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Biomedical subjects

B G Livett

Publications and source records attributed to B G Livett.

At least 73 records · Page 4Linked to original sources
PubMed1986

Effects of opioid peptides containing the sequence of Met5-enkephalin or Leu5-enkephalin on nicotine-induced secretion from bovine adrenal chromaffin cells.

Eighteen endogenous opioid peptides, all containing the sequence of either Met5- or Leu5-enkephalin, were tested for their ability to modify nicotine-induced secretion from bovine adrenal chromaffin cells. ATP released from suspensions of freshly isolated cells was measured with the luciferin-luciferase bioluminescence method as an index of secretion. None of the peptides affected 5 microM nicotine-induced ATP release at 10 nM. Three peptides inhibited secretion at 5 microM: dynorphin1-13, dynorphin1-9, and rimorphin inhibited by 65%, 37%, and 29% respectively. Use of peptidase inhibitors (bestatin, thiorphan, bacitracin, or 1,10-phenanthroline) did not result in any of the other peptides showing potent actions on the nicotinic response, although bestatin and thiorphan did enhance the inhibitory actions of dynorphin1-13 and dynorphin1-9 by 20-30%. Nicotine-induced secretion of endogenous catecholamines from bovine chromaffin cells cultured for 3 days was also studied to assess any selective actions of the peptides on adrenaline or noradrenaline cell types. Dynorphin1-13 was 1,000-fold more potent than Leu5-enkephalin at inhibiting endogenous catecholamine secretion. Dynorphin1-13 was slightly more potent at inhibiting noradrenaline release than adrenaline release whereas Leu5-enkephalin showed the opposite selectivity. The structure-activity relationships of opioid peptide actions on the chromaffin cell nicotinic response are discussed in relation to the properties of the adrenal opioid binding sites.

Adenosine Triphosphate↗
PubMed1986

Effects of opioid peptides and morphine on histamine-induced catecholamine secretion from cultured, bovine adrenal chromaffin cells.

The effect of opioid peptides and morphine on histamine-induced catecholamine secretion has been studied in monolayer cultures of dispersed, bovine adrenal chromaffin cells. Histamine-induced a dose-dependent secretion of both adrenaline and noradrenaline with a threshold dose of approximately 5 nM, an EC50 of 150 nM and maximal secretion at 10 microM. Catecholamine secretion induced by 1 microM histamine was completely dependent on extracellular calcium, was inhibited in a dose-dependent manner by mepyramine (1 nM-1 microM), and was unaffected by cimetidine (10 microM) and hexamethonium (0.1 mM). Dynorphin-1-13 (1 nM-20 microM), metorphamide (0.1 nM-10 microM), morphine (1 nM-0.1 mM) and diprenorphine (1 nM-0.1 mM) each had no effect on adrenaline or noradrenaline secretion induced by 1 microM histamine. The characteristics of histamine-induced catecholamine secretion from bovine adrenal chromaffin cells were similar to those reported previously for cat and rat adrenal medulla being calcium-dependent and mediated by H1 histamine-receptors. The results with opioid peptides and morphine suggest that endogenous adrenal opioid peptides do not act on the opioid binding sites found on adrenal medullary chromaffin cells to modify their secretory response to histamine.

Adrenal Glands↗
PubMed1986

The role of sensory fibres in the rat splanchnic nerve in the regulation of adrenal medullary secretion during stress.

We have studied the involvement of sensory nerves containing substance P (SP) in the modulation of stress-induced catecholamine (CA) secretion from the sympathetic nervous system and adrenal medulla. Adrenaline and noradrenaline (NA) levels were measured in blood samples withdrawn from the inferior vena cava (i.v.c.) at 5 or 15 min intervals for periods of up to 60 min, in adult rats during stress induced by insulin or cold. Insulin stress caused a biphasic elevation of plasma CA. Previous studies from our laboratory have shown that the first phase lasting 30 min is neurogenic, and the second phase from 30 to 60 min is non-neurogenic in mechanism. In control adult rats (with normal levels of SP in their splanchnic nerve), insulin stress caused a slow and progressive secretion of adrenaline into the circulation for the first 30 min (neurogenic phase). In the period 30-60 min (non-neurogenic phase) plasma adrenaline and NA levels rose at a much higher rate. In capsaicin-pre-treated rats (in which SP levels in the splanchnic nerve were depleted by 68%) insulin stress produced a steady increase in plasma adrenaline levels for up to 5 min similar to that in insulin-stressed control animals; however, by 10 min the plasma adrenaline levels had fallen to basal and remained low up to 30 min. From 30 to 60 min, plasma adrenaline and NA levels rose steeply as seen with control animals. We conclude that capsaicin pre-treatment affected the neurogenic phase but did not affect the non-neurogenic phase. Cold stress increased the plasma adrenaline levels by a neurogenic mechanism over 30 min in control rats. In contrast, in capsaicin-pre-treated, cold-stressed rats, plasma adrenaline did not increase significantly. Plasma NA levels were also significantly lowered in capsaicin-pre-treated, cold-stressed rats during the neurogenic phase but NA increases were not dependent on an intact adrenal innervation. The results using both insulin stress and cold stress suggest that capsaicin-sensitive (sensory) nerve fibres in the adrenal medulla and in sympathetic ganglia are capable of modifying the secretory responses of these tissues to stress. Results from our previous in vitro work are compatible with the view that SP may be the neuromodulator released from such sensory nerves to produce these effects. This suggests that the previously reported ability of SP to modulate nicotinic receptor function in vitro by either inhibiting the nicotinic response or protecting against nicotinic desensitization may be more than a mere pharmacological curiosity.(ABSTRACT TRUNCATED AT 400 WORDS)

Adrenal Medulla↗
PubMed1986

Elevation in plasma catecholamines in response to insulin stress is under both neuronal and nonneuronal control.

We have examined the contribution of neurogenic and nonneurogenic influences to the secretion of adrenal catecholamines (CA) in adult rats after iv administration of insulin. Plasma CA levels were measured in control rats and in rats after surgical or pharmacological adrenal denervation or adrenalectomy. Insulin-induced CA secretion was biphasic and proportional to the insulin dose used. The first phase was neurogenic in origin and produced a moderate increase in plasma adrenaline (A) levels, with little or no change in noradrenaline (NA) levels. This neurogenic increase in plasma A was reduced by partial denervation and abolished by surgical complete adrenal denervation, adrenalectomy, or administration of hexamethonium and atropine. The second phase occurred later and produced a dramatic increase in both plasma A and NA. This phase was initiated when the plasma glucose level fell below 75 mg/100 ml. This late release of A and NA was not altered by surgical or pharmacological adrenal denervation, showing that it was nonneuronal in origin. However, both the early and late phases were abolished by adrenalectomy, showing that adrenal secretion of CA was the origin of the increased plasma levels of NA and A. The late rise in plasma CA was also abolished by iv administration of glucose. These data suggest that the mechanism responsible for the nonneurogenic secretion of adrenal CA in response to insulin stress was sensitive to the level of hypoglycemia.

Adrenal Glands↗
PubMed1985

The substance P receptor subtype modulating catecholamine release from adrenal chromaffin cells.

Substance P (SP) has two distinct actions on catecholamine (CA) release from cultured bovine adrenal chromaffin cells: SP inhibits acetylcholine (ACh)- or nicotine-induced [3H]norepinephrine ([3H]NE) release; and SP protects against desensitization of ACh- or nicotine-induced [3H]NE release. In order to characterize the SP receptor subtypes modulating these two actions, we have tested several SP-related tachykinins and SP analogues for their ability to act as agonists of SP for these actions. The naturally occurring tachykinins, physalaemin, eledoisin and kassinin, were about equipotent and all much less potent than SP in inhibiting nicotine-induced [3H]NE release. Physalaemin and eledoisin were also approximately equipotent and much less potent than SP in protecting against desensitization of nicotine-induced [3H]NE release; kassinin slightly enhanced nicotinic desensitization. These results indicate that neither a kassinin-like nor a physalaemin-like peptide are likely to modulate adrenal CA release. The 'SP-P' receptor-selective analogue, SP-methyl ester, was an agonist, equipotent with SP, for both of SP's actions on [3H]NE release. [D-Pro2,D-Trp7,9]-SP, a potent antagonist of SP's actions in many SP-responsive tissues, was inactive as an antagonist of both of SP's actions on [3H]NE release from adrenal chromaffin cells; however, this analogue was a potent SP agonist in chromaffin cells. The rank order of potency of these and other SP-related peptides for inhibition of [3H]NE release was similar to their order of potency for protection against desensitization. Thus the chromaffin cell SP receptors mediating inhibition of nicotine-induced [3H]NE release appear to be very similar to those mediating protection against desensitization of nicotine-induced [3H]NE release. The results indicate that the SP receptors modulating adrenal CA release are different from either the 'SP-P' or 'SP-E' receptor subtypes; bovine adrenal chromaffin cells appear to possess a third subtype of SP receptor which shares similar characteristics with the SP receptors mediating histamine release from rat mast cells.

