Expression of glypican-1, syndecan-1 and syndecan-4 mRNAs protein kinase C-regulated in rat immature Sertoli cells by semi-quantitative RT-PCR analysis.
In seminiferous tubules, Sertoli cells provide structural and nutritional support for the developing germinal cells. Cell to cell signalization and cell adhesion require proteoglycans expressed at the cell membrane. A preliminary biochemical and structural approach indicated that cell surface proteoglycans are mostly heparan sulfate (HSPG) in immature rat Sertoli cells. The present study focused on the qualitative and quantitative expression of three membrane HSPG, syndecan-1, syndecan-4 and glypican-1 in Sertoli cells of 20-day-old rat. A semi-quantitative multiplex RT-PCR strategy was developed to appreciate the effect of PKC activation on the mRNA expression of the three HSPG. Our data show that the syndecan-1 and glypican-1 mRNA expression is increased by the phorbol myristate acetate (PMA) suggesting a regulation of their expression by the phosphatidyl inositol pathway, as previously hypothesized (Fagen et al., Biochim. Biophys. Acta, 1472 (1999) 250-261). In addition, a physiological effector of the PKC as ATP gave similar effects. Thus, this over-expression could be related with paracrine factors secreted by germ cells.
Brucato
S
S
Laboratoire de Biochimie IRBA, UPRES A 2608 CNRS, Université de Caen, Esplanade de la Paix, F-14032, Caen, France.
Harduin-Lepers
A
A
Godard
F
F
Bocquet
J
J
Villers
C
C
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D017931:DNA Primers; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D019812:Heparan Sulfate Proteoglycans / Q000235:genetics*; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011509:Proteoglycans / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012708:Sertoli Cells / Q000378:metabolism*; D053668:Syndecan-1; D053671:Syndecan-4; D053667:Syndecans; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D017931:DNA Primers; D019812:Heparan Sulfate Proteoglycans; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D012333:RNA, Messenger; C508243:Sdc1 protein, rat; C508307:Sdc4 protein, rat; D053668:Syndecan-1; D053671:Syndecan-4; D053667:Syndecans; D000255:Adenosine Triphosphate; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate