共查询到20条相似文献,搜索用时 15 毫秒
1.
B J Hoerl R E Scott 《Virchows Archiv. B, Cell pathology including molecular pathology》1978,27(4):335-345
The shedding of plasma membrane vesicles has been shown to result from exposure of monolayer cell cultures to formaldehyde and other sulfhydryl blocking agents. Incubation of cells in concentrations of these agents as low as 5 to 10 mM for intervals as brief as fifteen minutes is effective (Scott, 1976). Plasma membrane vesiculation has been shown to be an energy-dependent process that requires Ca++ and physiological temperature. Following plasma membrane vesiculation, cell monolayers appear intact by phase microscopy and show only slight evidence of cell injury by electron microscopy. In view of these observations, the question has been raised whether plasma membrane vesiculation is compatible with continued cell growth and metabolism. The experiments described in this paper were designed to answer these questions. We pulse exposed 3T3 mouse embryo cells to concentrations of formaldehyde, between 2.5 and 250 mM, for intervals 15, 30 or 60 min. Cell momolayers were then washed in a variety of different media in an attempt to reverse the effect of formaldehyde on cells. Cell monolayers were thereafter assayed for the shedding of plasma membrane vesicles and for their ability to transport 2-deoxy-D-glucose. Cells were also replated in serum-containing medium and their ability to grow was assayed over a seven day interval. The results show an inverse relationship between the shedding of plasma membrane vesicles and the ability of the cells to transport nutrients and to grow. We interpret these data to suggest that the process of plasma membrane vesiculation results from a form of cell injury which blocks cellular metabolism and growth. 相似文献
2.
Differences in the cyclic AMP-dependent phosphorylation of plasma membrane proteins of differentiated and undifferentiated L6 myogenic cells 总被引:1,自引:0,他引:1
Differences in the cyclic AMP-dependent plasma membrane phosphorylation system of undifferentiated and differentiated L6 myogenic cells have been detected. Endogenous plasma membrane protein phosphorylation in undifferentiated L6 myoblasts was stimulated more than three fold by 5 x 10(-5) M cyclic AMP, whereas no statistically significant cyclic AMP-dependent phosphorylation of endogenous plasma membrane proteins was observed in differentiated L6 cells. In undifferentiated cells cyclic AMP promoted the phosphorylation of several proteins, the most prominent of which had a molecular weight of 110,000. In differentiated cells cyclic AMP did not selectively promote the phosphorylation of specific plasma membrane proteins. Both differentiated and undifferentiated L6 cells, however, contain a cyclic AMP-dependent protein kinase capable of catalyzing the phosphorylation of exogenous substrates, such as histone f2b. Therefore, the data show that differentiation in L6 cells is associated with a selective change in the activity of a plasma membrane cyclic AMP-dependent protein kinase which employs endogenous membrane proteins as substrate. 相似文献
3.
Cathepsin B localizes to plasma membrane caveolae of differentiating myoblasts and is secreted in an active form at physiological pH 总被引:1,自引:0,他引:1
Jane DT Morvay L Dasilva L Cavallo-Medved D Sloane BF Dufresne MJ 《Biological chemistry》2006,387(2):223-234
Our in vitro studies support a functional link between the induction of cathepsin B gene expression and the catabolic restructuring associated with myotube formation during myogenesis in vivo. We have tested two predictions that are basic to this hypothesis: (1) that active cathepsin B is localized to plasma membrane caveolae of fusing myoblasts; and (2) that active cathepsin B is secreted from fusing myoblasts at physiological pH. During differentiation, L6 rat myoblasts demonstrated a fusion-related increase in activity associated with the 25/26-kDa, fully processed, active form of cathepsin B. Immunocytochemical studies demonstrated a redistribution of lysosomal cathepsin B protein toward the membrane of fusing myoblasts, and a colocalization of cathepsin B with caveolin-3, the muscle-specific structural protein of membrane caveolae. Sucrose density fractionation and Western blot analysis demonstrated that an active form of cathepsin B localizes to caveolar fractions along with caveolin-3, annexin-VII, beta-dystroglycan and dystrophin. Finally, 'real-time' activity assays and Western blot analysis demonstrated that active cathepsin B is secreted from fusing myoblasts at physiological pH. Collectively, these studies support an association of active cathepsin B with plasma membrane caveolae and the secretion of active cathepsin B from differentiating myoblasts during myoblast fusion. 相似文献
4.
