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Biomedical subjects

B G Livett

Publications and source records attributed to B G Livett.

At least 91 records · Page 5Linked to original sources
PubMed1982

Evidence that inhibition of nicotine-mediated catecholamine secretion from adrenal chromaffin cells by enkephalin, beta-endorphin, dynorphin (1-13), and opiates is not mediated via specific opiate receptors.

The opioid peptides Met- and Leu-enkephalin, dynorphin (1-13), and beta-endorphin and the narcotic analgesics, morphine, levorphanol, and dextrorphan all produced a dose-dependent inhibition of nicotine (5 x 10(-6) M)-mediated release of [3H]norepinephrine ([3H]NE) from bovine adrenal chromaffin cells in culture. None of these agents affected [3H]NE release induced by high K+ (56 mM). Although the above results suggest that the opioid peptides and narcotic analgesics inhibit catecholamine release from adrenal chromaffin cells in culture, we suggest that these effects are not mediated by specific opiate binding sites, since (1) the inhibition was only produced with high concentrations of the agents--the threshold concentrations were 10(-7) to 10(-5)M and higher; (2) the inhibition produced by the narcotic analgesics did not display stereospecificity, because the d-isomer, dextrorphan, was slightly more active than the l-isomer, levorphanol; (3) the narcotic antagonists naloxone, naltrexone, and levallorphan did not reverse the inhibition produced by either the narcotic analgesics (e.g., morphine) or the opioid peptides (e.g., dynorphin). These three antagonists themselves inhibited the nicotine-mediated release of [3H]NE from the adrenal chromaffin cells in culture. Finally (4), the I2-Tyr1 substituted analogues of beta-endorphin and dynorphin that are biologically less active than the parent compounds produced an inhibition of the nicotine-mediated [3H]NE release similar to that of their parent compounds. These results do not support the idea that high-affinity stereospecific opiate binding sites are involved in the inhibitory modulation of nicotinic evoked catecholamine release from bovine adrenal chromaffin cells in culture.

Adrenal Glands↗
PubMed1981

Co-release of enkephalin and catecholamines from cultured adrenal chromaffin cells.

The opioid peptides Leu-enkephalin and Met-enkephalin are stored intraneuronally in the brain where they are thought to act as neurotransmitters and/or neuromodulators. Evidence for their release from nerve terminals has come from biochemical and pharmacological studies in vitro with brain tissue slices and synaptosomes. Enkephalins also exist in the peripheral nervous system in nerve cell bodies and axon terminals in the gastrointestinal tract, sympathetic ganglia and adrenal gland. In the adrenal gland, high levels of enkephalins are present both in axon terminals of the splanchnic nerve and in the adrenal medullary chromaffin cells where they are stored together with the catecholamines in the chromaffin granules. Stimulation of the adrenal gland in vivo or the perfused gland in vitro causes release of catecholamines and enkephalins into the adrenal vein. However, it is not clear whether the origin of the released enkephalins is the adrenal medullary chromaffin cells or the enkephalin-containing splanchnic nerve terminals that innervate the medulla. We now show that enkephalin and catecholamines are released together from primary cultures of bovine adrenal medullary chromaffin cells by nicotine in a Ca2+-dependent manner.

Adrenal Medulla↗
PubMed1980

Production and release of acetylcholinesterase by a primary cell culture of bovine adrenal medullary chromaffin cells.

Bovine adrenal chromaffin cells were isolated and maintained as primary culture monolayers. Total acetylcholinesterase (AChE) activity in the cells increased during the culture period, and AChE activity appeared in the culture medium. We have examined the role of the AChE synthesized by the cells on ACh-evoked release of catecholamine from the cells. A progressive decrease in the efficacy of ACh (5 X 10(-5) M) to evoke release of [3H]norepinephrine from day 3-15 cultures suggests that exogenously applied ACh is hydrolyzed by the nascent AChE synthesized by the cells. These findings provide evidence that chromaffin cells produce AChE and release it into their immediate environment.

