Biomedical subjects

M M Thompson

Publications and source records attributed to M M Thompson.

179 records · Page 10Linked to original sources

PubMed1989

Differential effect of platelet-derived growth factor- versus serum-induced growth on smooth muscle alpha-actin and nonmuscle beta-actin mRNA expression in cultured rat aortic smooth muscle cells.

Previous studies have demonstrated that rat aortic smooth muscle cells (SMC) show marked changes in smooth muscle (SM) alpha-actin content and fractional synthesis as a function of cell density and growth (Owens, G. K., Loeb, A., Gordon, D., and Thompson, M. M. (1986) J. Cell Biol. 102, 343-352; Blank, R., Thompson, M. M., and Owens, G. K. (1988) J. Cell Biol. 107, 299-306). Results of this study show that, although there is a 6-fold increase in SM alpha-actin content in postconfluent density arrested cultures as compared to proliferating subconfluent cultures, SM alpha-actin mRNA levels are not different between these cells. This suggests that the SM alpha-actin gene is constitutively active under both of these conditions and that accumulation of SM alpha-actin in postconfluent cells is due to translational and/or post-translational controls. The relationship between growth and cytodifferentiation was further explored by examining the effects of platelet-derived growth factor (PDGF)- or serum-induced growth on actin expression in postconfluent, quiescent cultures maintained in a defined serum-free media. Although both factors have been shown to stimulate proliferation and decrease fractional SM alpha-actin synthesis (Blank et al., 1988), their effects on actin mRNA levels were quite different. PDGF was found to induce a dramatic drop in SM alpha-actin steady state mRNA level but had no effect on nonmuscle beta-actin mRNA level. In contrast, serum stimulation was shown to increase nonmuscle beta-actin mRNA level, whereas SM alpha-actin mRNA level remained constant. Taken together these results indicate that PDGF is a specific and potent repressor of SM alpha-actin expression in vascular SMC and implicate a possible developmental role for PDGF in control of SMC differentiation. In addition, the observation that the level of SM alpha-actin mRNA is unaltered in serum-stimulated cells indicates that an absolute decrease in SM alpha-actin mRNA is not obligatory for cell cycle entrance.

Actins↗

PubMed1989

Heme inhibits cartilage metabolism and growth in vitro.

The present study was designed to investigate the potential role of heme as a direct inhibitor of cartilage metabolism and growth. We used the embryonic chick pelvic rudiment bioassays and the hypophysectomized rat cartilage sulfation bioassay, both sensitive to growth factors and growth inhibitors. In the chick bioassay pelvic rudiment growth was inhibited when heme was added to the culture medium at 0.1 mmol/L (after five days in culture, cartilage weight 224% +/- 11% of initial with normal serum, but 141% +/- 3% of initial with serum plus heme 0.25 mmol/L); the heme-induced growth inhibition was promptly reversible when cartilages were placed in heme-free medium. This was due, at least in part, to heme-induced inhibition of (1) [35S] sulfate incorporation into proteoglycans by as little as 0.05 mmol/L heme (66% +/- 5% of assay buffer); (2) [14C] leucine incorporation into proteins by greater than or equal to 0.05 mmol/L heme (85% +/- 6% of assay buffer); and (3) [3H] uridine incorporation by greater than or equal to 0.10 mmol/L heme (50% +/- 4% of assay buffer). In the rat cartilage sulfation bioassay (the literature "standard" bioassay for skeletal growth studies) a dose-dependent inhibition of [35S] sulfate incorporation occurred with greater than or equal to 0.01 mmol/L added heme (82% +/- 3% of assay buffer). Heme caused a dose-dependent and reversible inhibition of cartilage metabolism and growth, and it may have a novel role in the pathophysiology of growth retardation associated with some chronic disease.

Animals↗

PubMed1989

Hemoglobin as a direct inhibitor of cartilage growth in vitro.

Growth retardation is a feature of several diseases associated with chronic hemolysis (i.e., uremia and the hemoglobinopathies). Although the growth failure is undoubtedly multifactorial, circulating hemoglobin (Hb) may inhibit cartilage growth directly. We tested this hypothesis using the hypophysectomized rat costal cartilage sulfation bioassay and the embryonic chicken pelvic rudiment bioassay, both very sensitive to growth factors and growth inhibitors. In the rat bioassay, Hb produced a dose-dependent inhibition of both basal and normal rat serum (NRS)-stimulated 35SO4 uptake. In the chick bioassay, NRS stimulated cartilage growth as expected, but Hb severely inhibited both basal and NRS-stimulated growth. However, after the cartilages were preincubated with Hb for 2 days, subsequent exposure to NRS allowed them to resume growth at the same rate as cartilage exposed to NRS for the entire 5 days. The growth inhibition could be accounted for by the heme contained in Hb. We conclude that Hb produces a dose-dependent and reversible inhibition of cartilage growth and may contribute to the growth retardation associated with chronic hemolytic conditions.

