Rab3B is essential for GnRH-induced gonadotrophin release from anterior pituitary cells.
Gonadotrophin-releasing hormone (GnRH) induces the release of gonadotrophins via an increase in cytosolic Ca2+ concentration ([Ca2+]). Rab3B, a member of the small GTP-binding protein Rab family, is known to be involved in Ca(2+)-regulated exocytosis in pituitary cells. However, it is not known whether Rab3B functions in the physiological process regulated by GnRH in gonadotrophs. In this study using antisense oligonucleotide against Rab3B (AS-Rab3B) we determined that Rab3B is involved in GnRH-induced gonadotrophin release. Rab3B immunopositive cells were reduced in 24% of pituitary cells by AS-Rab3B. This treatment did not affect the population of gonadotrophs or the intracellular contents of gonadotrophins. However, AS-Rab3B significantly inhibited the total amount of basal and GnRH-induced gonadotrophin released from pituitary cells. These results show that Rab3B is involved in basal and GnRH-induced gonadotrophins release but not the storage of gonadotrophins. Next, the changes in [Ca2+] and exocytosis in gonadotrophs treated with AS-Rab3B were compared among Rab3B-positive and -negative cells. The change in [Ca2+] was not different in the two groups, but exocytosis was significantly inhibited in Rab3B-negative cells. These results suggest that Rab3B is essential for GnRH-regulated exocytosis downstream of cytosolic Ca2+ in gonadotrophs.
Tasaka
K
K
Department of Obstetrics and Gynecology, Osaka University Medical School, Japan.
Masumoto
N
N
Mizuki
J
J
Ikebuchi
Y
Y
Ohmichi
M
M
Kurachi
H
H
Miyake
A
A
Murata
Y
Y
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D005089:Exocytosis / Q000187:drug effects; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000502:physiology*; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology*; D006065:Gonadotropins, Pituitary / Q000378:metabolism*; D015151:Immunoblotting; D007986:Luteinizing Hormone / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D020692:rab3 GTP-Binding Proteins
D006065:Gonadotropins, Pituitary; D016376:Oligonucleotides, Antisense; D007987:Gonadotropin-Releasing Hormone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D019204:GTP-Binding Proteins; D020692:rab3 GTP-Binding Proteins; D002118:Calcium