Acetylcholine↗
PubMed1979

Pharmacological characterization of adrenal paraneurons: substance P and somatostatin as inhibitory modulators of the nicotinic response.

A pharmacological study was made of the effects of various muscarinic and nicotinic agonists and their antagonists on the release of [3H]noradrenaline ([3H]NA) from cultures of isolated bovine adrenal medullary cells. A study was also made of the effects of substance P and somatostatin on the release of [3H]NA evoked by nicotinic agonists. By 2 days in culture these adrenal 'paraneurons' had developed long varicose processes with growth cones and generally resembled noradrenergic neurons in culture. In the present study, adrenal paraneurons were incubated with [3H]NA which was taken up and stored in reserpine-sensitive sites. Exposure of the cultures to acetylcholine (ACh) resulted in release of [3H]NA into the external medium. High concentrations of K+ (56 mM) also evoked release of [3H]NA. The release of [3H]NA induced by ACh or K+ (56 mM) was Ca2+-dependent. Pharmacological studies with nicotinic (ACh, nicotine) and muscarinic (methacholine, pilocarpine) agonists and their antagonists (mecamylasmine, d-tubocurarine, hexamethonium; and atropine, scopolamine, respectively) showed that the adrenal paraneurons contained only nicotinic receptors. Substance P produced a dose-dependent inhibition of ACh (5 x 10(-5) M) stimulated [3H]NA release in the range of 10(-8) to 5 x 10(-5) M with an ID50 of 10(-6) M. A similar inhibition of NA release by substance P was obtained when nicotine (K X 10(-6) M) was used as the agonist, but not when K+ (50 MM) was used to depolarize the cells. Substance P (10-10) to 5 x 10(-5) M) by itself did not have a significant effect on the basal release rate of [3H]NA from these cells. Somatostatin at relatively high concentrations (10(-6)-10(-3) M; ID50 2 x 10(-5) M) inhibited the release induced by ACh, but not by K+ (56 mM). The present results provide the first direct evidence at a cellular level that substance P and somatostatin act as inhibitory modulators of the nicotinic ACh response, and support a role for these peptides as inhibitory neuromodulators at nicotinic receptor sites in the nervous system.

Acetylcholine↗
PubMed1979

Experimental autonomic neuropathy: ultrastructure and immunohistochemical study of a disorder of reflex vasomotor function.

Vasomotor function in ear vessels of rabbits was tested in animals with experimental autonomic neuropathy (EAUN), experimental allergic neuritis (EAN), and controls. Reflex vasodilatation was impaired in animals with EAUN but not in EAN or control rabbits. Ultrastructural examination of paravertebral sympathetic chain, perivascular nerves, and perivascular tissue showed similar changes in all animals differing in degree only. Occasional lymphocytes were seen in paravertebral chains of controls but large numbers of lymphocytes and macrophages occurred in EAUN and EAN. No active destruction of myelinated or unmyelinated fibers could be seen. Perivascular nerves showed similar cellular infiltrates and basophils were found in perivascular spaces of ear vessels which were most abundant in animals with EAUN. Fibre-size spectra analysis of unmyelinated fibres in the paravertebral sympathetic chain showed a shift to the left suggesting regeneration of unmyelinated axons in both EAN and EAUN. No specific immunofluorescence against neuronal components could be demonstrated in sensitized animals. It is concluded that the functional disturbance of reflex vasomotion in EAUN is not accompanied by specific ultrastructural lesions nor could neuronal components be immunologically implicated in the disorder. Regeneration of unmyelinated axons and recovery of function after some weeks together with abundance of basophils in perivascular tissue of sensitized animals, suggests that a localized inflammation may have occurred, the basophils interfacing between foreign antigens, the serum cascade systems, and other inflammatory cells. The usefulness of EAUN as a model for acute pandysautonomia in man remains to be determined.

Animals↗
PubMed1978

Functional and morphological characterization of isolated bovine adrenal medullary cells.

