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B Collier

Publications and source records attributed to B Collier.

At least 91 records · Page 5Linked to original sources
PubMed1987

Effect of endogenous opioid peptides on acetylcholine release from the cat superior cervical ganglion: selective effect of a heptapeptide.

The present experiments show the presence of both metenkephalin-like and met-enkephalin-Arg6-Phe7-like immunoreactivity in the superior cervical ganglion of the cat; this was determined by radioimmunoassay after high-pressure liquid chromatography separation of tissue extracts. There was measurable efflux of both peptides, as determined by radioimmunoassay of ganglionic perfusates; this measure was increased by thiorphan, an enkephalinase inhibitor. The effect of the 2 peptides on ACh release was determined: The stable analog of methionine-enkephalin, D-Ala2-methionine-enkephalinamide, did not affect ACh release from the ganglion; in contrast, methionine-enkephalin-Arg6-Phe7 significantly depressed evoked ACh release. The effect of met-enkephalin-Arg6-Phe7 to decrease ACh release was antagonized, although only partially, by the opioid antagonist naloxone. Thus, it appears that methionine-enkephalin-Arg6-Phe7 alters ACh release from the superior cervical ganglion by acting, at least in part, on a presynaptic opioid receptor. The results suggest that in the cat superior cervical ganglion, the heptapeptide enkephalin might have a significant role in the regulation of synaptic transmission, which is unrelated to its potential function as a precursor for methionine-enkephalin.

Acetylcholine↗
PubMed1986

Evidence that endogenous catecholamines can regulate acetylcholine release in a sympathetic ganglion.

The objective of this study was to test whether activation of alpha-adrenoceptors by endogenously released catecholamines alters the release of acetylcholine (ACh) from the cat superior cervical ganglion. The alpha-adrenoceptor agonists noradrenaline and clonidine depressed evoked ACh release; this effect was concentration-dependent; it was apparent during preganglionic stimulation at 20 Hz, but not so at lower frequencies of stimulation. The inhibitory effect of noradrenaline on evoked ACh release was reversed by yohimbine, by phentolamine and, to a lesser extent, by prazosin. Thus, exogenous amines can depress evoked ACh release by an action on presynaptic alpha-adrenoceptors. To determine if activation of these receptors by endogenous amines inhibits ACh release, we tested whether the alpha-adrenoreceptor antagonists enhance ACh release. Yohimbine and phentolamine increased evoked ACh release during preganglionic stimulation at 20 Hz, but not during stimulation at 5 Hz, suggesting that endogenous, like exogenous, amine can depress evoked ACh release from preganglionic nerve terminals.

Acetylcholine↗
PubMed1986

Posterior vitreous fluorophotometry in normal subjects.

We performed vitreous fluorophotometry using the Fluorotron Master and intravenous fluorescein injection of 14 mg/kg of body weight in 22 normal subjects. Various methods of analysis were used to evaluate vitreous fluorescein concentration at 3 mm from the retina as well as averaged over the posterior 6 mm. The various methods of calculation yielded mean (+/- SD) postinjection values ranging from 1.7 +/- 1.4 to 4.3 +/- 2.3 ng/mL at 30 minutes and from 7.1 +/- 2.4 to 10.8 +/- 2.7 ng/mL at 60 minutes. The permeability index determined 60 minutes after injection ranged from 0.92 +/- 0.40 X 10(-7) to 1.19 +/- 0.30 X 10(-7) cm/s, according to the protocol used. Replicate pairs of measurements in six subjects demonstrated that the procedure was reproducible to within 17% to 49%, depending on the analysis. The results suggest that if the current methods of data analysis are used, fluorescein leakage might be considered abnormally high if at 60 minutes the 3-mm posterior vitreous fluorescein concentration corrected for background fluorescence exceeds 14.3 ng/mL and/or the permeability index exceeds 2 X 10(-7) cm/s.

Adult↗
PubMed1986

Effect of 2-(4-phenylpiperidino)cyclohexanol on acetylcholine release and subcellular distribution in rat striatal slices.

These experiments measured the effect of 2-(4-phenylpiperidino)cyclohexanol (AH5183) on the release of acetylcholine (ACh) and its subcellular distribution in slices of rat striatum incubated in vitro. The AH5183, a drug that blocks the uptake of ACh by isolated synaptic vesicles, reduced the release of ACh from slices stimulated to release transmitter in response to K+ depolarization. Tissue stimulated in the presence of AH5183 contained more ACh in a nerve terminal cytoplasmic fraction than did tissue stimulated in the drug's absence, but stimulation in AH5183's presence reduced the amount of ACh measured in fractions containing synaptic vesicles. The depletion of ACh caused by stimulating tissue in the presence of AH5183 was more evident in the fraction of nerve terminal ACh occluded within synaptic vesicles as isolated by gradient centrifugation (fraction D) than it was in other nerve terminal occluded stores. It is concluded that the synaptic vesicles isolated as fraction D under the present experimental conditions likely contain releasable transmitter. The AH5183 also depressed the spontaneous release of ACh from incubated slices of striatum and this effect was evident in the presence or the absence of medium Ca2+. It is suggested that this effect might indicate that the process of spontaneous ACh release measured neurochemically results, in part, from an AH5183-sensitive carrier-mediated process.

