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Identification and Distribution of a Novel Platelet-Derived Growth Factor Receptor ß Variant: Effect of Retinoic Acid and Involvement in Cell Differentiation

Department of Biochemistry and Molecular and Cellular Biology, Georgetown University Medical Center, Washington, D.C. 20057

Address all correspondence and requests for reprints to: Martine Culty, Ph.D., Department of Biochemistry and Molecular and Cellular Biology, Georgetown University Medical Center, Basic Science Building, 3900 Reservoir Road NW, Washington, D.C. 20057. E-mail: cultym{at}georgetown.edu.

We have shown previously that neonatal testicular gonocytes express platelet-derived growth factor receptors (PDGFR) and ß. We report the expression of a novel PDGFRß (V1-PDGFRß) transcript in gonocytes of 3-d-old rat testes. V1-PDGFRß nucleotide sequence spans from intron 6 to exon 23 of the PDGFRß gene, and is predicted to encode a protein lacking part of the extracellular domain. V1-PDGFRß transcripts are expressed preferentially in developing gonads. The embryonic teratocarcinoma F9 cells, in which differentiation is driven by retinoic acid (RA), express V1-PDGFRß, but not wild-type PDGFRß. Green fluorescent protein-tagged V1-PDGFRß localized mainly in cytosol of F9, MA-10, and COS-1 cells. FLAG and green fluorescent protein-tagged V1-PDGFRß displayed tyrosine kinase activities and contain phosphotyrosine residues, suggesting that V1-PDGFRß is a cytosolic tyrosine kinase. Treatment of F9 cells with RA induced V1-PDGFRß gene expression, concomitant with changes in morphology and increased mRNA expression of collagen IV and laminin B1, suggesting that V1-PFGRß is involved in cell differentiation. Similarly, treatment of postnatal d 3 rat gonocytes with RA induced a dose-dependent increase in V1-PDGFRß expression together with an increase in c-kit and Stra8, markers of more differentiated germ cells and a concomitant decrease in GFR1, a marker of spermatogonial stem cells. However, an excess of V1-PDGFRß inhibited RA-mediated collagen IV and laminin B1 expression and altered both RA-dependent and RA-independent morphological changes in F9 cells, while increasing cell survival. These results suggest that the expression of V1-PDGFRß is tightly regulated during differentiation and that it may play an active role in germ cell differentiation.

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