Differential actions of phospholipase C on gonadotropin-releasing-hormone-stimulated release and glycosylation of luteinizing hormone in rat anterior pituitary cells.
Receptor-mediated activation of phospholipase C (PLC) which releases diacylglycerol and inositol trisphosphate has been implicated in the action of gonadotropin-releasing hormone (GnRH) on gonadotrophs. Previously we demonstrated that the synthetic diacylglycerol, phorbol 12-myristate 13-acetate (PMA) and PLC mimic the stimulatory effects of GnRH on both luteinizing hormone (LH) glycosylation and release. In this study we further investigated how PMA or PLC interact with GnRH to control LH release versus glycosylation. Cultured pituitary cells were incubated in the presence of radiolabeled precursors and GnRH (0, 1, or 100 nM), with or without PMA (10 nM) or PLC (0.24 U/ml) for 4 h. LH translation and glycosylation were monitored by measuring incorporation of [14C]alanine and [3H]glucosamine, respectively, into total (cell and medium) immunoprecipitable LH. Immunoreactive LH (IRLH) was measured by radioimmunoassay. Both PMA and PLC increased (p < 0.01) basal IRLH release, and IRLH release stimulated by 1 nM GnRH. Neither PMA nor PLC exerted an additive effect on IRLH release stimulated by 100 nM GnRH. The interactions between PMA or PLC and GnRH on IRLH release were significant (p < 0.01). Both PMA and PLC elevated (p < 0.01) total [3H]glucosamine-LH, but had no additive effect with 1 nM GnRH; PLC depressed (p < 0.05) the stimulatory effect of 100 nM GnRH, whereas PMA had no effect. The interactions between PMA or PLC and GnRH on LH glycosylation were significant (p < 0.01). PMA, PLC or GnRH alone did not affect total [14C]alanine-LH. In the presence of 1 or 100 nM GnRH, PLC, but not PMA, decreased (p < 0.05) total [14C]alanine-LH.(ABSTRACT TRUNCATED AT 250 WORDS)
Liu
T C
TC
Department of Medical Technology, National Yang-Ming Medical College, Taipei, Taiwan, Republic of China.
Pu
H F
HF
Jackson
G L
GL
D000409:Alanine / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005944:Glucosamine / Q000378:metabolism; D006031:Glycosylation; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology*; D007986:Luteinizing Hormone / Q000378:metabolism*; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism / Q000494:pharmacology*
D007987:Gonadotropin-Releasing Hormone; D007986:Luteinizing Hormone; D010738:Type C Phospholipases; D005944:Glucosamine; D013755:Tetradecanoylphorbol Acetate; D000409:Alanine