P J Bilan Y Mitsumoto F Maher I A Simpson A Klip 《Biochemical and biophysical research communications》1992,186(2):1129-1137
The GLUT3 facilitative glucose transporter protein was found to be expressed in rat L6 muscle cells. It was detected at both the myoblast and myotube stage. GLUT3 protein content per mg of total membrane protein increased significantly during L6 cell differentiation. Subcellular fractionation demonstrated that the GLUT3 protein was predominantly localized in plasma membrane-enriched fractions of either myoblasts or myotubes. Short-term exposure of L6 myotubes to IGF-I or insulin caused a redistribution of GLUT3 protein from an intracellular membrane fraction to the plasma membrane, without affecting total membrane GLUT3 protein content. Long-term exposure of L6 myotubes to IGF-I produced an increase of GLUT3 protein in total membranes and all subcellular membrane fractions, especially the plasma membrane. We propose that the GLUT3 glucose transporter may play an important role in glucose metabolism in developing muscle. 相似文献
5.
A. V. Salova E. A. Leontieva T. P. Mozhenok E. S. Kornilova S. A. Krolenko T. N. Belyaeva 《Cell and Tissue Biology》2011,5(3):255-263
A study of changes in intracellular processes in the course of the differentiation of myoblasts into myotubules is of great
importance for understanding fundamental problems of cell biology. This primarily concerns the change in the spatial organization
of the vacuolar apparatus, which reflects changes in the properties of the membranes, cytoskeleton, and processes of vesicular
transport in the course of differentiation. In the present work, the distribution of acid organelles (lysosomes, late endosomes,
Golgi cisternae) accumulating acridine orange during the process of the formation of myotubules in the L6J1 cell culture is
analyzed. The perinuclear localization of acid cell organelles in myoblasts is shown to be replaced by the diffuse distribution
of these organelles throughout the entire volume of the myotubule cytoplasm. The use of lipophilic dyes RH 414 or di-8-ANEPPS
enables one to compare the process of formation and dynamics of endocytic vesicles in myoblasts and myotubules. For the analysis
of nonspecific endocytosis, we also used the semiconductor nanocrystals, i.e., quantum dots (QDs) conjugated with TAT-peptide,
a cell-penetrating peptide. We have shown that the QD-TAT complexes enter into myoblasts via endocytosis. It has been established
that QD-TAT complexes penetrate myoblasts, but do not penetrate myotubules, even after incubation for 24 h, which can indicate
changes in the properties of the plasma membrane in the process of differentiation of skeletal muscle fibers. 相似文献
6.
