormones play a role in modulating the appearance of neonatal fast MHC and the disappearance of isomyosins transiently present during embryogenesis. However, T3 supplemental treatment would seem to compensate in part for the effects of hypothyroidism induced by hypophysectomy, suggesting that thyroid hormone might interfere with other factors also accounting for the observed effects.

Animals↗
PubMed1991

Myoblasts from fetal and adult skeletal muscle regulate myosin expression differently.

We compared the expression of myosin heavy chains in myogenic cultures prepared from fetal (embryonic Day 10) and adult (12-16 weeks) chicken pectoralis muscle using immunofluorescence with isoform-specific monoclonal antibodies. We found that the majority of fetal myocytes (differentiated myoblasts) and myotubes coexpressed ventricular and embryonic myosin heavy chains in culture. Also, when fetal cells were plated at a clonal density most clones coexpressed both ventricular and embryonic isoforms. In contrast, all adult myocytes and newly formed adult myotubes expressed just ventricular myosin, whether plated at mass or clonal densities. Within 12-24 hr of the onset of fusion, adult myotubes began to express embryonic myosin as well. Eventually, the majority of adult myotubes coexpressed both ventricular and embryonic myosin. The delay of embryonic myosin expression until after fusion was also seen in passaged adult myoblasts and in myoblasts isolated from regenerating adult muscle. The expression of embryonic myosin can be abolished by inhibiting fusion with EGTA in adult but not in fetal cultures. We conclude that both fetal and adult myotubes express ventricular and embryonic myosins but only fetal myocytes express the embryonic isoform prior to fusion. This difference in the regulation of embryonic myosin expression between fetal and adult myoblasts supports the hypothesis that these cells may represent two distinct populations of myogenic precursors.

Aging↗
PubMed1991

Expression of Ca2(+)-ATPase isoforms in denervated, regenerating, and dystrophic chicken skeletal muscle.

The expression of fast and slow isoforms of the sarcoplasmic reticulum Ca2(+)-ATPase was studied in denervated, regenerating, and dystrophic fast and slow avian skeletal muscles. We found that both fast and slow Ca2(+)-ATPase isoforms were expressed in most myofibers following denervation of adult fast-twitch muscle, but only the slow Ca2(+)-ATPase isoform was found in slow-tonic muscle which had been denervated. Regenerating myotubes in normally innervated and previously denervated adult fast-twitch or slow-tonic muscle expressed both Ca2(+)-ATPase isoforms. Expression of the slow Ca2(+)-ATPase isoform was found to persist in dystrophic fast-twitch muscle, long after it had disappeared from normal fast-twitch muscle. However, the fast Ca2(+)-ATPase isoform disappeared from slow-tonic muscle similarly in normal and dystrophic birds. These results demonstrate that the appearance of myosin heavy chain isoforms characteristic of developing muscle is correlated with similar changes in the expression of sarcoplasmic reticulum Ca2(+)-ATPases.

Animals↗
PubMed1991

Neonatal and adult myosin heavy chains form homodimers during avian skeletal muscle development.

Myosin isoforms contribute to the heterogeneity and adaptability of skeletal muscle fibers. Besides the well-characterized slow and fast muscle myosins, there are those isoforms that appear transiently during the course of muscle development. At a stage of development when two different myosins are coexpressed, the possibility arises for the existence of heterodimers, molecules containing two different heavy chains, or homodimers, molecules with two identical heavy chains. The question of whether neonatal and adult myosin isoforms can associate to form a stable heterodimer was addressed by using stage-specific monoclonal antibodies in conjunction with immunological and electron microscopic techniques. We find that independent of the ratio of adult to neonatal myosin, depending on the age of the animal, the myosin heavy chains form predominantly homodimeric molecules. The small amount of hybrid species present suggests that either the rod portion of the two heavy chain isoforms differs too much in sequence to form a stable alpha-helical coiled coil, or that the biosynthesis of the heavy chains precludes the formation of heterodimeric molecules.

Animals↗
PubMed1991

Assembly of avian skeletal muscle myosins: evidence that homodimers of the heavy chain subunit are the thermodynamically stable form.

Using a double antibody sandwich ELISA we examined the heavy chain isoform composition of myosin molecules isolated from chicken pectoralis major muscle during different stages of development. At 2- and 40-d posthatch, when multiple myosin heavy chain isoforms are being synthesized, we detected no heterodimeric myosins, suggesting that myosins are homodimers of the heavy chain subunit. Chymotryptic rod fragments of embryonic, neonatal, and adult myosins were prepared and equimolar mixtures of embryonic and neonatal rods and neonatal and adult rods were denatured in 8 M guanidine. The guanidine denatured myosin heavy chain fragments were either dialyzed or diluted into renaturation buffer and reformed dimers which were electrophoretically indistinguishable from native rods. Analysis of these renatured rods using double antibody sandwich ELISA showed them to be predominantly homodimers of each of the isoforms. Although hybrids between the different heavy chain fragments were not detected, exchange was possible under these conditions since mixture of biotinylated neonatal rods and fluoresceinated neonatal rods formed a heterodimeric biotinylated-fluoresceinated species upon renaturation. Therefore, we propose that homodimers are the thermodynamically stable form of skeletal muscle myosin isoforms and that there is no need to invoke compartmentalization or other cellular regulatory processes to explain the lack of heavy chain heterodimers in vivo.

Animals↗
PubMed1989

A myosin isoform repressed in hypertrophied ALD muscle of the chicken reappears during regeneration following cold injury.

A library of monoclonal antibodies specific for myosin heavy chain (HC) was used to study myosin expression in regenerating fibers. The response to cold injury of slow skeletal ALD muscle previously induced to eliminate SM1 myosin by weight overload was compared to that of its contralateral control. Native gel electrophoresis combined with immunoblotting demonstrated that slow SM1 myosin HC eliminated from hypertrophic muscle reappeared both at the site of active regeneration and unexpectedly, also distal to the site of injury. The regeneration response of hypertrophied muscles was similar to that of the controls. In addition to SM1 myosin HC, ventricular-like and embryonic/fast isoforms were also expressed in both muscles during the early stages of regeneration and disappeared as the muscle fibers matured. These observations demonstrate that regenerating slow muscle fibers reexpress myosins' characteristic of developing muscle irrespective of the myosin phenotype prior to injury. The reappearance of repressed myosin HC in the hypertrophied ALD muscle is consistent with the presence of newly differentiated myonuclei.

Animals↗
PubMed1989

Distribution of fast myosin heavy chain isoforms in thick filaments of developing chicken pectoral muscle.

Colloidal gold-conjugated monoclonal antibodies were prepared to stage-specific fast myosin heavy chain (MHC) isoforms of developing chicken pectoralis major (PM). Native thick filaments from different stages of development were reacted with these antibodies and examined in the electron microscope to determine their myosin isoform composition. Filaments prepared from 12-d embryo, 10-d chick, and 1-yr chicken muscle specifically reacted with the embryonic (EB165), neonatal (2E9), and adult (AB8) antimyosin gold-conjugated monoclonal antibodies, respectively. The myosin isoform composition was more complex in thick filaments from stages of pectoral muscle where more than one isoform was simultaneously expressed. In 19-d embryo muscle where both embryonic and neonatal isoforms were present, three classes of filaments were found. One class of filaments reacted only with the embryonic antibody, a second class reacted only with the neonatal-specific antibody, and a third class of filaments were decorated by both antibodies. Similar results were obtained with filaments prepared from 44-d chicken PM where the neonatal and adult fast MHCs were expressed. These observations demonstrate that two myosin isoforms can exist in an individual thick filament in vivo. Immunoelectron microscopy was also used to determine the specific distribution of different fast MHC isoforms within individual filaments from different stages of development. The anti-embryonic and anti-adult antibodies uniformly decorated both homogeneous and heterogeneous thick filaments. The neonatal specific antibody uniformly decorated homogeneous filaments; however, it preferentially decorated the center of heterogeneous filaments. These observations suggest that neonatal MHC may play a specific role in fibrillogenesis.

Animals↗
PubMed1988

Expression of fiber type specific proteins during ontogeny of canine temporalis muscle.

The canine masticatory muscles contain a unique adult fiber type composition and different contractile protein isoforms than do adult limb muscles. To determine when these characteristic proteins are expressed during development, samples from canine temporalis (masticatory) and pectineus (limb) muscles were compared between 55 days gestation and 60 days postpartum by histochemical, biochemical, and immunocytochemical analysis. At 55 days gestation and 3 days postpartum, both muscles contained identical histochemical type 2C fibers, native myosin isozymes, and myosin light and heavy chains. By 14 days postpartum, fiber-type expression in these muscles diverged, with resultant formation of type 1 and type 2M fibers in the temporalis muscle and type 1 and 2A fibers in the pectineus muscle. The distinctive myosin isoforms, light chains, and heavy chain of the temporalis muscle were also expressed 2 weeks postpartum. Based on the methods used in this study, we conclude that (1) the temporalis muscle develops from embryonic fibers that initially contain a myosin indistinguishable from embryonic limb muscle fibers, suggesting they have a common precursor, and (2) the myosin light chains and heavy chain unique to the temporalis muscle are initially expressed 2 weeks postpartum.

Animals↗
PubMed1988

Myosin heavy chain expression during development and following denervation of fast fibers in the red strip of the chicken pectoralis.

The myosin heavy chain composition of muscle fibers that comprise the red strip of the pectoralis major was determined at different stages of development and following adult denervation. Using a library of characterized monoclonal antibodies we found that slow fibers of the red strip do not react with antibodies to any of the fast myosin heavy chains of the superficial pectoralis. Immunocytochemical analysis of the fast fibers of the adult red strip revealed that they contain the embryonic fast myosin heavy chain rather than the adult pectoral isoform found throughout the adult white pectoralis. This was confirmed using immunoblot analysis of myosin heavy chain peptide maps. We show that during development of the red strip both neonatal and adult myosin heavy chains appear transiently, but then disappear during maturation. Furthermore, while the fibers of the superficial pectoralis reexpress the neonatal isoform as a result of denervation, the fibers of the red strip reexpress the adult isoform. Our data demonstrate a new developmental program of fast myosin heavy chain expression in the chicken and suggest that the heterogeneity of myosin heavy chain expression in adult fast fibers results from repression of specific isoforms by innervation.

Age Factors↗
PubMed1988

Diversity of fast myosin heavy chain expression during development of gastrocnemius, bicep brachii, and posterior latissimus dorsi muscles in normal and dystrophic chickens.

The expression of fast myosin heavy chain (MHC) isoforms was examined in developing bicep brachii, lateral gastrocnemius, and posterior latissimus dorsi (PLD) muscles of inbred normal White Leghorn chickens (Line 03) and genetically related inbred dystrophic White Leghorn chickens (Line 433). Utilizing a highly characterized monoclonal antibody library we employed ELISA, Western blot, immunocytochemical, and MHC epitope mapping techniques to determine which MHCs were present in the fibers of these muscles at different stages of development. The developmental pattern of MHC expression in the normal bicep brachii was uniform with all fibers initially accumulating embryonic MHC similar to that of the pectoralis muscle. At hatching the neonatal isoform was expressed in all fibers; however, unlike in the pectoralis muscle the embryonic MHC isoform did not disappear. With increasing age the neonatal MHC was repressed leaving the embryonic MHC as the only detectable isoform present in the adult bicep brachii muscle. While initially expressing embryonic MHC in ovo, the post-hatch normal gastrocnemius expressed both embryonic and neonatal MHCs. However, unlike the bicep brachii muscle, this pattern of expression continued in the adult muscle. The adult normal gastrocnemius stained heterogeneously with anti-embryonic and anti-neonatal antibodies indicating that mature fibers could contain either isoform or both. Neither the bicep brachii muscle nor the lateral gastrocnemius muscle reacted with the adult specific antibody at any stage of development. In the developing posterior latissimus dorsi muscle (PLD), embryonic, neonatal, and adult isoforms sequentially appeared; however, expression of the embryonic isoform continued throughout development. In the adult PLD, both embryonic and adult MHCs were expressed, with most fibers expressing both isoforms. In dystrophic neonates and adults virtually all fibers of the bicep brachii, gastrocnemius, and PLD muscles were identical and contained embryonic and neonatal MHCs. These results corroborate previous observations that there are alternative programs of fast MHC expression to that found in the pectoralis muscle of the chicken (M.T. Crow and F.E. Stockdale, 1986, Dev. Biol. 118, 333-342), and that diversification into fibers containing specific MHCs fails to occur in the fast muscle fibers of the dystrophic chicken. These results are consistent with the hypothesis that avian muscular dystrophy is a developmental disorder that is associated with alterations in isoform switching during muscle maturation.

Age Factors↗
PubMed1987

Canine masticatory muscle disorders: a study of 29 cases.

