Synaptosome-associated protein of 25 kilodaltons in oocytes and steroid-producing cells of rat and human ovary: molecular analysis and regulation by gonadotropins.
The synaptosome-associated protein of 25 kDa (SNAP-25) is crucially involved in exocytosis in neurons. The aim of this study was to investigate whether it is present in the ovary. We found SNAP-25 to be expressed in nonneuronal cells of the rat and human ovary, namely in all oocytes and in steroidogenic cells, including granulosa cells (GC) of large antral follicles and luteal cells. Both isoforms, SNAP-25a and b, were found in the ovary. Oocytes obtained by laser capture microdissection were shown to express SNAP-25b, whereas SNAP-25a was found in rat GC and human luteinized GC. Immunohistochemical observations of strong SNAP-25 staining in GC of large growing antral follicles compared with absent or weak staining in small follicles suggested a role in folliculogenesis. To study a presumed regulation of SNAP-25, we used a rat GC line (GFSHR-17), which expresses FSH receptors, and luteinizing human GC, which express LH receptors. FSH elevated SNAP-25 mRNA and protein levels about fivefold within 24 h in GFSHR-17 cells. The cAMP analogue dibutyryl-cAMP (db-cAMP) mimicked this action of FSH. The effects of both db-cAMP and FSH were inhibited by the protein kinase A (PKA) inhibitor H89. In contrast, SNAP-25 protein and mRNA-levels were not altered by LH/hCG in luteinized human GC. Our results for the first time identify SNAP-25b in oocytes and SNAP-25a in steroidogenic cells of the mammalian ovary. SNAP-25a and b may be involved in different exocytotic processes in these cell types.
Grosse
J
J
Anatomisches Institut, Technische Universität München, D-80802 München, Germany.
Bulling
A
A
Brucker
C
C
Berg
U
U
Amsterdam
A
A
Mayerhofer
A
A
Gratzl
M
M
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006062:Gonadotropins / Q000494:pharmacology*; D006801:Humans; D007986:Luteinizing Hormone / Q000494:pharmacology; D008565:Membrane Proteins; D009412:Nerve Fibers / Q000737:chemistry; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics*; D009865:Oocytes / Q000737:chemistry*; D010053:Ovary / Q000737:chemistry* / Q000294:innervation; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D050825:Synaptosomal-Associated Protein 25; D014018:Tissue Distribution
D006063:Chorionic Gonadotropin; D004791:Enzyme Inhibitors; D006062:Gonadotropins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C494208:SNAP25 protein, human; C494210:Snap25 protein, rat; D050825:Synaptosomal-Associated Protein 25; D003994:Bucladesine; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D017868:Cyclic AMP-Dependent Protein Kinases