Sertoli cells are the primary site of prodynorphin gene expression in rat testis: regulation of mRNA and secreted peptide levels by cyclic adenosine 3' ,5'-monophosphate analogs in cultured cells.
Prodynorphin is one of three endogenous opioid peptide genes expressed in testis. Through the use of cell fractionation procedures and Northern blot analysis, Sertoli cells were found to be the primary site of prodynorphin mRNA synthesis in rat testis. In situ hybridization of a prodynorphin cRNA probe to fixed adult tissue confirmed this result. Treatment of primary cultures of rat Sertoli cells with a cAMP analog, 8-(4-chlorophenylthio)cAMP, resulted in a transient 5.6-fold increase in steady state prodynorphin mRNA levels relative to those in control cells. This increase was maximal at 48 h of treatment, after which mRNA levels gradually declined. Treatment of Sertoli cells with cAMP analogs resulted in concurrent 2.6-fold decreases in sulfated glycoprotein-2 mRNA levels. Culture medium from Sertoli cells showed a 3.1-fold increase in secreted dynorphin immunoreactivity after treatment with 8-(4-chlorophenylthio)cAMP. Chromatographic analysis indicates that the majority of the immunoreactive dynorphin peptide synthesized in Sertoli cells is present as high mol wt species, with some processing to bioactive peptides.
Collard
M W
MW
Mental Health Research Institute, University of Michigan, Ann Arbor 48109.
Day
R
R
Akil
H
H
Uhler
M D
MD
Douglass
J O
JO
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics; D002845:Chromatography; D051152:Clusterin; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000494:pharmacology; D004745:Enkephalins / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006023:Glycoproteins / Q000235:genetics; D007700:Kinetics; D008297:Male; D018832:Molecular Chaperones; D009693:Nucleic Acid Hybridization; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D012708:Sertoli Cells / Q000378:metabolism*; D013737:Testis / Q000378:metabolism*; D013873:Thionucleotides / Q000494:pharmacology; D014018:Tissue Distribution
D051152:Clusterin; D004745:Enkephalins; D006023:Glycoproteins; D018832:Molecular Chaperones; D011498:Protein Precursors; D012333:RNA, Messenger; D013873:Thionucleotides; C015929:8-((4-chlorophenyl)thio)cyclic-3',5'-AMP; D003994:Bucladesine; C033660:preproenkephalin; D000242:Cyclic AMP; D002786:Cholesterol Side-Chain Cleavage Enzyme