The histopathologic features in temporalis muscle biopsies from 29 dogs with masticatory muscle disorders were characterized and used for their subgrouping: 2 without lesions, 3 with nonspecific changes, 7 with neurogenic atrophy, and 16 with myositis. The immunocytochemical and immunochemical features of the muscle biopsies and sera from those dogs were compared among the histopathologic subgroupings and compared with biopsies and sera from healthy dogs and dogs with polymyositis. Of the 14 biopsies from dogs with masticatory muscle myositis, 12 had immune complexes limited to type 2M fibers, whereas 13 of 16 sera samples had detectable antibodies against type 2M fibers. The immune complex deposition was found only in biopsies of dogs with masticatory muscle myositis, and the antibodies were detected in the sera of only one dog that did not have masticatory muscle myositis. Immunoblot assays revealed that the antibodies were most often directed against a 185 K protein, myosin heavy chain, and a band that appeared to be LC2-M (myosin light chain 2-masticatory).

Animals↗
PubMed1987

Expression of myosin heavy chain isoforms in regenerating myotubes of innervated and denervated chicken pectoral muscle.

Monoclonal antibodies were prepared to stage-specific chicken pectoral muscle myosin heavy chain isoforms. From comparison of serial sections reacted with these antibodies, the myosin heavy chain isoform composition of individual myofibers was determined in denervated pectoral muscle and in regenerating myotubes that developed following cold injury of normal and denervated muscle. It was found that the neonatal myosin heavy chain reappeared in most myofibers following denervation of the pectoral muscle. Regenerating myotubes in both innervated and denervated muscle expressed all of the myosin heavy chain isoforms which have thus far been characterized in developing pectoral muscle. However, the neonatal and adult myosin heavy chains appeared more rapidly in regenerating myotubes compared to myofibers in developing muscle. While the initial expression of these isoforms in the regenerating areas was similar in innervated and denervated muscles, the neonatal myosin heavy chain did not disappear from noninnervated regenerating fibers. These results indicate that innervation is not required for the appearance of fast myosin heavy chain isoforms, but that the nerve plays some role in the repression of the neonatal myosin heavy chain.

Animals↗
PubMed1986

Contractile activity is required for the expression of neonatal myosin heavy chain in embryonic chick pectoral muscle cultures.

The expression of neonatal myosin heavy chain (MHC) was examined in developing embryonic chicken muscle cultures using a monoclonal antibody (2E9) that has been shown to be specific for that isoform (Bandman, E., 1985, Science (Wash. DC), 227: 780-782). After 1 wk in vitro some myotubes could be stained with the antibody, and the number of cells that reacted with 2E9 increased with time in culture. All myotubes always stained with a second monoclonal antibody that reacted with all MHC isoforms (AG19) or with a third monoclonal antibody that reacted with the embryonic but not the neonatal MHC (EB165). Quantitation by ELISA of an extract from 2-wk cultures demonstrated that the neonatal MHC represented between 10 and 15% of the total myosin. The appearance of the neonatal isoform was inhibited by switching young cultures to medium with a higher [K+] which has been shown to block spontaneous contractions of myotubes in culture. Furthermore, if mature cultures that reacted with the neonatal antibody were placed into high [K+] medium, neonatal MHC disappeared from virtually all myotubes within 3 d. The effect of high [K+] medium was reversible. When cultures maintained in high [K+] medium for 2 wk were placed in standard medium, which permitted the resumption of contractile activity, within 24 h cells began to react with the neonatal specific antibody, and by 72 h many myotubes were strongly positive. Since similar results were also obtained by inhibiting spontaneous contractions with tetrodotoxin, we suggest that the development of contractile activity is not only associated with the maturation of myotubes in culture, but may also be the signal that induces the expression of the neonatal MHC.

Animals↗
PubMed1985

Continued expression of neonatal myosin heavy chain in adult dystrophic skeletal muscle.

The expression of myosin heavy chain isoforms was examined in normal and dystrophic chicken muscle with a monoclonal antibody specific for neonatal myosin. Adult dystrophic muscle continued to contain neonatal myosin long after it disappeared from adult normal muscle. A new technique involving western blotting and peptide mapping demonstrated that the immunoreactive myosin in adult dystrophic muscle was identical to that found in neonatal normal muscle. Immunocytochemistry revealed that all fibers in the dystrophic muscle failed to repress neonatal myosin heavy chain. These studies suggest that muscular dystrophy inhibits the myosin gene switching that normally occurs during muscle maturation.

Age Factors↗