Single bovine adrenal medullary cells have been obtained by retrograde perfusion of adrenal medullae with a solution of 0.05% collagenase in Ca++-free Krebs Henseleit buffer. Chromaffin cells were obtained in high yield (5 X 10(6) cells/g medulla), and more than 95% of these were viable as shown by exclusion of trypan blue. The isolated cells were capable of respiring at a linear rate for a minimum of 120 min. Ultrastructural examination revealed that the cells were morphologically intact, and two distinct types of adrenal medullary cells were identified, on the basis of the morphology of their electron-dense vesicles, as (a) adrenaline-containing and (b) noradrenaline-containing cells. Biochemical analysis showed that the cells contained catecholamines and dopamine-beta-hydroxylase (DBH). The cells released catecholamines and DBH in response to acetylcholine (ACh), and this release was accompanied by changes in the vesicular and surface membranes observed at the ultrastructural level. The time-course of ACh-stimulated catecholamine and DBH release, and the dependence of this release on the concentration of ACh and extracellular Ca++ have been investigated. The isolated cells were pharmacologically sensitive to the action of the cholinergic blocking agents, atropine and hexamethonium.

Adrenal Medulla↗
PubMed1977

Skeletal-muscle sarcolemma from normal and dystrophic mice. Isolation, characterization and lipid composition.

1. Mouse skeletal-muscle sarcolemma was isolated, and the preparations obtained from normal mouse muscle and from muscle of mice with hereditary muscular dystrophy were characterized with respect to appearance under the optical and electron microscopes, distribution of marker enzymes, histochemical properties and biochemical composition. 2. The sarcolemmal membranes from normal and dystrophic muscle were subjected to detailed lipied analysis. Total lipid content was shown to increase in sarcolemma from dystrophic mice as a result of a large increase in neutral lipid and a smaller increase in total phospholipids. Further analysis of the neutral-lipid fraction showed that total acylglycerols increased 6-fold, non-esterified fatty acid 4-fold and cholesterol esters 2-fold, whereas the amount of free cholesterol remained unchanged in sarcolemma from dystrophic muscle. Significant increases were found in lysophosphatidylcholine, phosphatidylcholine and phosphatidylethanolamine in dystrophic-muscle sarcolemma; however, the relative composition of the phospholipid fraction remained essentially the same as in the normal case. 3. The overall result of alterations in lipid composition of the sarcolemma in mouse muscular dystrophy was an increase in neutral lipid compared with total phospholipid, and a 4-fold decrease in the relative amount of free cholesterol in the membrane. The possible impact of these changes on membrane function is discussed.

Animals↗
PubMed1977

Immunochemical comparison of synaptic plasma membrane and synaptic vesicle membrane antigens.

A synaptic vesicle fraction and a synaptic plasma membrane fraction obtained after subfractionation of synaptosomes from chick forebrain have been used to produce antisera in rabbits. Immunofluorescence histology with the two antisera revealed that they reacted strongly with synaptic terminal regions present in the chick forebrain, cerebellum and spinal cord. In addition, the synaptic plasma membrane antiserum (but not the synaptic vesicle antiserum) reacted with preterminal axons in the cerebellum and spinal cord. Comparison of the two antisera by two-dimensional immunoelectrophoresis, revealed the presence of common antigens in the synaptosomal vesicle and plasma membrane fractions. Incubation of synaptosomes in vitro with the synaptosomal vesicle antiserum and complement produced a dose-dependent inhibition of synaptosome swelling up to a maximum of 55% of that obtained with the synaptosomal plasma membrane antiserum. The results of this test are consistent with the hypothesis that some synaptosomal vesicle antigens may be present also in the synaptosomal plasma membrane and imply that they face the external surface of the synaptosomes. The fate of vesicle membrane components in synaptosomal plasma membranes is not known. The possibility is discussed that they may be recycled locally by a mechanism similar to that proposed by Heuser and Reese (1973) for re-use of synaptic vesicle membranes at the neuromuscular junction.

Animals↗