Cell adhesion to collagen and decreased myogenic gene expression implicated in the control of myogenesis by transforming growth factor beta 总被引:12,自引:0,他引:12
Transforming growth factor beta 1 (TGF-beta 1) is an inhibitor of skeletal muscle myoblast differentiation. Myoblast differentiation is dependent on the expression of certain myogenic differentiation genes and is affected by cell interaction with the extracellular matrix. We have searched for events in the differentiation process of L6E9 rat myoblasts that may be involved in the inhibitory action of TGF-beta 1. Elevated expression of the myogenic differentiation gene, myogenin, which is thought to play a central role in the differentiation process, occurs 10 h after the shift of L6E9 myoblasts to differentiation medium. Elevation of myogenin mRNA is blocked by TGF-beta 1 added at the time of the shift. This effect is preceded by, and might be related to, a rapid up-regulation of junB mRNA observed in TGF-beta 1-treated L6E9 myoblasts. However, TGF-beta 1 can also block myogenic differentiation in cells transfected with the myogenin gene under the control of a constitutive SV40 viral promoter. The nature of a mechanism that could mediate the inhibitory action of TGF-beta 1 without blocking myogenin mRNA expression is suggested by the observations that (a) TGF-beta 1 upregulates type I collagen expression and deposition in L6E9 myoblasts, (b) a fibrillar type I collagen layer inhibits L6E9 myoblast differentiation, and (c) inhibition of L6E9 myoblast differentiation by a type I collagen layer occurs without a block in myogenin expression. Thus, the data suggest that inhibition of L6E9 myoblast differentiation by TGF-beta 1 may be accomplished by at least two mechanisms acting in concert. One mechanism leads to a block in the expression of myogenin whereas the other mechanism may involve TGF-beta 1-induced changes in cell adhesion that either block the action of myogenic differentiation gene products or prevent the function of other as yet unknown components of the myogenic differentiation pathway. 相似文献
7.
An analog of lysophosphatidylcholine (1-dodecyl-propanediol-3-phosphocholine) which does not impair membrane-bound enzymes was used for the induction of shedding of membrane vesicles from intact calf thymocytes. Without liberation of intracellular enzymes such as lactate dehydrogenase (EC 1.1.1.27) the shedded membranes contained 15--25% of the total activity of the plasma membrane enzymes alkaline phosphatase (EC 3.1.3.1), nucleotide pyrophosphatase (EC 3.1.4.1) and gamma-glutamyl transferase (EC 2.3.2.2). Membrane-free supernatants only exhibited trace activities of these enzymes. Without further purification, the specific enzyme activities in shedded membranes were of the same order of magnitude as in purified plasma membranes prepared after nitrogen cavitation of thymocytes. Small amounts of membrane vesicles which showed a different composition could be removed without detergent. These membranes exhibited a 3-fold lower specific activity of the gamma-glutamyl transferase while that of the alkaline phosphatase and nucleotide pyrophosphatase was similar as in detergent induced membrane vesicles. Distinct differences also were found in the protein pattern. The content of total cholesterol and phospholipid in vesicles shed spontaneously or after detergent treatment was nearly identical, however, significant differences were found in the fatty acid composition of the main phospholipids. The content of polyunsaturated fatty acids (linoleic and arachidonic acid) increased in the order: spontaneously shedded membranes, detergent induced vesicles, conventional purified plasma membranes. These results are discussed in terms of the heterogeneous composition of areas of the thymocyte plasma membrane. 相似文献
8.
D Kim S Chi K H Lee S Rhee Y K Kwon C H Chung H Kwon M S Kang 《The Journal of biological chemistry》1999,274(22):15395-15400
During myogenesis, mononucleated myoblasts form multinucleated myotubes by membrane fusion. Efficiency of this intercellular process can be maximized by a simultaneous progress, with a time window, of other neighboring myoblasts in the differentiation program. This phenomenon has been described as the community effect. It proposes the existence of a molecule that acts as a differentiation-inducing signal to a group of identical cells. Here, we show that neuregulin is a strong candidate for this molecule in myoblast differentiation. The expression of neuregulin increased rapidly but transiently at early stage of differentiation of rat L6 cells. Neuregulin showed a potent differentiation-promoting activity in membrane fusion and expression of myosin heavy chain. The antibodies raised against neuregulin and its cognate receptor ErbB3, which were capable of neutralizing the signal pathway, inhibited myotube formation and expression of myosin heavy chain in both L6 cells and primary rat myoblasts. The progress of differentiation was mostly halted after the expression of myogenin and cell cycle arrest. These results suggest that the activation of an autocrine signaling of neuregulin may provide a basic mechanism for the community effect observed in the differentiation of the embryonic muscle cells. 相似文献
9.
