title: Involvement of calcium, inositol-1,4,5 trisphosphate and diacylglycerol in the prothoracicotropic hormone-stimulated ecdysteroid synthesis and secretion in the prothoracic glands of Bombyx mori.
abstract: The objective of this study was to determine which intracellular second messenger systems are activated by prothoracicotropic hormone in the prothoracic glands (PGs) of Bombyx mori. Recombinant prothoracicotropic hormone (rPTTH) could stimulate ecdysteroid synthesis and secretion from day 6 PGs of the 5th instar of Bombyx mori within 30 min of in vitro incubation. However, rPTTH did not stimulate any increases in the glandular content of inositol 1,4,5-trisphosphate and cAMP during this short incubation period. Extracellular Ca2+ influenced the basal and rPTTH-stimulated ecdysteroid synthesis and release in a dose-dependent manner. The L-type Ca2+ channel antagonist, nitrendipine, inhibited the rPTTH-stimulated ecdysteroid synthesis and secretion (IC50-28 microM). The phospholipase C inhibitor, 2-nitro-4-carboxyphenyl-N, N-diphenylcarbamate, inhibited the rPTTH-stimulated ecdysteroid synthesis (IC50-19 microM). The protein kinase C inhibitor, chelerythrine chloride, inhibited the rPTTH-stimulated ecdysteroid synthesis (IC50-14 microM). The protein kinase C activator, phorbol-12-myristate 13-acetate (PMA), could stimulate basal ecdysteroid synthesis and secretion (EC50-1 microM) and its inactive alpha-isomer (4 alpha-PMA) was ineffective. The combined results suggest that the PTTH-stimulated ecdysteroid synthesis and release in the PGs of Bombyx is dependent on extracellular Ca2+ and the bifurcating second messenger signalling cascade of inositol 1,4,5-triphosphate and diacylglycerol.
authors.affiliation: Institute of Biology, National Center for Scientific Research Demokritos, Aghia Paraskevi, Athens 15310, Greece.
authors.forename: S G
authors.initials: SG
authors.lastname: Dedos
authors.forename: H
authors.initials: H
authors.lastname: Fugo
mesh_terms: D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D012831:Bombyx / Q000502:physiology*; D002118:Calcium / Q000494:pharmacology / Q000502:physiology*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002219:Carbamates / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004075:Diglycerides / Q000494:pharmacology / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D026461:Ecdysteroids / Q000096:biosynthesis / Q000378:metabolism; D004702:Endocrine Glands / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology / Q000502:physiology*; D007301:Insect Hormones / Q000494:pharmacology / Q000502:physiology*; D009568:Nitrendipine / Q000494:pharmacology; D010617:Phenanthridines / Q000494:pharmacology; D048448:Phenylcarbamates; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011994:Recombinant Proteins / Q000494:pharmacology; D015290:Second Messenger Systems / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors
chemical_list: D000470:Alkaloids; D053119:Benzophenanthridines; D002121:Calcium Channel Blockers; D002219:Carbamates; D004075:Diglycerides; D026461:Ecdysteroids; D004791:Enzyme Inhibitors; D007301:Insect Hormones; D010617:Phenanthridines; D048448:Phenylcarbamates; D011994:Recombinant Proteins; C025508:2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate; C009089:prothoracicotropic hormone; D015544:Inositol 1,4,5-Trisphosphate; D009568:Nitrendipine; D000242:Cyclic AMP; C016299:chelerythrine; D011493:Protein Kinase C; D010738:Type C Phospholipases; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium