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Biomedical subjects

R A Majack

Publications and source records attributed to R A Majack.

31 records · Page 2Linked to original sources
PubMed1987

Induction of thrombospondin messenger RNA levels occurs as an immediate primary response to platelet-derived growth factor.

We have investigated the regulation of mRNA levels for thrombospondin, a platelet-derived growth factor (PDGF)-regulated secreted glycoprotein, in cultures of rat vascular smooth muscle cells (SMC). A thrombospondin cDNA hybridizes to a single 5.5-kilobase SMC message which is greatly induced by serum or PDGF. When quiescent SMC are treated with PDGF, thrombospondin levels are induced rapidly (within 15 min) and in a dose-dependent manner. The induction of thrombospondin message levels parallels PDGF-mediated mitogenesis. PDGF-mediated increases in thrombospondin mRNA can be blocked in the presence of actinomycin D, suggesting that PDGF regulates the thrombospondin message at the level of gene transcription. In the presence of cycloheximide, PDGF "super-induces" the thrombospondin message. The data establish that mRNA levels for thrombospondin are regulated by PDGF in a manner similar to that of c-myc, c-fos, and other growth-regulatory gene products and extend our previous findings that thrombospondin secretion by SMC is dependent on exposure to PDGF. These observations are strongly suggestive of an important role for thrombospondin in the growth response of mesenchymal cells.

Animals↗
PubMed1987

Beta-type transforming growth factor specifies organizational behavior in vascular smooth muscle cell cultures.

In culture, vascular smooth muscle cells (SMC) grow in a "hill-and-valley" (multilayered) pattern of organization. We have studied the growth, behavioral organization, and biosynthetic phenotype of rat aortic SMC exposed to purified platelet-derived growth regulatory molecules. We show that multilayered growth is not a constitutive feature of cultured SMC, and that beta-type transforming growth factor (TGF-beta) is the primary determinant of multilayered growth and the hill-and-valley pattern of organization diagnostic for SMC in culture. TGF-beta inhibited, in a dose-dependent manner, the serum- or platelet-derived growth factor-mediated proliferation of these cells in two-dimensional culture, but only when cells were plated at subconfluent densities. The ability of TGF-beta to inhibit SMC growth was inversely correlated to plating cell density. When SMC were plated at monolayer density (5 X 10(4) cells/cm2) to allow maximal cell-to-cell contact, TGF-beta potentiated cell growth. This differential response of SMC to TGF-beta may contribute to the hill-and-valley pattern of organization. Unlike its effect on other cell types, TGF-beta did not enhance the synthesis of fibronectin or its incorporation into the extracellular matrix. However, the synthesis of a number of other secreted proteins was altered by TGF-beta treatment. SMC treated with TGF-beta for 4 or 8 h secreted markedly enhanced amounts of an Mr 38,000-D protein doublet whose synthesis is known to be increased by heparin (another inhibitor of SMC growth), suggesting metabolic similarities between heparin- and TGF-beta-mediated SMC growth inhibition. The data suggest that TGF-beta may play an important and complex regulatory role in SMC proliferation and organization during development and after vascular injury.

Animals↗
PubMed1986

Control of smooth muscle cell growth by components of the extracellular matrix: autocrine role for thrombospondin.

Addition of platelet-derived growth factor (PDGF) to growth-arrested cultured smooth muscle cells (SMC) induces the synthesis and secretion of thrombospondin (TS), a glycoprotein component of the SMC extracellular matrix in vitro. This induction occurs at PDGF concentrations that are suboptimal for a mitogenic response. In this study we examined the effect of TS on the proliferation of SMC, using a serum-free mitogenesis assay. Addition of either epidermal growth factor (EGF) or purified human platelet TS to quiescent rat vascular SMC did not substantially stimulate mitogenesis; the 30-hr nuclear labeling index increased from a mean of 7% in control cells to 20% for EGF-treated SMC and 17% for cells exposed to TS alone. However, TS and EGF acted synergistically to stimulate DNA synthesis by SMC, increasing the labeling index to 47%. The facilitative effect of TS on EGF-mediated mitogenesis was inhibited by heparin, a known inhibitor of SMC growth and migration that also blocks incorporation of TS into the SMC extracellular matrix. The effect was specific for EGF; TS did not augment the response of cells to insulin or insulin-like growth factor 1. These data establish a functional role for cell-derived TS and provide evidence for the presence of an autocrine, growth-supportive mechanism involving the extracellular matrix. In addition, our experiments support the existence of a novel, heparin-sensitive SMC mitogenic pathway and suggest a mechanism whereby heparin-like molecules may inhibit SMC proliferation.

