Differentiation-controlled synthesis and binding of thrombospondin to granulosa cells.
Thrombospondin (TSP) is a large glycoprotein, synthesized by several matrix-forming cells and incorporated into their extracellular matrix. In several cell types its presence supports cell growth and proliferation. To investigate the role of this protein in cell differentiation, we studied the hormonal effect of TSP production and receptor-mediated binding to primary granulosa cells prepared from diethylstilbestrol-treated immature female rats. These cells can be induced to differentiate by FSH, 8-bromo-cAMP (8-Br-cAMP), or forskolin. Progesterone production is induced during differentiation, and its level of synthesis is an important manifestation of the differentiated phenotype. We find that undifferentiated granulosa cells synthesize and secrete TSP. The protein comprises about 0.5% of the total cell protein, and it is the major protein secreted in culture. Treatment of the cells with FSH or 8-Br-cAMP reduces TSP production dramatically, and forskolin completely inhibits it. In parallel, we observed that the undifferentiated cells bind TSP specifically with a Kd of 1.8 nM, and the number of binding sites per cell is 1.7 x 10(5). This binding can be prevented by excess TSP or an anti-TSP monoclonal antibody (B7-3). This ability to bind TSP is completely lost after induction of differentiation by FSH or 8-Br-cAMP. Our findings show that both the production and binding of TSP to granulosa cells are tightly controlled by normal cell differentiation and indicate that changes in TSP are correlated with the passage of the cell through the stages of maturation, a passage that also involves changes in cell shape and extracellular interactions and in the steroidogenic capacity of these cells.
Dreyfus
M
M
Department of Polymer Research, Weizmann Institute of Science, Rehovot, Israel.
Dardik
R
R
Suh
B S
BS
Amsterdam
A
A
Lahav
J
J
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D001667:Binding, Competitive; D018955:CD36 Antigens; D002454:Cell Differentiation; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D004054:Diethylstilbestrol / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D037102:Lectins / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000096:biosynthesis / Q000378:metabolism*; D051381:Rats; D016024:Receptors, Cytoadhesin / Q000378:metabolism*; D019699:Thrombospondins
D018955:CD36 Antigens; D037102:Lectins; D010980:Platelet Membrane Glycoproteins; D016024:Receptors, Cytoadhesin; D019699:Thrombospondins; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D004054:Diethylstilbestrol