PubMed Health⌕ Search

Biomedical subjects

H Holtzer

Publications and source records attributed to H Holtzer.

At least 109 records · Page 6Linked to original sources
PubMed1976

Differences among sulfated proteoglycans synthesized in nonchondrogenic cells, presumptive chondroblasts, and chondroblasts.

The sulfated proteoglycans synthesized by definitive chondroblasts in cultured 10-day chick vertebral or epiphyseal cartilages were characterized by their sedimentation profile in a sucrose gradient and their susceptibility to chondroitinase ABC (EC 4.2.2.4; chondroitin ABC lyase). These sulfated proteoglycans were indistinguishable from those synthesized by definitive chondroblasts that emerge from older cultures of somites plus notochord or in older cultures of limb buds. The sulfated proteoglycans of these definitive chondroblasts are readily distinguished from those synthesized by their mother cells, the presumptive chondroblasts, or those synthesized by dedifferentiated or bromodeoxyuridine-suppressed chondroblasts. However, the sulfated proteoglycans synthesized by presumptive chondroblasts or by dedifferentiated or bromodeoxyuridine-suppressed chondroblasts cannot be dintinguished by these techniques from those synthesized by (i) blastodisc cells, (ii) fibroblasts, (iii) spinal cord cells, or (iv) skeletal, cardiac, or smooth muscle cells. Addition of glycosaminoglycans or collagen to the medium did not induce somite or limb presumptive chondroblasts to synthesize the chondroblast-unique sulfated proteoglycans. Cells moving from the presumptive chondroblast compartment into the chondroblast compartment acquire not only the option to initiate the synthesis of chondroblast-unique collagen chains, but also the capacity to synthesize chondroblast-unique sulfated proteoglycans.

Animals↗
PubMed1975

Thick and thin filaments in postmitotic, mononucleated myoblasts.

Addition of cytochalasin B to primary muscle cultures allows the physical separation of postmitotic myogenic cells from replicating presumptive myoblasts and replicating fibroblasts. Mononucleated, postmitotic myoblasts proceed without fusion to synthesize myosin and actin and to assemble these proteins into thick and thin filaments. Although sarcomeres oriented in tandem are not evident and A, H, and I bands are atypical in these mononucleated myoblasts, the irregularly scattered clusters of myofilaments are assembled into remarkably normal interdigitating arrays. These scattered clusters of stacked thick and thin filaments permit the cell to contact spontaneously in the presence of cytochalasin B.

Actins↗
PubMed1975

Effect of oncogenic virus on muscle differentiation.

Chick muscle cultures infected with wild-type Rous sarcoma virus form myotubes, but these myotubes vacuolate and by day 6 most have degenerated, leaving only large numbers of transformed mononucleated, replicating cells. Muscle cultures infected with a temperature-sensitive mutant (TS) at permissive temperatures behave as cells infected with wild-type Rous sarcoma virus. TS-infected cells reared for 8 days at nonpermissive temperature form contracting myotubes, plus large numbers of fibroblastic cells. If these cultures are lowered to permissive temperature, within 72 hr the myotubes vacuolate and degenerate, whereas the mononucleated cells transform. If replicating TS-transformed cells after 8 days at permissive temperature are shifted to nonpermissive temperature, within 72 hr many cells fuse and form contracting, post-mitotic myotubes. Creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2) levels parallel the formation and degeneration of myotubes during these temperature shifts. If the viral transforming gene is expressed in the post-mitotic myotubes it is lethal, whereas it is not lethal if expressed in replicating percursor myogenic cells. The viral gene expression at permissive temperature blocks further myogenesis depending on the position of the cells in the myogenic program. The virus does not cancel the replicating, transformed myogenic cells' commitment to, or position in, the myogenic lineage. When the transforming action of the virus is suppressed, the normal myogenic program resumes.

Animals↗
PubMed1975

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

Myosins synthesized in non-myogenic cells and replicating presumptive myoblasts differ from those synthesized in postmitotic mononucleated myoblasts and myotubes. Myoblasts and myotubes synthesize the definitive light chains, MLC1 and MLC2. These light chains display different molecular weights in sodium dodecyl sulfate-polyacrylamide gels from the fibroblast light chains FLC1 and FLC2 synthesized in non-myogenic cells and presumptive myoblasts. There are immunological differences between the myosin heavy chains synthesized in myoblasts and myotubes and those synthesized in non-myogenic cells and presumptive myoblasts. Fluorescein-labeled antibodies against skeletal light meromyosin are bound only along the lateral edges of emerging and definitive A-bands. This antibody to light meromyosin is not bound to the outside of, or the microfilaments subtending, the plasma membrane in non-myogenic cells or in myoblasts or in myotubes. These findings suggest that: (1) non-myogenic cells and replicating presumptive myoblasts synthesize similar myosin heavy and light chains; (2) replicating presumptive myoblasts synthesize a different set of myosins from those synthesized by their postmitotic daughters, the myoblasts; (3) the myosins associated with the plasma membranes of non-myogenic and myogenic cells are products of structural genes distinct from those coding for the myosins for skeletal myofibrils.