Animals↗

PubMed1989

Heart disease risk factors, diabetes, and prostatic cancer in an adult community.

The authors studied the association between heart disease and prostatic cancer in Rancho Bernardo, California, from August 1972 to June 1987. During a 14-year follow-up of 1,776 men aged 50-84 years, 100 cases of prostatic cancer were identified, of which 54 were incident. Systolic blood pressure, cigarette smoking, and plasma cholesterol levels in cases did not differ from those in noncases. After adjustment for age and on multivariate analysis, incident cases of prostatic cancer had a nonsignificantly higher frequency of reported heart disease at baseline. History of diabetes was inversely associated with total prostatic cancer (age-adjusted estimated relative risk = 0.2, 90% confidence interval: 0.0-0.8; multivariate-adjusted relative risk = 0.3, 90% confidence interval: 0.1-1.0). The association between heart disease and prostatic cancer is compatible with diagnostic detection bias. The inverse association between diabetes and prostatic cancer is compatible with a cancer-promoting role for endogenous testosterone, the level of which is lower in diabetics, or a risk-reducing effect of antidiabetic diet or drug therapy.

Age Factors↗

PubMed1988

Cell cycle versus density dependence of smooth muscle alpha actin expression in cultured rat aortic smooth muscle cells.

Cultured smooth muscle cells (SMC) undergo induction of smooth muscle (SM) alpha actin at confluency. Since confluent cells exhibit contact inhibition of growth, this finding suggests that induction of SM alpha actin may be associated with cell cycle withdrawal. This issue was further examined in the present study using fluorescence-activated cell sorting of SMC undergoing induction at confluency and by examination of the effects of FBS and platelet-derived growth factor (PDGF) on SM alpha actin expression in postconfluent SMC cultures that had already undergone induction. Cell sorting was based on DNA content or differential incorporation of bromodeoxyuridine (Budr). The fractional synthesis of SM alpha actin in confluent cells was increased two- to threefold compared with subconfluent log phase cells, but no differences were observed between confluent cycling (Budr+) and noncycling (Budr-) cells. In cultures not exposed to Budr, confluent cycling S + G2 cells exhibited similar induction. These data indicate that cell cycle withdrawal is not a prerequisite for the induction of SM alpha actin synthesis in SMC at confluency. Growth stimulation of postconfluent cultures with either FBS or PDGF resulted in marked repression of SM alpha actin synthesis but the level of repression was not directly related to entry into S phase in that PDGF was a more potent repressor of SM alpha actin synthesis than was FBS despite a lesser mitogenic effect. This differential effect of FBS versus PDGF did not appear to be due to transforming growth factor-beta present in FBS since addition of transforming growth factor-beta had no effect on PDGF-induced repression. Likewise, FBS (0.1-10.0%) failed to inhibit PDGF-induced repression. Taken together these data demonstrate that factors other than replicative frequency govern differentiation of cultured SMC and suggest that an important function of potent growth factors such as PDGF may be the repression of muscle-specific characteristics.

Actins↗

PubMed1986

Developmental changes in isoactin expression in rat aortic smooth muscle cells in vivo. Relationship between growth and cytodifferentiation.

There is an inverse relationship between cellular proliferation and smooth muscle alpha-isoactin expression in cultured vascular smooth muscle cells (SMCs) (Owens, G.K., Loeb, A., Gordon, D., and Thompson, M.M. (1986) J. Cell Biol. 102, 343-352). In the present studies, changes in isoactin expression were studied during developmental growth of rat aortic SMCs (ages 1-180 days) to better understand interrelationships between growth and cytodifferentiation in these cells in vivo. Actin expression (i.e. content and synthesis) was evaluated by one- and two-dimensional gel electrophoresis and using isoactin-specific antibodies. The major actin present in cells from newborn rats was nonmuscle beta-actin (56% of total actin), whereas cells from adult animals contained principally smooth muscle alpha-actin (Sm-alpha-actin) (76% of total actin). Increases in Sm-alpha-actin content with increasing age were due, in part, to an increase in Sm-alpha-actin synthesis. However, in SMCs from 90- and 180-day-old rats, the fractional content of Sm-alpha-actin exceeded its fractional synthesis at a time when total Sm-alpha-actin content was increasing. This suggests that Sm-alpha-actin turns over more slowly in mature animals. Decreases in the frequency of SMCs undergoing DNA synthesis with age could not account for increases in Sm-alpha-actin expression with age. However, combined immunocytological and [3H]thymidine autoradiographic studies demonstrated that nearly 50% of the medial derived cells from newborn rat aortas did not show detectable staining with a monoclonal antibody to smooth muscle-specific isoactins, and the replicative frequency was much higher in these cells than in cells that contained Sm-alpha-isoactins. Taken together, the results of the present studies and previous studies in cultured SMCs support the hypothesis that cessation of proliferation during development is associated with the induction of Sm-alpha-actin expression, but that factors other than cellular growth state play an important role in determining the level of Sm-alpha-actin expression in fully differentiated SMCs.