L6 myoblasts spontaneously undergo differentiation and cell fusion into myotubes. These cells express both GLUT1 and GLUT4 glucose transporters, but their expression varies during myogenesis. We now report that the subcellular distribution and the protein processing by glycosylation of both glucose transporter isoforms also change during myogenesis. Crude plasma membrane and light microsome fractions were isolated from either myoblasts or myotubes and characterized by the presence of two functional proteins, the Na+/K(+)-ATPase and the dihydropyridine receptor (DHPR). Immunoreactive alpha 1 subunit of the Na+/K(+)-ATPase was faint in the crude plasma membrane fraction from myoblasts, but abundant in both membrane fractions from myotubes. In contrast, the alpha 1 subunit of the DHPR, which is expressed only in differentiated muscle, was detected in crude plasma membrane from myotubes but not from myoblasts. Therefore, crude plasma membrane fractions from myoblasts and myotubes contain cell surface markers, and the composition of these membranes appears to be developmentally regulated during myogenesis. GLUT1 protein was more abundant in the crude plasma membrane relative to the light microsome fraction prepared from either myoblasts or myotubes. The molecular size in sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the GLUT1 transporters in myotubes was smaller than that in myoblasts (Mr 47,000 and 53,000, respectively). GLUT4 protein (Mr 48,000) was barely detectable in the crude plasma membrane fraction and was almost absent in the light microsome fraction prepared from myoblasts. However, GLUT4 protein was abundant in myotubes and was predominantly located in the light microsome fraction. Treatment with endoglycosidase F reduced the molecular size of the transporters in all fractions to Mr 46,000 for GLUT1 and Mr 47,000 for GLUT4 proteins. In myotubes, acute insulin treatment increased the crude plasma membrane content of GLUT1 marginally and of GLUT4 markedly, with a concomitant decrease in the light microsomal fraction. These results indicate that: (a) the subcellular distribution of glucose transporters is regulated during myogenesis, GLUT4 being preferentially sorted to intracellular membranes; (b) both GLUT1 and GLUT4 transporters are processed by N-linked glycosylation to form the mature transporters in the course of myogenesis; and (c) insulin causes modest recruitment of GLUT1 transporters and marked recruitment of GLUT4 transporters, from light microsomes to plasma membranes in L6 myotubes. 相似文献
10.
The organelles of the exocytic pathway undergo a profound reorganization during the myogenic differentiation. Here, we have
investigated the dynamics of the membrane trafficking at various stages of the differentiation process by using the green
fluorescent protein-tagged, temperature-sensitive vesicular stomatitis virus G protein (tsG-GFP) as a marker. At the restrictive
temperature of 39°C, the tsG-GFP located to the endoplasmic reticulum (ER) at each stage of differentiation. Mobile membrane
containers moving from the ER to the Golgi elements were seen in myoblasts and myotubes upon shifting the temperature to 20°C.
In adult myofibers, in contrast, such containers were not seen although the tsG-GFP rapidly shifted from the ER to the Golgi
elements. The mobility of tsG-GFP in the myofiber ER was restricted, suggesting localization in an ER sub-compartment. Contrasting
with the ER-to-Golgi trafficking, transport from the Golgi elements to the plasma membrane involved mobile transport containers
in all differentiation stages. These findings indicate that ER-to-Golgi trafficking in adult skeletal myofibers does not involve
long-distance moving membrane carriers as occurs in other mammalian cell types. 相似文献
11.
Protein kinase C activity and phorbol ester binding to rat myogenic cells during growth and differentiation 总被引:1,自引:0,他引:1
Phorbol esters have been reported to induce opposite responses in fetal myoblasts and in satellite cells isolated from adult skeletal muscles. We examined the possibility that different levels of protein kinase C (PKC) activity and different phorbol ester binding characteristics account for these responses. For this purpose, the subcellular distributions of PKC were compared in primary cultures of myogenic cells from fetal and adult rat muscles and in the L6 cell line. Cells were used at the proliferative stage or after differentiation into myotubes. Binding of phorbol dibutyrate (PDBu) was assayed. In all three cell types, the levels of PKC specific activity were comparable at the proliferating and the differentiated stages, and partial translocation of PKC activity from the membrane to the cytosolic compartment was observed after differentiation into myotubes. PDBu binding, which had a Kd of 6 to 13 nM in proliferative cells, rose to between 30 and 52 nM in myotubes. Simultaneously, a small increase was observed in the total number of PDBu binding sites. These results suggest that the role of PKC might change with the stage of differentiation. They also imply that the difference described by others between the sensitivity to phorbol esters of fetal myoblasts and satellite cells is not connected with the phorbol ester receptor (i.e., PKC), but might be caused by events subsequent to PKC activation. 相似文献
12.
Role of Ca2+ and Ca2+-activated protease in myoblast fusion 总被引:1,自引:0,他引:1
J E Schollmeyer 《Experimental cell research》1986,162(2):411-422
In this report, we have examined the effects of a calcium chelator, EGTA, and a calcium ionophore, A23187, on fusion of a cloned muscle cell line, L6. Our results confirm that EGTA essentially blocks all myoblast fusion because the lateral alignment of presumptive myoblasts cannot occur in the absence of extracellular calcium. A23187, however, promotes the precocious fusion of myoblasts, apparently by facilitating Ca2+ transport into myoblasts. We have also demonstrated that a Ca2+-activated protease, CAF (mM), appears to relocate in response to the Ca2+ flux, changing from a random, dispersed distribution in proliferative myoblasts to a predominantly peripheral distribution in prefusion myoblasts. Coincident with the mM CAF relocation is an altered distribution of a surface glycoprotein, fibronectin. Extracellular fibronectin is seen in abundance in proliferating myoblasts, but is essentially absent from the surface of fusing myoblasts. We suggest that mM CAF when activated by Ca2+ influx may act to promote the release of fibronectin from the myoblast cell surface, thus providing a mechanism by which the membrane of the fusing myoblast may be rearranged to accommodate fusion. 相似文献
13.
The cultured myoblasts of the rat skeletal muscle cell line L6 proliferate till confluency and then fuse to form myotubes and express a number of muscle-specific proteins. We had shown that this differentiation process is blocked by specific metalloendoprotease inhibitors. We now demonstrate that metabolizing L6 myoblasts and their cell extracts degrade insulin to acid-soluble fragments by a non-lysosomal pathway. About 90% of the insulin-degrading activity residues in the cytoplasm and is due to a 110-kDa enzyme known as the insulin-degrading enzyme. The same metalloendoprotease inhibitors that block the differentiation of L6 myoblasts also inhibit insulin degradation by the metabolizing L6 cells, their cell extracts, and the insulin-degrading enzyme immunoprecipitated from the cytosolic extracts by a monoclonal antibody. These results suggest that the insulin-degrading enzyme is the metalloendoprotease whose activity is required for the initiation of the morphological and biochemical differentiation of L6 myoblasts. 相似文献
14.
《Cell communication & adhesion》2013,20(4):329-343
Fusion of myoblasts to form syncitial muscle cells results from a complex series of sequential events including cell alignment, cell adhesion and cell communication. The aim of the present investigation was to assess whether intercellular communication through gap junctions would be required for subsequent membrane fusion. The presence of the gap junction protein connexin 43 at areas of contact between prefusing rat L6 myoblasts was established by immunofluorescent staining. These myoblasts were dye-coupled, as demonstrated by the use of the scrape-loading/dye transfer technique. L6 myoblast dye coupling was reversibly blocked by heptanol in short term experiments as well as after chronic treatment. After a single addition of 3.5 mM heptanol, gap junctions remained blocked for up to 8 hours, then this inhibitory effect decreased gradually, likely because the alcohol was evaporated. Changing heptanol solutions every 8 hours during the time course of L6 differentiation resulted in a lasting drastic inhibition of myoblast fusion. We further investigated the effect of heptanol and of other uncoupling agents on the differentiation of primary cultures of embryonic chicken myoblasts. These cells are transiently coupled by gap junctions before myoblast fusion and prolonged application of heptanol, octanol and 18-β-glycyrrhetinic acid also inhibited their fusion. The effect of heptanol and octanol was neither due to a cytotoxic effect nor to a modification of cell proliferation. Moreover, heptanol treatment did not alter myoblast alignment and adhesion. Taken together these observations suggest that intercellular communication might be a necessary step for myoblast fusion. 相似文献
15.