Acetylcholine↗
PubMed1986

Acetylcholine synthesis and release by a sympathetic ganglion in the presence of 2-(4-phenylpiperidino) cyclohexanol (AH5183).

These experiments measured the release and the synthesis of acetylcholine (ACh) by cat sympathetic ganglia in the presence of 2-(4-phenylpiperidino) cyclohexanol (AH5183), an agent that blocks the uptake of ACh into synaptic vesicles. Evoked transmitter release during short periods of preganglionic nerve stimulation was not affected by AH5183, but release during prolonged stimulation was not maintained in the drug's presence, whereas it was in the drug's absence. The amount of ACh releasable by nerve impulses in the presence of AH5183 was 194 +/- 10 pmol, which represented 14 +/- 1% of the tissue ACh store. The effect of AH5183 on ACh release was not well antagonized by 4-aminopyridine (4-AP), and not associated with inhibition of stimulation-induced calcium accumulation by nerve terminals. It is concluded that AH5183 blocks ACh release indirectly, and that the proportion of stored ACh releasable in the compound's presence represents transmitter in synaptic vesicles available to the release mechanism. The synthesis of ACh during 30 min preganglionic stimulation in the presence of AH5183 was 2,448 +/- 51 pmol and in its absence it was 2,547 +/- 273 pmol. Thus, as the drug decreased ACh release it increased tissue content. The increase in tissue content of ACh in the presence of AH5183 was not evident in resting ganglia; it was evident in stimulated ganglia whether or not tissue cholinesterase was inhibited; it was increased by 4-AP and reduced by divalent cation changes expected to decrease calcium influx during nerve terminal depolarization.(ABSTRACT TRUNCATED AT 250 WORDS)

4-Aminopyridine↗
PubMed1986

Choline analogues: their use in studies of acetylcholine synthesis, storage, and release.

The objective of this article is to illustrate how choline analogues might provide insight into mechanisms that regulate the synthesis, storage, and release of acetylcholine (ACh). Studies with false neurotransmitters provide information about the origin of releasable transmitter. Thus, false esters that distribute like ACh to vesicle-bound stores are as releasable as is ACh, but esters that poorly localize to synaptic vesicles are poorly releasable. Studies of choline analogue uptake provide information about the structural specificity of that transport process and, also, show that choline uptake is regulated in response to activity. Thus, stimuli that normally release transmitter increase the rate of choline transport, presumably to provide more precursor for ACh synthesis. However, the relationship between precursor delivery and product formed can be dissociated, suggesting that some factor in addition to choline delivery is involved in ACh synthesis regulation. Studies with a compound (AH5183), which inhibits ACh uptake by synaptic vesicles, provide information about the relationship of ACh stores and releasable transmitter. In the presence of AH5183 some 15% of nerve terminal ACh is released in response to nerve impulses, suggesting the existence of a small population of vesicles that contain readily releasable ACh. In presence of AH5183, ACh synthesis is activated even when ACh release is depressed, showing that transmitter synthesis can be regulated by some factor other than nerve terminal ACh levels.

Acetylation↗
PubMed1986

Acetylcholine release from canine isolated airway is not modulated by norepinephrine.

We measured acetylcholine (ACh) release from canine isolated tracheal smooth muscle (TSM) and bronchial spirals using a radioenzymic assay technique. Tissue was incubated in physiological salt solution containing physostigmine (3.10(-5) M), atropine (10(-7) M), and choline (5.10(-6) M), and bath fluid was collected every 15 min for assay. There was a resting release of ACh of 209 +/- 44 pmol/g tissue (mean +/- SE) from 53 to 77 specimens of TSM. Electrical field stimulation (ES) increased ACh release, which was blocked by tetrodotoxin (10(-6) g/ml), confirming the neural origin of ACh. The ACh output during ES (2-ms pulses) at 10 Hz increased linearly from 188 +/- 50 pmol/g tissue (mean +/- SE) for a 1-min volley, to 323 +/- 57 for three volleys, and 544 +/- 128 for five volleys. The ACh output/pulse was constant during ES at 20, 15, 10, and 5 Hz, but it was significantly higher at 2 than at 5 Hz (P less than 0.005). Incubation of TSM with norepinephrine (NE, 10(-5) M) did not affect ACh output either at 2 or 10 Hz. Likewise, ACh output from bronchial spirals during ES and 2 Hz was unaffected by NE. In contrast, NE treatment of isolated guinea pig ileum reduced the ACh released by ES at 2 Hz to 40 +/- 7% (P less than 0.001) of the control ACh output. It is concluded that evoked release of ACh (output/pulse) from cholinergic nerves in canine airway is frequency dependent, as in guinea pig ileum, but that, unlike guinea pig ileum, NE does not modulate its release.

Acetylcholine↗
PubMed1986

The developmental features of marrow stroma in ectopic bone marrow implants.

Implantation of bits of marrow in ectopic sites is followed by reorganization of tissue and the formation of a hemopoietic nodule surrounded by a shell of bone. This regenerative process is reminiscent of marrow ontogeny and the model can serve to study marrow ontogeny in a relatively short period of time. Early events during this regeneration were studied by scanning (SEM) and transmission electron microscopy (TEM). Within 24 hours the implant elicited an angiogenic reaction and new vessels penetrated the implant. Intense circulation, thus established, divested the implant from hemopoietic cells, leaving the stroma behind. Stromal cells proliferated and the impetus for this proliferation appeared to result from an impulse caused by the presence of bony fragments outside and within the stromal cells. Previous studies of this model have not appreciated the presence of non-viable bone in the implant, although the fact that non-viable bone can trigger osteogenesis and new bone marrow formation is well-known. This experimental model lends itself to the study of the interrelationship of hemopoietic cells and their supporting stroma as well as the interrelationship of bone and hemopoiesis.

Animals↗
PubMed1985

Accumulation, acetylation, and releasability of diethylhomocholine from a sympathetic ganglion.

Superior cervical ganglia of the cat perfused with [14C]diethylhomocholine [( 14C]DEHCh) synthesized acetyldiethylhomocholine (ADEHCh), but rather little of this ester was released by subsequent preganglionic nerve stimulation. Stimulation evoked the release of an appreciable amount of unchanged DEHCh when ganglia had been exposed to the analogue in the absence of choline (Ch), but did not do so when exposed to both Ch and DEHCh. The release of DEHCh was Ca2+ dependent, and was not the result of the release and subsequent hydrolysis of ADEHCh. This is the first clear demonstration of the release of an unacetylated compound from mammalian tissue; therefore, the characteristics of the transmitter release mechanism are further defined. The effect of preganglionic nerve stimulation on the uptake and acetylation of DEHCh was also measured. Stimulated ganglia accumulated approximately 4 times more labeled analogue and synthesized 7.5 times more ADEHCh than did rested ganglia. Stimulated ganglia perfused with 2-(4-phenylpiperidino)cyclohexanol, a compound considered to inhibit acetylcholine (ACh) release by inhibiting its transport into synaptic vesicles, accumulated 3.4 times as much and acetylated 6 times as much DEHCh as did rested ganglia. When the concentration of Mg2+ in the perfusion medium was increased to block ACh release, accumulation of the labelled analogue was enhanced by stimulation, but its acetylation was increased much less than during perfusion with normal medium. It is concluded that the synthesis of ADEHCh is subject to the same regulation as is ACh synthesis and that the activation of ester synthesis during activity can be dissociated from ester release.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylation↗
PubMed1985

Hemopoiesis in cellulose ester membrane: characterization of the epilayer responsible for recognition and lodgement of hemopoietic cells.

Following intraperitoneal (i.p.) insertion of cellulose ester membranes in the mouse, the membranes develop a cellular coat that after sublethal irradiation and i.p. infusion of marrow cells supports the growth of hemopoietic colonies. The uppermost layer of this cellular coat (the epilayer) becomes extremely attenuated and develops numerous microvilli that interact with infused cells to trap and lodge them. This phenomenon is analogous to specific lodging of hemopoietic stem cells in the marrow after bone marrow transplantation. The mechanism of this interaction is not clear, but attention has recently focused on specific interaction of sugar residues of membrane glycoproteins. We have characterized the free surface of this epilayer with various lectins, glycosylated ferritins, and antifactor VIII antibody. There is strong binding of the three lectins Ricinis communis agglutinin (RCA II), phytohemagglutinin (PHA), and wheat germ agglutinin (WGA), but not Ulex europeaus agglutinin, concanavalin A, the tested glycosylated ferritins, or antifactor VIII antibody. The highest density of binding is on microvilli. Since marrow sinus endothelium also strongly binds RCA II, PHA, and WGA, it is possible that the sugar residues of membrane glycoproteins specifically binding these lectins are responsible for cellular and molecular recognition and transport across the epilayer.

Animals↗
PubMed1984

The release of acetylcholine and of catecholamine from the cat's adrenal gland.

