ABSTRACT
The present study was attempted to examine the effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on catecholamine (CA) secretion evoked by cholinergic stimulation, membrane depolarization and calcium mobilization from the isolated perfused rat adrenal gland. The perfusion of PACAP (10 nM) into an adrenal vein for 60 min produced a great inhibition in CA secretion evoked by ACh (5.32 X 10(-3) M), high K+ (5.6 X 10(-2) M), DMPP (10(-4) M for 2 min), McN-A-343 (10(-4) M for 2 min), cyclopiazonic acid (10(-5) M for 4 min) and Bay-K-8644 (10(-5) M for 4 min). Also, in the presence of neuropeptide (NPY), which is known to be co-localized with norepinephrine in peripheral sympathetic nerves, CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were also significantly depressed. However, in adrenal glands preloaded with PACAP (10 nM) under the presence of VIP antagonist ((Lys1, Pro2.5, Arg3.4, Tyr6)-VIP (3 micrometer)) for 20 min, CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were not altered greatly in comparison to the case of PACAP-treatment only. Taken together, these results suggest that PACAP causes the marked inhibition of CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by membrane depolarization, indicating that this effect may be mediated by inhibiting influx of extracellular calcium and release in intracellular calcium in the rat adrenomedullary chromaffin cells.
Subject(s)
Animals , Rats , (4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethylammonium Chloride , 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester , Adrenal Glands , Calcium , Chromaffin Cells , Dimethylphenylpiperazinium Iodide , Membranes , Neuropeptides , Norepinephrine , Perfusion , Pituitary Adenylate Cyclase-Activating Polypeptide , Potassium , VeinsABSTRACT
BACKGROUND: The influence of gamma-aminobutyric acid(GABA), which is well-known as a major inhibitory neurotransmitter in central nervous system, on secretion of catecholamines(CA) was investigated in the isolated perfused rat adrenal gland. METHODS: Mature male Sprague-Dawley rats were anesthetized with ether. Ther adrenal gland was isolated by the methods f Wakade. A cannula used for perfusion of the adrenal gladn was inserted into the distal end of the renal vein. The adrenal gland, along with ligated blood vessels and the cannula, was carefully removed from the animal and placed on a platform of a leucite chamber. RESULTS: GABA given into an adrenal vein of the rat produced markedly secretion of CA from the adrenal gland. Tachyphylaxis to the relesing effect of CA evoked by GABA was observed. The secretory effect of CA evoked by GABA was attenuated singnificantly by pretreatment with mecamylamine or atropine. Ouabain inhibited greatly the secretory response of GABA. When omitting the external potassium ion, the basal release of CA was increased. During this period GABA no longer revealed the increase in CA release. CA secretion evoked by GABA was blocked significantly by perfusion of calcium-free Krebs solution containing 5mMEGTA for 30-min. Pretreatment with bicuculline or picrotoxin inhibited CA secretion evoked by GABA as well as ACh. ACh-evoked CA release was potentiated by GABA infusion(400ug/30min). CONCLUSION: The experimental findings suggest that GABA causes the secretory effect of CA in a fashion of external calcium and potassium iosn-dependence, and that this releasing effect of CA induced by GABA may be exterted by stimulation of GABAergic A-reccptors located on adrenomedullary chromaffine cell, which is likely associated with cholinergic receptor activation evoked CA secretion.