Animals↗
PubMed1985

Heparin regulates the collagen phenotype of vascular smooth muscle cells: induced synthesis of an Mr 60,000 collagen.

The effect of heparin on the biosynthetic phenotype of rat vascular smooth muscle cells (SMC) was investigated in vitro. Addition of heparin to the culture medium of early passage rat SMC resulted in a marked (3-15-fold) increase of a cell layer-associated Mr 60,000 protein that was sensitive to digestion by purified bacterial collagenase and contained significant amounts of hydroxyproline. Pulse-chase analysis of heparin-treated SMC revealed that the Mr 60,000 collagen was a primary and abundant product of these cells and was not processed extracellularly to a smaller form. The inductive effect of heparin could be mimicked by iota carrageenan or dextran sulfates but not by hyaluronic acid, dermatan sulfate, or chondroitin sulfates. The induction was concentration dependent with a maximal effect observed at a heparin concentration of 10 micrograms/ml. Synthesis of the Mr 60,000 collagen increased 18-24 h after addition of heparin to the cultures. Following induction and subsequent removal of heparin, synthesis of the protein remained maximal for at least 12 h and required 72 h to return to a basal level. These data demonstrate that the biosynthetic phenotype of vascular SMC in vitro can be controlled, at least in part, by heparin and related polyanions and suggest a role for similar molecules endogenous to the vessel wall in the regulation of SMC function.

Animals↗
PubMed1985

Platelet-derived growth factor and heparin-like glycosaminoglycans regulate thrombospondin synthesis and deposition in the matrix by smooth muscle cells.

Platelet-derived growth factor (PDGF), a smooth muscle cell (SMC) mitogen, and heparin-like glycosaminoglycans, known inhibitors of SMC growth and migration, were found to regulate thrombospondin synthesis and matrix deposition by cultured rat aortic SMC. The synthesis and distribution of thrombospondin was examined in growth-arrested SMCs, in PDGF-stimulated SMCs, and in heparin-treated SMCs using metabolic labeling and immunofluorescence techniques. Thrombospondin synthesis in response to purified PDGF occurred within 1 h after addition of growth factor to growth-arrested SMCs, peaked at 2 h, and returned to baseline levels by 5 h. The induction of synthesis of thrombospondin by PDGF was dose dependent, with a maximal effect observed at 2.5 ng/ml. Actinomycin D (2 micrograms/ml) inhibited thrombospondin induction by PDGF, suggesting a requirement for new RNA synthesis. In the presence of heparin and related polyanions, the incorporation of thrombospondin into the SMC extracellular matrix was markedly reduced. This effect was dose dependent with a maximal effect observed at a heparin concentration of 1 microgram/ml. Heparin did not affect the ability of SMCs to synthesize thrombospondin in response to PDGF. We interpret these data to suggest a role for thrombospondin in the SMC proliferative response to PDGF and in the regulation of SMC growth and migration by glycosaminoglycans.

Animals↗
PubMed1984

Inhibition of vascular smooth muscle cell migration by heparin-like glycosaminoglycans.

Previous studies have suggested that heparin-like glycosaminoglycans may be endogenous inhibitors of smooth muscle proliferation in the vessel wall. The purpose of this study was to determine the effects of exogenous glycosaminoglycans on rat vascular (aortic) smooth muscle cell migration following wounding in vitro. Our data indicate that heparin and related molecules (iota carrageenan, dextran sulfate), but not other glycosaminoglycans (hyaluronate, chondroitin, and dermatan sulfates), inhibit smooth muscle cell motility in a cell-specific, dose-dependent, and reversible fashion. The effect of heparin was maximal (60% inhibition) at 10 micrograms/ml; a half-maximal effect was observed at 1 microgram/ml. Heparin did not significantly affect the migration of bovine aortic endothelium or Swiss 3T3 cells. These observations support the concept that heparin-like glycosaminoglycans may be important regulators of vascular smooth muscle cell function.

Animals↗
PubMed1984

Heparin and related glycosaminoglycans modulate the secretory phenotype of vascular smooth muscle cells.