Ren Marc M ge Dani le Goudou Christian Giaume Marc Nicolet Fran ois Rieger 《Cell communication & adhesion》1994,2(4):329-343
Fusion of myoblasts to form syncitial muscle cells results from a complex series of sequential events including cell alignment, cell adhesion and cell communication. The aim of the present investigation was to assess whether intercellular communication through gap junctions would be required for subsequent membrane fusion. The presence of the gap junction protein connexin 43 at areas of contact between prefusing rat L6 myoblasts was established by immunofluorescent staining. These myoblasts were dye-coupled, as demonstrated by the use of the scrape-loading/dye transfer technique. L6 myoblast dye coupling was reversibly blocked by heptanol in short term experiments as well as after chronic treatment. After a single addition of 3.5 mM heptanol, gap junctions remained blocked for up to 8 hours, then this inhibitory effect decreased gradually, likely because the alcohol was evaporated. Changing heptanol solutions every 8 hours during the time course of L6 differentiation resulted in a lasting drastic inhibition of myoblast fusion. We further investigated the effect of heptanol and of other uncoupling agents on the differentiation of primary cultures of embryonic chicken myoblasts. These cells are transiently coupled by gap junctions before myoblast fusion and prolonged application of heptanol, octanol and 18-β-glycyrrhetinic acid also inhibited their fusion. The effect of heptanol and octanol was neither due to a cytotoxic effect nor to a modification of cell proliferation. Moreover, heptanol treatment did not alter myoblast alignment and adhesion. Taken together these observations suggest that intercellular communication might be a necessary step for myoblast fusion. 相似文献
16.
Highly purified plasma membranes were obtained from cells of the L6 line at three characteristic stages of myogenesis: Actively proliferating cells; post-mitotic, confluent myoblasts which have already aligned; and fused myotubes. Differential glycosylation of the plasma membrane proteins of these cells was detected by staining polyacrylamide gels of the separated components with three lectins of different specificity: Concanavalin A (conA), wheat germ agglutinin (WGA) and phytohemagglutinin (PHA) Els. Four kinds of developmentally regulated changes could be identified. 1. Those which took place only at confluency (160, 150, 90, 85, 60, 43 and 40 kD for conA binding, 190 kD for WGA binding, 190 and 110 kD for PHA Els binding. 2. Those which took place only at fusion (135, 51.5 and 38 kD for conA, 160 and 150 kD for WGA and 150 kD for PHA Els binding). 3. Those where the phenomena initiated at confluency continue during fusion (66.5 and 32 kD for conA and 120 kD for PHA binding). 4. Those where opposite changes take place at confluency and at fusion (48 kD for conA, 180, 98 and 85 kD for PHA binding). These results suggest that most developmentally regulated changes in glycosylation take place during the first cell-cell contact step of myogenesis. Metabolic labelling experiments showed that, on the contrary, only few alterations in the accumulation of plasma membrane proteins take place prior to the main burst of fusion. 相似文献
17.
Developmental regulation of a cadherin during the differentiation of skeletal myoblasts 总被引:5,自引:0,他引:5
Cadherins are a family of integral membrane glycoproteins which mediate calcium-dependent intercellular adhesion in vertebrate species. Here we present evidence that fusion-competent rat L6 myoblasts express a cadherin (Mr 127 kDa). The levels of this cadherin were found to be developmentally regulated. Maximal levels were expressed prior to fusion. The increase in cadherin levels observed during differentiation was prevented by the differentiation inhibitor, 5-bromo-2'-deoxyuridine. L6 myoblasts grown in the presence of anti-cadherin antibodies exhibited an altered morphology in comparison to control cultures, coupled with decreased myoblast fusion. These data indicate that the developmental regulation of cadherin is part of the program of terminal differentiation of skeletal myoblasts, and that cadherins are involved in the process of myoblast fusion. 相似文献
18.
O Mesmer M O Cheung T D'Amore T C Lo 《Biochemical and biophysical research communications》1986,141(1):124-130
Hexose transport in plasma membrane vesicles prepared from L6 rat myoblasts was shown to be stereospecific, activated by glucose starvation and occurred by both high and low affinity systems. Transport by the high affinity system was shown to occur by an active transport process. Furthermore, the high affinity system was shown to be defective in vesicles prepared from F72 cells (hexose transport mutant). These results indicate that the high affinity hexose transport system is retained in the plasma membrane vesicles. Thus plasma membrane vesicles could be of value in further characterization of the L6 high affinity hexose transport system, without interference from the various metabolic events occurring in whole cells. 相似文献
19.
Isolation and preliminary characterisation of DNA-protein complexes from the mitochondria of Saccharomyces cerevisiae 总被引:1,自引:0,他引:1
Four independent rat L6 myoblast cell lines have been selected in a single step for resistance to the cytotoxic effects of the lectin concanavalin A (conA). In contrast to parental wild-type myoblast lines, all of the variant clones are unable to undergo normal cellular differentiation to form multinucleated myotubes or biochemical differentiation to produce an increase in the specific activity of the muscle-specific enzyme, creatine phosphokinase (CPK). The correlation between lectin resistance and loss of fusion potential is very tight; clonal variation studies show that there is less than a 2.8×10?8 chance that the two are not directly related. Membrane preparations from the conA-resistant myoblast lines incorporate significantly less GDP-[14C]mannose into the lipid intermediates of protein glycosylation than preparations from parental wild-type cells. Also, conversion of mannose label to fucose occurs in myoblasts and this pathway is more active in conA-resistant cells than wild-type cells. Reduced binding of labelled conA to the cell surfaces of variant myoblasts was observed which may result from alterations to membrane glycoprotein receptors. These studies suggest that mannosylated glycoproteins of the cell surface play a role in the development of the myotubes from myoblasts. Lectin-resistant myoblasts should be useful model systems for investigating what appears to be a pleiotropic mutation affecting the myogenesis process through membrane modifications. 相似文献
20.
L T Furcht G Wendelschafer-Crabb 《Differentiation; research in biological diversity》1978,12(1):39-45
These studies have examined the distribution of plasma membrane intramembranous particles (PMP) visualized by freeze fracture and concanavalin A receptors seen by ultrastructural cytochemistry of differentiated and undifferentiated L6 myoblasts. Undifferentiated mononucleated cells have a clustered distribution of PMP on the majority of the fracture faces. Associated with cell differentiation and cell fusion a more uniform distribution of PMP is observed. Changes also occur with myoblast differentiation in the topography and dynamics of receptors bound to concanavalin A. If undifferentiated or differentiated cells are fixed with glutaraldehyde and then reacted with con-A a uniform distribution of con-A is seen on the cell surfaces. In contrast to this if unfixed live cells are reacted at 37 degrees C with con-A a profound redistribution occurs on differentiated cells (greater than 99% showing redistribution) while receptors remain in a uniform array on undifferentiated cells (approximately 95% uniform distribution). In addition to the membrane binding, con-A is observed to bind to an extracellular filamentous matrix seen in high density undifferentiated cultures which then appears to be degraded with differentiation and myoblast fusion. These studies show that a number of membrane changes, both structural and dynamic occur with myoblast differentiation. 相似文献