store acetylcholine (ACh) ( Fritz et al. 1999 ). Both in vivo and in vitro experiments showed that human GCs also express several muscarinic (MR) subtype receptors ( Mayerhofer & Fritz 2002 , Cruz et al. 2015 ). Finally, the enzyme that degrades
Raul Riquelme, Freddy Ruz, Artur Mayerhofer and Hernán E Lara
Matthias R Meyer, Natalie C Fredette, Matthias Barton and Eric R Prossnitz
factors, such as nitric oxide (NO), and contracting factors, such as cyclooxygenase (COX)-derived vasoconstrictor prostanoids and endothelin-1 ( Feletou & Vanhoutte 2006 ). Studies on endothelial function widely rely on acetylcholine as a muscarinic
Yusuke Seino, Takashi Miki, Wakako Fujimoto, Eun Young Lee, Yoshihisa Takahashi, Kohtaro Minami, Yutaka Oiso and Susumu Seino
.2 −/− mice, GIIS induced by GLP-1 is markedly attenuated when compared with that in the Kir6.2 + / + mice ( Fujimoto et al . 2009 ). In addition, acetylcholine stimulation was shown to increase intracellular Ca 2 + concentrations ([Ca 2 + ] i ) in β
A. Shainberg, H. Brik, R. Bar-Shavit and S. R. Sampson
Studies were made on the effect of thyroid hormones on the level of acetylcholine receptors (AChR) in cultured rat skeletal muscle. Treatment of differentiated myotubes in vitro with thyroxine (T4; 2 × 10−7 mol/l) for 2–3 days caused a marked decrease in the amount of AChR (P<0·05) and an increase in activity of Na+-K+-ATPase (P<0·05). There was no significant effect of hormone treatment on other muscle proteins, such as creatine kinase and acetylcholinesterase. Measurements of the turnover rate of AChR in T4-treated myotubes showed only a very slight effect of T4 on the rate of AChR degradation. To study the mechanism by which the hormone exerts its effect, muscle cells were labelled with radioactive amino acid and the rate of its incorporation into AChR protein was measured. The AChR was then isolated using anti-AChR antibodies. The specific activity of labelled AChR was lower in hormone-treated cells. These experiments suggest that the decreased level of AChR in response to thyroid hormone treatment is due to a partial suppression of receptor synthesis.
J. Endocr. (1984) 101, 141–147
JOAN PICKENS and MARY F. LOCKETT
A concentration of 0·01 μg. (—)-triiodothyronine in the Tyrode's fluid bathing a phrenic nerve diaphragm preparation has been shown to decrease the acetylcholine appearing in the bath fluid in response to tetanus of the diaphragm through the phrenic nerve at a fixed number of impulses per sec. for a fixed number of minutes, when (a) eserine (40 μg./ml.) was used to inhibit the cholinesterase and the acetylcholine was assayed on leech muscle; and (b) prostigmine (5 × 10-6) was used as the anticholinesterase and the acetylcholine was assayed by its depressor effect on the arterial pressure of rats.
M. Benyamina, F. Leboulenger, I. Lirhmann, C. Delarue, M. Feuilloley and H. Vaudry
The effect of cholinergic agonists on glucocorticoid and mineralocorticoid production by frog interrenal (adrenal) tissue was studied in vitro by means of continuous perifusion. Acetylcholine, at doses ranging from 1 to 100 μmol/l, stimulated both corticosterone and aldosterone output in a dose-dependent manner, with a half-maximal effective dose of 2·5 μmol/l. Corticosteroid production was also stimulated by muscarine (10 μmol/l). In contrast, neither nicotine nor nicotine bitartrate (1–100 μmol/l) enhanced corticosteroid biosynthesis. The kinetics of the response of adrenal cells to acetylcholine and muscarine were similar to those observed during angiotensin II stimulation. In particular, a significant reduction (20–40%) in the spontaneous level of corticosteroid production was recorded after the initial infusion of muscarinic agents, but no further decrease in the basal level occurred after a second cholinergic administration. The effect of acetylcholine was blocked by the muscarinic receptor antagonist atropine (10 μmol/l). These results indicate that acetylcholine can stimulate frog adrenocortical cells through muscarinic receptors. Repeated 20-min pulses of acetylcholine (50 μmol/l) or muscarine (10 μmol/l), given at one pulse per 130 min, resulted in a marked reduction in the secretory response to the second pulse. No reduction in the stimulatory effect of acetylcholine or muscarine was observed when a 6·5-h interval separated two 20-min infusions of the secretagogue. In contrast with these findings, iterative pulses of the muscarinic agonist pilocarpine (in the range 1–100 μmol/l) did not cause any desensitization.
These data show that the neurotransmitter acetylcholine can modulate frog adrenocortical function and suggest that, in addition to more conventional regulators, i.e. ACTH and angiotensin II, the cholinergic endings of the splanchnic nerve might participate in the regulation of corticosteroid secretion, at least under some physiological conditions such as neurogenic stress.
J. Endocr. (1987) 113, 339–348
K. YAMASHITA, M. MIENO, T. SHIMIZU and ER. YAMASHITA
The effect of prostaglandin E2 (PGE2) on the secretion of adrenaline and noradrenaline by the adrenal gland and the interaction between PGE2 and acetylcholine in the adrenal medulla were examined in anaesthetized dogs. In splanchnicotomized dogs, i.v. injection of PGE2 failed to induce any secretion of catecholamines from the adrenal gland, whereas administration of PGE2 into the lumboadrenal artery resulted in a slight, approximately dose-dependent increase in catecholamine secretion within 2 min of the injection. This effect of PGE2 was unaffected by i.v. administration of atropine. Intravenous administration of acetylcholine 1 min after the administration of PGE2 into the lumboadrenal artery of splanchnicotomized atropine-treated dogs had a markedly greater effect on adrenal catecholamine secretion; the resultant output was about twice that evoked by acetylcholine in the absence of PGE2. The effect was more than additive, since the response to acetylcholine was at least one order of magnitude greater than that to PGE2. This indicates that PGE2 and acetylcholine may act synergistically in the adrenal medulla.
M. Feuilloley, P. Netchitaïlo, C. Delarue, F. Leboulenger, M. Benyamina, G. Pelletier and H. Vaudry
In order to determine the role of the cytoskeleton in adrenal steroidogenesis, we have studied the effect of cytochalasin B (a microfilament-disrupting agent) and vinblastine (an antimicrotubular drug) on corticosteroid secretion by frog interrenal tissue in vitro. Perifusion of interrenal fragments with cytochalasin B (50 μmol/l) induced a marked inhibition of basal corticosteroid output. In addition, stimulation of corticosteroidogenesis by all corticotrophic factors was also inhibited by cytochalasin B. Using an immunohistochemical technique and specific anti-tubulin antiserum, we verified that vinblastine (10 μmol/l) was responsible for the disappearance of the microtubular network in adrenocortical cells. Administration of vinblastine (10 μmol/l) did not affect the spontaneous secretion of corticosterone and aldosterone and had no effect on the steroidogenic response of interrenal glands to angiotensin II and acetylcholine. In contrast, vinblastine was responsible for a marked decrease in serotonin-induced stimulation of corticosteroid production. On the other hand, data from high-performance liquid chromatography showed that infusion of cytochalasin B or vinblastine was not associated with the production of any new steroid which could interfere in the radioimmunoassays. Taken together, these data suggest that microfilaments are involved in a late and common step of corticosteroidogenesis while microtubules are only required for the coupling of the secretory response to certain corticotrophic factors such as ACTH and serotonin.
J. Endocr. (1988) 118, 365–374
P. W. YOUNG, R. J. BICKNELL and J. G. SCHOFIELD
Acetylcholine (25 μmol/l) in the presence of the choline esterase inhibitor physostigmine (67 μmol/l) increased the release of growth hormone and efflux of 45Ca2+ from perifused bovine pituitary slices; the time taken for the maximal response to occur was the same. In batch incubations, acetylcholine (1 μmol/l–1 mmol/l) increased pituitary cyclic GMP concentrations in the pituitary gland within 2 min, and increased incorporation of [3H]inositol and [32P]phosphate into pituitary phosphatidyl inositol within 15 min. Cyclic AMP concentrations were not significantly changed 2 or 5 min after acetylcholine addition. All the tissue responses were inhibited by prior exposure of the tissue to atropine (1 μmol/l) but not by tubocurarine (10 μmol/l–1 mmol/l), indicating that the responses were mediated by receptors of the muscarinic type. The similarities between these responses and those to known hypothalamic hypophysiotrophic hormones are discussed.
J. G. Schofield, A. I. Khan and A. Wood
Acetylcholine is known to stimulate the secretion of growth hormone and prolactin and the efflux of 86Rb from bovine anterior pituitary cells: dopamine prevents the stimulation of 86Rb efflux and of prolactin but not growth hormone secretion. The sensitivity of these responses to pertussis toxin has been determined.
Treatment of bovine anterior pituitary cells in primary culture with pertussis toxin (18 h, 100 ng/ml) did not modify the stimulation of prolactin secretion by acetylcholine, but prevented its inhibition by dopamine. In lactotrophs, dopamine but not acetylcholine receptors are therefore coupled to secretion through a pertussis toxin substrate. The stimulation of 86Rb efflux by acetylcholine was also unaffected by pertussis toxin and, again, its inhibition by dopamine was prevented.
Treatment of the cells with pertussis toxin enhanced the secretion of growth hormone in response to acetylcholine. Nitrendepine (1 μmol/l) prevented the cholinergic stimulation of growth hormone but not prolactin secretion from these cells. Acetylcholine increased the cytoplasmic calcium concentration and this rise was enhanced by treatment of the cells with pertussis toxin. Nitrendepine partially inhibited the rise in calcium caused by acetylcholine, and prevented the enhancement of the rise following pertussis toxin treatment.
Cholinergic stimulation of growth hormone therefore depends on calcium entry through nitrendepine-sensitive channels, whereas stimulation of prolactin secretion does not, and in somatotrophs a pertussis toxin substrate may limit calcium entry through these channels. These different sensitivities of somatotrophs and lactotrophs to pertussis toxin and nitrendepine may reflect differences in the properties of the predominant calcium currents in the two cell types.
J. Endocr. (1988) 116, 393–401