Animals↗
PubMed1975

Effects of cytochaslasin B and colcemide on myogenic cultures.

Muscle cultures treated with cytochalasin B yield mono- and oligonucleated cells of two kinds: (i) arborized, replicating precursor myogenic cells and fibroblasts; and (ii) round, post-mitotic, terminally differentiating myoblasts and myotubes. The arborized cells do not bind fluorescein-labeled antibody against myosin, do not contract rhythmically, and do not display hexagonally stacked thick and thin filaments. The round, mono-nucleated myoblasts and round, oligonucleated myotubes bind the fluorescein-labeled antibody against myosin, contract rhythmically, and display clusters of hexagonally-stacked thick and thin filaments. When cytochalasin B is removed and replaced by colcemide, the arborized cells, but not the post-mitotic muscle cells, acquire a radial symmetry and are induced to assemble massive, meandering cables that may occupy over 25% of the cell volume. These tortuous calbes are positively birefringent and consist exclusively of enormous numbers of 100-A, intermediate-sized filaments.

Animals↗
PubMed1975

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

Cytochalasin B (CB) induces a biphasic retraction is 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 "absorized" state. Only replicating myogenic and fibrogenic cells participate in this reaction. Postmitotic myoblasts and myotubes 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 metaphase-arrested myogenic and fibrogenic cells, and in postmitotic myotubes. Such bands are more evident when CB-arborized cells are removed from CB and allowed to relax in Colcemid. These birefringent bands assemble in the prescence of cycloheximide, and may constitute 20% of the volume of the cell.

Animals↗
PubMed1975

Synthesis of myosin heavy and light chains in muscle cultures.

The weight ratio of myosin/actin, the myosin heavy chain content as the percentage of total protein (wt/wt), and the kinds of myosin light chains were determined in (a) standard muscle cultures, (b) pure myotube cultures, and (c) fibroblast cultures. Cells for these cultures were obtained from the breast of 11-day chick embryos. Standard cultures contain, in addition to myotubes, large numbers of replicating mononucleated cells. By killing these replicating cells with cytosine arabinoside, pure myotube cultures were obtained. The myosin/actin ratio (wt/wt) for pure myotube, standard muscle, and fibroblast cultures average 3.1, 1.9, and 1.1 respectively. By day 7, myosin in myotube cultures represents a minimum of 7% of the total protein, but about 3% in standard cultures and less than 1.5% in fibroblasts cultures. Myosin from standard cultures contains light chain LC1, LC2, and LC3, with a relative stoichiometry of the molarity of 1.0:1.9:0.5 and mol wt of 25,000, 18,000 and 16,000 daltons, identical to those in adult fast muscle. Myosin from pure myotubes exhibits light chains LC1 and LC2, with a molar ratio of 1.5:1.6. Myosin from fibroblast cultures possesses two light chains with a stoichiometry of 1.8:1.8 and mol wt of 20,000 and 16,000 daltons. Clearly, the faster migrating light chain, LC3, found in standard cultures is synthesized not by the myotubes but ty the mononucleated cells. In myotubes, both the assembly of the sarcomeres and the interaction between thick and thin filaments required for spontaneous contraction occur in the absence of light chain LC3. One set of structural genes for the myosin light and heavy chains appears to be active in mononucleated cells, whereas another set appears to be active in multinucleated myotubes.

Adenosine Triphosphatases↗
PubMed1974

The phenotypic complexity of myogenic clones.

A single cell isolated from cultured 8-day leg muscle may, when subcultured, yield a myogenic clone. A myogenic clone consists of myotubes and mononucleated cells. When such a myogenic clone is subcultured, large numbers of mononucleated cells are recovered. These mononucleated cells histologically and biochemically are indistinguishable from authentic fibroblasts cultured under the same conditions: they synthesize (alpha1)(2)alpha2 chains of collagen, large amounts of hyaluronic acid, and modest amounts of chondroitin sulfate. These mononucleated cells, however, will not chondrify when grown under culture conditions known to permit presumptive chondroblasts to differentiate terminally. These findings demonstrate that there is a population of single cells in 8-day muscle that is neither a myoblast nor a fibroblast, but is the common progenitor for cells in the myogenic and fibrogenic lineages: this progenitor, however, is beyond the point of readily yielding chondrogenic cells. These findings are discussed in terms of the limited number of phenotypic options open to differentiating cells in each of the successive compartments of their respective lineages.

Animals↗