Adrenal Medulla↗
PubMed1985

Neuropeptides in the autonomic nervous system.

This review will concern itself with recent advances in our knowledge about the anatomy, cellular distribution, and proposed function of neuropeptides in the peripheral nervous system. In particular, the concept of co-storage and co-release of neuropeptides with conventional neurotransmitters will be dealt with in regard to their known distribution in the three broad divisions of the autonomic nervous system: sensory neurons, autonomic neurons, and the adrenal medulla. The proposed physiological functions of the peptides in these three systems will be reviewed.

Adrenal Medulla↗
PubMed1984

Neonatal capsaicin treatment prevents insulin-stress-induced adrenal catecholamine secretion in vivo: possible involvement of sensory nerves containing substance P.

We have investigated the effects of neonatal capsaicin treatment on stress-induced catecholamine release from adult rats in vivo. Catecholamines released into the inferior vena cava following insulin injection were separated by HPLC and measured by electrochemical detection. Neonatal capsaicin treatment depleted the substance P levels in the splanchnic nerve of adult rats by 70% and prevented the insulin-induced hypoglycaemia from evoking release of adrenaline. The results of this study provide the first evidence in vivo for the neuromodulatory role of sensory nerves containing substance P in the secretion of catecholamines from the adrenal medulla.

Adrenal Glands↗
PubMed1984

Desensitization to nicotinic cholinergic agonists and K+, agents that stimulate catecholamine secretion, in isolated adrenal chromaffin cells.

Desensitization of catecholamine (CA) release from cultured bovine adrenal chromaffin cells was studied to characterize the phenomenon of desensitization and to attempt an elucidation of the mechanism(s) involved in this phenomenon at the level of the isolated chromaffin cell. Prior exposure of chromaffin cells to nicotinic cholinergic agonists [acetylcholine (ACh) or nicotine] caused a subsequent depression or desensitization of CA release during restimulation of the cells with the same agonists. Rates of development of and recovery from nicotinic desensitization were in the minute time range and the magnitude of nicotinic desensitization of CA release was greater at 37 degrees C than at 23 degrees C. ACh- (or nicotine)-induced desensitization was shown to be the result of two processes: (1) a Ca2+-dependent component of desensitization, possibly due to a depletion of intracellular CA stores and (2) a Ca2+-independent, depletion-independent component of desensitization. Prior exposure of cultured chromaffin cells to an elevated concentration of K+ also resulted in desensitization of K+-induced CA release in these cells. K+-induced desensitization was completely Ca2+-dependent and was shown to be the result, at least in part, of a mechanism that is independent of depletion of CA stores.

Acetylcholine↗
PubMed1984

Substance P protects against desensitization of the nicotinic response in isolated adrenal chromaffin cells.

Substance P, a peptide endogenous to the splanchnic nerve, is known to inhibit the acetylcholine-and nicotine-induced release of catecholamines from isolated adrenal chromaffin cells. In the present study the effect of substance P on desensitization of catecholamine release from these cells was examined. Substance P (10(-5) M) completely protected against desensitization of catecholamine release produced by acetylcholine at 37 degrees C or 23 degrees C and by nicotine at 23 degrees C; substance P also afforded appreciable protection against nicotine-induced desensitization at 37 degrees C. The peptide had no effect on K+-induced desensitization of catecholamine release. Like substance P, d-tubocurarine also prevented nicotinic desensitization. Substance P prevented both of two components of nicotinic desensitization, i.e. the Ca2+-dependent component and the Ca2+-independent, depletion-independent component of desensitization. Substance P had little effect on subsequent catecholamine uptake, indicating that substance P's protection against desensitization is a result of facilitation of catecholamine release rather than inhibition of catecholamine reuptake. Nicotine-induced catecholamine release and nicotinic desensitization of catecholamine release were Na+-independent, although substance P's inhibition of nicotine-induced catecholamine release was reduced by extracellular Na+. These in vitro studies suggest a similar role for substance P in vivo: substance P's protection against nicotinic desensitization may ensure a maintained output of adrenal catecholamines during stress, when the splanchnic nerve releases large amounts of acetylcholine.

Acetylcholine↗
PubMed1984

Parallel but separate release of catecholamines and acetylcholinesterase from stimulated adrenal chromaffin cells in culture.

A pharmacological study was made of the effects of veratridine and lasalocid on the release of catecholamines, acetylcholinesterase (AChE) and dopamine-beta-hydroxylase (DBH) from cultures of isolated bovine adrenal chromaffin cells. Exposure of the cultures to veratridine resulted in concomitant release of catecholamines and AChE into the external medium in a dose-dependent and Ca2+-dependent manner. A Ca2+ ionophore, lasalocid, also produced a dose-dependent and parallel release of both catecholamines and AChE. The release of the two components was accompanied by release of DBH. The present results provide pharmacological evidence for a parallel release of catecholamines, AChE, and DBH from cultured adrenal chromaffin cells, and the stoichiometry of the release evoked by different secretagogues suggests that AChE and catecholamines are released from different cellular compartments.

Acetylcholinesterase↗
PubMed1984

Receptors and receptor modulation in cultured chromaffin cells.

The colocalization of acetylcholine (ACh) and neuropeptides (e.g., substance P and enkephalins) in the splanchnic nerve terminals suggests that these compounds might interact to modulate adrenal catecholamine release. Use has been made of primary monolayer and suspension cultures of bovine adrenal chromaffin cells to investigate postsynaptic receptor interactions between acetylcholine and a number of neuropeptides endogenous to the adrenal medulla and splanchnic nerve. The cells have both nicotinic and muscarinic acetylcholine receptors, but only the nicotinic receptors stimulate catecholamine release. Substance P, somatostatin, and the enkephalins all produced an inhibition of the ACh-evoked secretion of catecholamines, but their potency ranged over 100-fold. Substance P was the most potent with a mean inhibitory concentration (IC50) of 10(-6) M and Leu-enkephalin the least potent with an IC50 greater than 10(-4) M. These pharmacological effects were monitored conveniently by measuring the release of [3H]norepinephrine preloaded into the cells or alternatively, "on-line" by measuring ATP released into an incubation medium containing luciferin and firefly tail extract (luciferase). Of interest, the endogenous enkephalin heptapeptide (Met-enkephalin Arg6-Phe7) and "big" Met-enkephalin (BAM- 22P ) were some 100-fold more effective than Leu- or Met-enkephalin at inhibiting the nicotinic secretin of catecholamines, suggesting that a unique opiate receptor may be involved. Substance P had two distinct actions on the nicotinic response: (1) substance P inhibited acetylcholine-induced release of catecholamines; and (2) substance P protected against acetylcholine-induced desensitization of catecholamine release. With regard to (1), substance P inhibited the secretion of catecholamines and ATP evoked by acetylcholine or nicotine but not that evoked by K+ or veratridine, nor did substance P by itself affect secretion. Substance P appeared to interact with a regulatory site on the acetylcholine receptor - ionophore complex. Substance P receptors on chromaffin cells have similar structural requirements for activation as do substance P receptors in other substance P responsive tissues. With regard to (2), substance P (greater than 5 X 10(-6) M) completely protected against desensitization of catecholamine release produced by acetylcholine (greater than 10(-4) M) or nicotine (greater than 2.5 X 10(-6) M) with no effect on K+-induced desensitization.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗
PubMed1983

Use of isolated chromaffin cells to study basic release mechanisms.

An account is given of the authors' work with isolated adrenal chromaffin cells to study the synthesis, storage and release of catecholamines and of a number of neuropeptides endogenous to the adrenal medulla. A review of other studies in the literature with the isolated chromaffin cell system is included. It is seen that the isolated chromaffin cells are a convenient in vitro system well-suited to studies of basic release mechanisms. The isolated adrenal chromaffin cells maintain high levels of catecholamines and opiates and release them by exocytosis. The cells have both nicotinic and muscarinic receptors but only the nicotinic are involved in the agonist-evoked release of catecholamines (EC50 nicotine 5 X 10(-6) M: ACh 5 X 10(-5) M). The cells can synthesize AChE and selectively release the 10S molecular form by a mechanism different from exocytosis. Substance P (SP) modulates the secretion of catecholamines and ATP evoked by ACh or nicotine but not that evoked by K+ or veratridine. SP appears to interact with the nicotinic receptor-ionophore complex to regulate Na+ entry. SP receptors on the chromaffin cells show similar structural requirements to SP receptors in other SP responsive tissues. Binding studies on isolated chromaffin cell membranes with [4-3H-Phe]SP have shown specific binding in the nM range. In addition, at high concentrations of ACh, SP protects against nicotinic receptor desensitization. Since SP is contained in the splanchnic nerve terminals that innervate the medulla, the demonstration of SP action and SP receptors on the chromaffin cells suggests a physiological role for SP in the regulation of secretion from the adrenal medulla. Somatostatin (SS) and a number of SS analogues also inhibit release, but are approximately 15-fold less potent than SP. Leu- and Met-enkephalin, which are co-stored with adrenaline in the bovine adrenal medullary cells produce a non-specific inhibition of the nicotine-evoked release of CA, but enhance the basal release of endogenous catecholamines by a mechanism that is Ca2+-dependent, stereospecific and reversible by naloxone and naltrexone. The implication of these peptide-amine interactions for physiological processes regulating homeostasis in the adrenal are discussed.

Acetylcholinesterase↗
PubMed1983

Nicotine stimulates secretion of both catecholamines and acetylcholinesterase from cultured adrenal chromaffin cells.

There is conflicting evidence from studies on sympathetic ganglia and the adrenal medulla concerning the morphological and biochemical localization and physiological role(s) of the enzyme acetylcholinesterase (AChE). Furthermore, the origin of the AChE released from the adrenal medulla (whether from chromaffin cells or splanchnic nerve, or both) has not been firmly established. We have examined the efficacy of cholinergic agonists to release endogenous AChE and catecholamines (CA) from monolayer cultures of purified bovine adrenal chromaffin cells. The nicotinic agonist (nicotine), but not the muscarinic agonist (methacholine), released both AChE and CA from the adrenal chromaffin cells. The concomitant release of CA and AChE evoked by nicotine was Ca++ dependent with a correlation coefficient r = 0.82 (p less than 0.001). The results show that adrenal chromaffin cells in vitro, a system free of splanchnic nerve elements, can still release AChE. The finding that concomitant release of AChE and catecholamines occurs on exposure of the cells to nicotinic agonists suggests that released AChE may have a physiological role at neuroeffector junctions.

Acetylcholinesterase↗
PubMed1982

Characterization of substance P and somatostatin receptors on adrenal chromaffin cells using structural analogues.

Substance P (SP) and somatostatin (SRIF) are known to inhibit the nicotine-induced release of catecholamines (CAs) from isolated adrenal chromaffin cells in culture22,24. In order to characterize the receptors mediating this action, we have tested several SP and SRIF analogues for their effects on release of [3H]L-norepinephrine ([3H]NE) from chromaffin cell cultures. SP-free acid and a series of 11 SP analogues, in which each amino acid of SP is replaced in turn by L-alanine, all inhibited the nicotine-induced release of [3H]NE from these cultures. The rank order of potency of the analogues for this action was similar to their reported order of potency in other SP-responsive tissues. The least potent were SP-free acid, [Ala7]-SP, [Ala10]-SP, [Ala8]-SP and [Ala11]-SP, while the potencies of the Ala 1 to 6 analogues and [Ala9]-SP were closer to that of SP. [Leu7]- and [Leu7,8]-SP had potencies similar to that of SP. SP itself had no effect on basal [3H]NE release and only the highest concentrations of some of the analogues tested had an effect (enhancement) on basal [3H]NE release. The results suggest that adrenal chromaffin cells possess a specific SP receptor mediating inhibition of agonist-induced CA release and that the binding site of this receptor shares similar structural requirements with the binding site of the SP receptor on other tissues. Several SRIF analogues, which have been previously shown to be more potent than native SRIF at selective SRIF receptors2,3,31, 35, were compared to SRIF for effects on [3H]NE release from chromaffin cell cultures. These analogues were found to be active but less potent than SRIF in inhibiting nicotine-induced [3H]NE release from these cultures, suggesting that the site mediating this action differs in its structural requirements from the SRIF receptor found in some other tissues.

Adrenal Medulla↗
PubMed1982

Effects of pro-opiomelanocortin fragments on release of catecholamines from adrenal chromaffin cells.

The effects of pro-opiomelanocortin (POMC) peptide fragments on the basal and agonist-induced release of catecholamines (CAs) from monolayer cultures of purified bovine adrenal chromaffin cells were tested. None of the 5 peptides tested, i.e. beta-MSH, ACTH1-39, gamma-MSH1-13, gamma 3-MSH and N-terminal POMC fragment, had any effect on basal CA release. However, beta-MSH (10(-5) M), gamma-MSH1-13 (10(-6)-10(-5) M), gamma 3-MSH (10(-5) M) and N-terminal POMC fragment (10(-5) M) inhibited the nicotine-induced release of CAs from the chromaffin cells. The possible physiological significance of this inhibitory neuromodulation is discussed.

Adrenal Medulla↗