Actins↗

PubMed1986

Expression of smooth muscle-specific alpha-isoactin in cultured vascular smooth muscle cells: relationship between growth and cytodifferentiation.

The relationship between growth and cytodifferentiation was studied in cultured rat aortic smooth muscle cells (SMCs) using expression of the smooth muscle (SM)-specific isoactins (Vanderkerckhove, J., and K. Weber, 1979, Differentiation, 14:123-133) as a marker for differentiation in these cells. Isoactin expression was evaluated by: (a) measurements of fractional isoactin content and synthesis ([35S]methionine incorporation) by densitometric evaluation of two-dimensional isoelectric focusing sodium dodecyl sulfate gels, and (b) immunocytological examination using SM-specific isoactin antibodies. Results showed the following: (a) Loss of alpha-SM isoactin was not a prerequisite for initiation of cellular proliferation in primary cultures of rat aortic SMCs. (b) alpha-SM isoactin synthesis and content were low in subconfluent log phase growth cells but increased nearly threefold in density-arrested postconfluent cells. Conversely, beta-nonmuscle actin synthesis and content were higher in rapidly dividing subconfluent cultures than in quiescent postconfluent cultures. These changes were observed in primary and subpassaged cultures. (c) alpha-SM actin synthesis was increased by growth arrest of sparse cultures in serum-free medium (SFM; Libby, P., and K. V. O'Brien, 1983, J. Cell. Physiol., 115:217-223) but reached levels equivalent to density-arrested cells only after extended periods in SFM (i.e., greater than 5 d). (d) SFM did not further augment alpha-SM actin synthesis in postconfluent SMC cultures. (e) Serum stimulation of cells that had been growth-arrested in SFM resulted in a dramatic decrease in alpha-SM actin synthesis that preceded the onset of cellular proliferation. These findings demonstrate that cultured vascular SMCs undergo differential expression of isoactins in relation to their growth state and indicate that growth arrest promotes cytodifferentiation in these cells.

Actins↗

PubMed1986

Two different heavy chains are found in smooth muscle myosin.

Two putative myosin heavy chains designated SM1 and SM2 were detected on a 3.5% polyacrylamide-sodium dodecyl sulfate gel electrophoresis system loaded with homogenates of several mammalian smooth muscles. The two polypeptides were present in nearly equal amounts in all smooth muscle tissues tested and in myosin purified from swine carotid media and stomach. Both proteins were equally stained by smooth muscle-specific myosin antibodies. The smaller of the polypeptides had a mobility nearly identical to that of the single heavy chain observed in purified fast-twitch skeletal myosin. Electrophoresis of pyrophosphate extracts from swine carotid media, swine stomach, rabbit thoracic aorta, and guinea pig taenia coli on nondenaturing pyrophosphate gels revealed a single protein band. When subsequently electrophoresed on a sodium dodecyl sulfate gel, the native bands from swine tissue extracts revealed the two putative heavy chains in nearly equal amounts, as well as a large amount of a higher molecular weight peptide whose properties reflect those of filamen. Sodium dodecyl sulfate gel analysis of the myosin band from pyrophosphate gels of purified swine stomach myosin showed exclusively the two heavy chains in a nearly 1:1 ratio. Smooth muscle myosin migrates homogeneously on pyrophosphate gels, and the virtual equality of the two heavy chains may reflect the presence of large amounts of a myosin isoenzyme, which is a heavy-chain heterodimer.

Animals↗

PubMed1985

Host range of mink cell focus-inducing viruses.

The species host range of the recombinant, mink cell focus-inducing (MCF) class of murine retroviruses was determined in vitro and compared to the host range properties of xenotropic and amphotropic murine viruses. In contrast to xenotropic and amphotropic viruses, MCF viruses were restricted in the number of mammalian species they would infect. Cell lines from mouse, rat, mink, ferret, and cat were susceptible to MCF infection and certain virus isolates could infect rabbit cells, but cells from Chinese hamster, buffalo, bat, dog, monkey, and human were resistant to infection by most MCF viruses. The resistance of some of the latter cells was abrogated by phenotypic mixing with xenotropic virus, which demonstrated that MCF species host range was mediated by virus envelope-cell surface interaction. The host range uniformity of the various MCF isolates and the unique species distribution of sensitivity are consistent with the conclusion from other evidence that the MCF viruses comprise a class distinct from xenotropic and amphotropic viruses.

Animals↗

PubMed1976