dinoprost and calmidazolium

The aim of this study was to clarify the possible involvement of nitric oxide (NO) on prostaglandin (PG) E2-9-ketoreductase activity in the gonadotropin-releasing hormone (GnRH)-dependent PGF2 alpha synthesis by the interrenal gland of the female water frog, Rana esculenta, during the post-reproduction. Interrenal glands were incubated in vitro with GnRH, NO donor (sodium nitroprusside, SNP), and inhibitors of phospholipase C (compound 48/80), inositol triphosphate (decavanadate), calmodulin (calmidazolium), NO synthase (L-NAME), and PGE2-9-ketoreductase (palmitic acid). Production of PGE2 and PGF2 alpha and NO synthase and PGE2-9-ketoreductase activities were determined. GnRH and SNP increased PGF2 alpha production and PGE2-9-ketoreductase activity, and decreased production of PGE2 and GnRH increased NO synthase activity. GnRH effects were blocked by all inhibitors, except for palmitic acid, which did not affect NO synthase activity, which is increased by GnRH. This study indicates that NO may be involved in regulation of the R. esculenta post-reproduction through stimulation of PGE2-9-ketoreductase activity in GnRH-dependent PGF2 alpha synthesis by the frog interrenal gland.

Topics: Animals; Dinoprost; Dose-Response Relationship, Drug; Female; Gonadotropin-Releasing Hormone; Hydroxyprostaglandin Dehydrogenases; Imidazoles; In Vitro Techniques; Interrenal Gland; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase; Nitroprusside; p-Methoxy-N-methylphenethylamine; Palmitic Acid; Rana esculenta; Reproduction; Signal Transduction; Type C Phospholipases; Vanadates

The inhibition of prostaglandin (PG) synthesis found in early human decidua is antagonized by the anti-progestin, ZK 98734. This action of ZK 98734 is abolished by actinomycin, an inhibitor of protein synthesis and by the calcium channel blocker, verapamil. Calmidazolium, an antagonist of the intracellular calcium binding protein calmodulin was less effective in inhibiting the stimulation of PG synthesis induced by the anti-progestin. Chronic stimulation of protein kinase C activity by 1-oleoyl-2-acetyl-sn-glycerol (OAG) induced a slight reduction of PG release and was antagonized by polymixin. These findings suggest that inhibition of PG synthesis in early pregnancy is caused by progesterone and that increased release of PGs induced by anti-progestins is dependent on new protein formation and requires extracellular calcium.

Topics: Abortion, Spontaneous; Calmodulin; Cells, Cultured; Dactinomycin; Decidua; Diglycerides; Dinoprost; Estrenes; Female; Humans; Imidazoles; Kinetics; Pregnancy; Progesterone; Progestins; Radioimmunoassay; Verapamil

The pharmacological and biochemical mechanisms of contractile responses to the protein kinase C (PKC) activator phorbol-12,13-diacetate (PDA) were investigated in canine basilar arteries. In the normal medium, PDA elicited a strong, dose-related, and slow-developing sustained contraction. Among the constrictors examined, including serotonin, prostaglandin F2 alpha, and endothelin, only PDA yielded contractions in a Ca2(+)-free medium. In both media, the PDA-induced contractions were virtually inhibited by either staurosporine, H-7, or quinacrine, while neither neurotransmitter blockades nor R24571 (calmidazolium) exerted significant effects. In addition, it was shown that 8-bromocyclic GMP, but not 8-bromocyclic AMP, markedly curtailed the PDA-induced contractions. Biochemical analysis, furthermore, showed that PDA induced increased phosphorylations of 27- and 96-kDa and proteins other than the myosin light chain (MLC) 20-kDa protein. Thus, the present results open up a novel mechanism of sustained cerebral artery contractions, where PKC activation rather than Ca2+/calmodulin/MLC system plays a key role that is regulated both by phospholipase A2 and by cyclic GMP.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Alkaloids; Animals; Basilar Artery; Cyclic GMP; Dinoprost; Dogs; Endothelins; Enzyme Activation; Female; Imidazoles; Isoquinolines; Male; Muscle Contraction; Phorbol Esters; Phosphorylation; Piperazines; Protein Kinase C; Quinacrine; Serotonin; Staurosporine

The release of the prostanoids prostaglandin D2 (PGD2), prostaglandin E2 (PGE2) and thromboxane induced by zymosan and phorbol ester in cultured rat Kupffer cells was found to depend on the extracellular concentration of Ca2+ to some extent. Prostanoid formation following the addition of the calcium ionophore A 23187 was totally inhibited when calcium ions were withdrawn from the medium whereas the prostanoid synthesis from added arachidonic acid was independent of Ca2+. A half-maximal rate of PGE2 release by cells treated with zymosan, phorbol ester or A23187 was obtained at 0.6-0.7 microM free extracellular Ca2+ and greater than or equal to 100 microM free Ca2+ was required to stimulate PGE2 formation maximally. The calmodulin antagonist R24571 partially inhibited the release of PGE2 elicited by zymosan and A23187 but not by phorbol ester or arachidonic acid. Verapamil and nifedipine, two calcium channel blockers, had no effect on the formation of PGE2 irrespective of the stimulus. TMB 8 [3,4,5-trimethoxybenzoic acid 8-(diethylamino)-octyl ester] an intracellular calcium antagonist, inhibited the synthesis of PGE2 induced by zymosan and phorbol ester. The superoxide formation following the addition of zymosan and phorbol ester was not influenced by removal of calcium ions from the medium or by addition of the various calcium antagonists. The data presented here suggest that Ca2+-dependent reactions are involved in the synthesis of prostanoids induced by zymosan and phorbol ester and that both extracellular Ca2+ and mobilization of Ca2+ from intracellular stores are needed to induce maximally the production of prostanoids in cultured rat Kupffer cells.

Topics: Animals; Calcium; Calcium Channel Blockers; Calmodulin; Cells, Cultured; Dinoprost; Gallic Acid; Imidazoles; Kupffer Cells; Phorbol Esters; Prostaglandin D2; Rats; Superoxides; Zymosan

Plasma membrane fractions were prepared from ovaries of superovulated rats and examined for structural changes during luteolysis. Using fluorescence polarization, we observed a rapid rigidification in vitro of samples obtained from ovaries undergoing spontaneous or prostaglandin F2 alpha-induced regression. The rigidification, manifested by a 72% polarization increase over 50 min, is calcium and calmodulin dependent, temperature sensitive and protein mediated. This increase in polarization did not appear in fractions from nonregressing ovaries; however, addition of phospholipase A2 caused virtually identical changes in polarization results as in samples prepared from regressing ovaries. These results suggest that calcium-calmodulin-dependent phospholipase A2 plays a role in membrane deterioration during luteolysis.

Topics: Animals; Calcium; Calmodulin; Cell Membrane; Dinoprost; Female; Fluorescence Polarization; Imidazoles; Luteolysis; Phospholipases A; Phospholipases A2; Prostaglandins F; Rats; Rats, Inbred Strains; Temperature