The Phenotypic Complexity of Myogenic Clones

Abbott, Joan; Schiltz, John R.; Dienstman, S.; Holtzer, H.

doi:10.1073/pnas.71.4.1506

Cited by 62 publications

(29 citation statements)

References 23 publications

(18 reference statements)

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“…In standard cultures, only the ceils that fuse to form the multinucleated myotubes can be identified unambiguously as belonging to myogenic lineage. Contrary to the claims of Doering and Fischman (16) or of Buckley and Konigsberg (8), there are no reliable cytological means of distinguishing replicating mononucleated myogenic cells from mononucleated fibrogenic cells or from mononucleated mesenchyme cells (1,14,19). Accordingly, in this report the response of cells to the action of CB or Colcemid will be, for the most part, in terms of mononucleated cells or multinucleated myotubes.…”

Section: Methodsmentioning

confidence: 41%

Response of myogenic and fibrogenic cells to cytochalasin B and to colcemid. I. Light microscope observations.

Croop

Holtzer²

1975

The Journal of Cell Biology

115

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Cytochalasin B (CB) induces a biphasic retraction in some cell types. The rapid response that peaks in 30 min leads to the "dendritic" condition. Replicating myogenic and fibrogenic cells, as well as postmitotic myoblasts and myotubes, participate in this reaction. This is followed by a slower phase that requires 40 h for stabilization and leads to the fully "arborized" state. Only replicating myogenic and fibrogenic cells participate in this reaction. Postmitotic myoblasts and myotubcs do not arborize but round up and float off into the medium. Pretreatment with Colcemid does not block the rapid response to CB, but does block arborization. CB-arborized cells exposed to Colcemid while in the presence of CB develop sufficient tension to pull themselves apart. If CB depolymerizes actin-like filaments, and if such filaments constitute the only contractile system in the cell, then it is difficult to visualize how cells in CB develop such tension.Colcemid induces twisting, birefringent bands in interphase-and metaphasearrested myogenic and fibrogenic cells, and in postmitotic myotubes. Such bands are much more evident when CB-arborized cells are removed from CB and allowed to relax in Colcemid. These birefringent bands assemble in the presence of cycloheximide, and may constitute 20% of the volume of the cell.The conflicting literature on the pleiotropic effects of cytochalasin B (CB) has been reviewed repeatedly (6,10,18,22,32,36,37,44). Reports have stated that the antibiotic (a) interferes with cytokinesis, (b) inhibits the uptake of sugars, (c) blocks amoeboid movement, (d) prevents sorting out of embryonic cells, and (e) disturbs endo-and r Not all of these effects, however, are observed in all cell types (6,13,20,41). There is ample, though by no means unambiguous, literature that many of these effects of CB are due to one reaction, namely the reversible disruption of microfilaments associated with the cell surface (3, 42, 44, 45, 47; see, however, references 6, 18, 24).It has been postulated that these filaments serve as a primitive contractile system, and the observation that heavy meromyosin (HMM) decorates such filaments (28) has been cited by some as evidence supporting this notion (44,45,47).Sanger and Holtzer (40) and Holtzer and Sanger (23), however, have stressed that the addition of CB to muscle cultures did not interfere with either the synthesis or the polymerization of actin into thin filaments, or with its interaction with myosin required for contraction (20,31). CB did, however, block locomotion of presumptive myoblasts and fibroblasts and did block fusion of postmitotic myoblasts into myotubes. With respect to the

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Section: Methodsmentioning

confidence: 41%

Response of myogenic and fibrogenic cells to cytochalasin B and to colcemid. I. Light microscope observations.

Croop

Holtzer²

1975

The Journal of Cell Biology

115

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“…Standard 6-day muscle cultures were prepared by trypsinizing the breast muscles of 10-day chick embryos (13). During the first 2 days these cultures consist largely of presumptive myoblasts, and lesser numbers of myoblasts, fibroblasts, and mesenchyme cells (14). Significant numbers of multinucleated myotubes first appear in these standard cultures on day 3.…”

Section: Methodsmentioning

confidence: 99%

Differences among myosins synthesized in non-myogenic cells, presumptive myoblasts, and myoblasts.

C¹,

Fellini²,

Holtzer³

1975

Proc. Natl. Acad. Sci. U.S.A.

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“…There are several reports that cells in muscle cultures can be infected with either DNA or RNA viruses (1)(2)(3)(4)(5)(6). However, muscle cultures, and even "muscle clones" consist of a heterogeneous population of cells in different compartments of the myogenic and fibrogenic lineages, and it is by no means clear whether these different phenotypes respond to the viruses in the same manner (7). For example, when muscle cultures are treated with oncogenic virus, does the virus transform both replicating myogenic and post-mitotic myogenic cells?…”

mentioning

confidence: 99%

Effect of oncogenic virus on muscle differentiation.

Holtzer¹,

Biehl²,

Yeoh³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

141

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The question whether some events which control differentiation can modulate the course of viral-induced transformation is an important one and the answer to this question may throw some light on the mechanism of transformation as well as differentiation. There are several reports that cells in muscle cultures can be infected with either DNA or RNA viruses (1-6). However, muscle cultures, and even "muscle clones" consist of a heterogeneous population of cells in different compartments of the myogenic and fibrogenic lineages, and it is by no means clear whether these different phenotypes respond to the viruses in the same manner (7). For example, when muscle cultures are treated with oncogenic virus, does the virus transform both replicating myogenic and post-mitotic myogenic cells? Will the post-mitotic nuclei in infected myotubes integrate the virus, transcribe the viral genome, and transform? Will normal, post-mitotic myotube nuclei be induced to reenter the cell cycle and undergo mitosis as claimed by some investigators (3, 6)? Clearly, answers to these questions will contribute to understanding myogenesis, and could provide a useful model for following events responsible for transformation.We have confirmed previous observations on the effect of wild-type Rous sarcoma virus (RSV) on muscle cultures (5). Replicating myogenic cells exposed to RSV fused to form post-mitotic, multinucleated myotubes, but these myotubes invariably vacuolated and degenerated within the next

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The Phenotypic Complexity of Myogenic Clones

Abstract: A single cell isolated from cultured 8-day leg muscle

Cited by 62 publications

References 23 publications

Response of myogenic and fibrogenic cells to cytochalasin B and to colcemid. I. Light microscope observations.

Response of myogenic and fibrogenic cells to cytochalasin B and to colcemid. I. Light microscope observations.

Differences among myosins synthesized in non-myogenic cells, presumptive myoblasts, and myoblasts.

Effect of oncogenic virus on muscle differentiation.

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