Previous studies have established a role for heparin-like molecules in the regulation of vascular smooth muscle cell growth and migration in vitro. We present data indicating that the secretory phenotype of cultured rat aortic smooth muscle cells can be modulated by exogenous soluble heparin, heparan sulfate, and dermatan sulfate glycosaminoglycans. In the presence of these molecules, smooth muscle cells secrete increased amounts of two noncollagenous proteins (Mr 37,000 and 39,000). This effect can be mimicked by iota carrageenan and dextran sulfate but not by hyaluronic acid, chondroitin-4-sulfate, or chondroitin-6-sulfate. The inductive effect of heparin was dose-dependent and occurred rapidly (within 1 h) with maximal induction (three- to fivefold over controls) occurring after 10-12 h of treatment. The effect was rapidly reversible (within 1 h) and was not altered in the presence of actinomycin D, suggesting regulation at a posttranscriptional level. These data indicate that the biosynthetic expression of specific smooth muscle cell proteins may be determined, at least in part, by components of the smooth muscle cell extracellular matrix.

Animals↗
PubMed1980

An ameloblastoma with myofibroblasts and intracellular septate junctions.

This report represents a review of the ultrastructural features of the simple ameloblastoma. Our ultrastructural data indicate that the stromal component of ameloblastoma is composed of myofibroblasts with associated collagen and basal lamina material. A unique feature of these myofibroblasts is the formation of plaque-like structures on extended cell processes which were identified as intracellular septate junctions. The epithelial component of ameloblastoma was shown to be composed of three distinct cell types which appear to represent a continuum for the ameloblast-like basal cell through an intermediate to an acanthomatous cell. The actual relationship between the three cell types is unclear.

Adult↗
PubMed1980

The bicellular and reflexive membrane junctions of renomedullary interstitial cells: functional implications of reflexive gap junctions.

Renomedullary interstitial cells are loosely organized within the interstitial space surrounding collecting ducts, limbs of Henle, and capillaries of the rat renal medulla. These cells possess long processes, which interact with each other and with cell bodies to form bicellular tight, intermediate, and gap junctions. In addition, both cell bodies and cell processes possess "reflexive" gap and intermediate junctions. Possible functions of renomedullary interstitial cell membrane junctions are discussed. Particular attention is given to a consideration of the functional significance of "reflexive" gap junctions.

Animals↗
PubMed1979

The ultrastructural localization of membrane ATPase in rat thin limbs of the loop of Henle.

The cytochemical distribution of nonspecific membrane ATPase activity in the epithelial membranes of the thin limbs of the loops of Henle of rat nephrons was studied at the ultrastructural level. Membrane ATPase activity was localized in the luminal, lateral, and (to a lesser extent) basal membranes of only the outer medullary segment of the thin descending limbs of long nephrons (Type II epithelium). The reaction product was lacking in the thin limb of short nephrons (Type I epithelium) as well as in the inner medullary descending (Type III epithelium) and ascending (Type IV epithelium) segments of the thin limbs of long nephrons. These data reinforce the concept of thin limb heterogeneity and may indicate a specialized role for the outer medullary segment of thin descending limbs of long nephrons in the concentrating mechanism.

Adenosine Triphosphatases↗
PubMedDate not supplied

Ultrastructural characteristics of endothelial permeability in chronic hypertension.

This study examined characteristics of paracellular and pinocytotic permeability pathways across the middle cerebral artery endothelium of 12- to 16- month-old spontaneously hypertensive rats (SHR). Interendothelial junctions in SHR, like those of age-matched Wistar-Kyoto controls, were impermeable to lanthanum and horseradish peroxidase (HRP) tracers. Freeze-fracture preparations revealed that interendothelial junctions of chronically hypertensive rats are characterized by a twofold increase over controls in the number of tight junctional strands and the mean apical-basal depth. It is believed that this tight junctional hypertrophy may function to increase adhesive forces between neighboring endothelial cells, and may play a role inthe prevention of hypertension-induced paracellular permeability increases. Morphological and tracer studies of pinocytotic pathways indicated that, it the middle cerebral artery, endothelial vesicular transport activity is not increased during chronic hypertension. No evidence was found to indicate the presence of transendothelial permeability channels across control or hypertensive arterial endothelium. Thus, increased transendothelial permeability, commonly observed in acute hypertension, does not appear to occur during chronic hypertension, at least in the middle cerebral artery. Our findings suggest that the arterial endothelium may undergo structural (tight junctional) adaptation in response to prolonged hypertension.

Animals↗