The cat's adrenal gland was perfused in situ with Krebs solution containing eserine; the amount of acetylcholine and of catecholamine released was measured. Splanchnic nerve stimulation (5 Hz for 2 min) increased the release of acetylcholine and catecholamine; the molar ratio of evoked release of catecholamine to acetylcholine was 122 +/- 8. It is suggested that this amplification is achieved because a chromaffin cell granule contains more mediator than does the acetylcholine quantum that releases it. The release per impulse of catecholamine during splanchnic nerve stimulation at 30 Hz was less than that released by stimulation at 1 or 5 Hz. This depression is attributed to a presynaptic failure, because the release of acetylcholine was similarly frequency dependent. The release of catecholamine was linearly related to the release of acetylcholine over the range tested, indicating that the input-output relationship at the splanchnic-adrenal medullary junction is linear. During continuous stimulation of the splanchnic nerve (5 Hz), catecholamine release declined to a level that was 32 +/- 2% of the initial output. This fatigue is attributed primarily to a postsynaptic depression, because the release of acetylcholine was maintained at 71 +/- 6% of its initial level. The presence of eserine in the perfusate was necessary for the release of acetylcholine to be detected, but in the presence of eserine catecholamine release was 90 +/- 10% that in the drug's absence. It is concluded that released acetylcholine is hydrolysed at some distance from its site of release and action. Glands perfused with raised K+ released acetylcholine and catecholamine.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1984

Uptake, metabolism, and releasability of ethyl analogues of homocholine by rat brain.

Ethyl analogues of homocholine were synthesized and used to describe further the specificities of the processes involved in choline uptake and acetylation and acetylcholine storage and release. Monoethylhomocholine, diethylhomocholine, and triethylhomocholine decreased the transport of choline into rat brain synaptosomes. The mono- and diethyl compounds were taken up into synaptosomes with similar affinity for the transport system as choline (5.8, 8.5, and 5.5 microM, respectively) but at a somewhat slower rate (11.3, 8.5, and 37.3 nmol/g original tissue/h, respectively); the triethyl analogue was not transported at the concentrations tested, which further defines the structural specificity of the transport system. L-Carnitine did not affect the transport of the analogues. The in situ acetylation of mono- and diethylhomocholine by slices of rat cerebral cortex was measurable, but the in vitro acetylation by choline acetyltransferase solubilized from rat forebrain was not. Acetylation of the diethyl analogue by slices of cerebellar cortex was less than 20% of that by slices of cerebral cortex. Subcellular fractionation of cerebral slices showed that acetyldiethylhomocholine localized preferentially to the cytosolic rather than vesicular stores, indicating specificity of the mechanism responsible for the incorporation of acetylated product into the vesicles. The release of acetyldiethylhomocholine and of acetylcholine was tested from sliced brain that had been incubated with the precursors. Both esters were released spontaneously but stimulation with increased K+ concentration enhanced the release of acetylcholine without changing the release of acetyldiethylhomocholine, suggesting that evoked transmitter release occurred from a vesicular store.

Acetylation↗
PubMed1984

Effect of chemical destruction of adrenergic neurones on some cholinergic mechanisms in adult rat sympathetic ganglia.

Rats were treated for 2-6 weeks with guanethidine after which their superior cervical ganglia were removed. Ganglionic tyrosine hydroxylase and alpha-bungarotoxin binding sites were reduced by the guanethidine treatment indicating adrenergic cell body destruction. Choline acetyltransferase activity and acetylcholine content of ganglia were not clearly changed by the guanethidine treatment, indicating that the drug does not destroy presynaptic terminals and that these presynaptic indicators do not adapt markedly to postsynaptic loss. The cholinesterase in the ganglia was reduced by guanethidine treatment, but such ganglia retained their ability to accumulate surplus acetylcholine when they were incubated with physostigmine. This is interpreted as indicating surplus acetylcholine accumulation is a presynaptic phenomenon. Choline uptake by resting ganglia was not reduced as a result of guanethidine treatment nor was it affected by preganglionic denervation. This is interpreted as indicating that during rest, choline uptake is into supporting cells or intraganglionic cells rather than cholinergic nerve terminals or adrenergic cell bodies.

Acetylcholine↗
PubMed1983

Hemicholinium impairs septo-hippocampal facilitatory action.

Intraventricular injections of hemicholinium-3 led to a sharp reduction in hippocampal acetylcholine content (by 79% on the average). This was associated with the following changes in population spikes evoked in area CA1 by commissural stimulation: (1) a tendency to progressive increase, over 1-2 hours; (2) in a few cases (3 out of 12), a striking depression of the normal strong facilitation produced by brief tetanic stimulation of the medial septum (typically 10 pulses at 50-100 Hz); (3) a much more consistent tendency towards fading of the septal facilitatory effect during repeated applications of such brief septal tetani (in 10 cases out of 12); as well as, (4) diminished facilitation by sustained, lower frequency septal tetanic stimulation (20-50 Hz). The reduced efficiency of septal action--especially during repetitive stimulation--was not accompanied by a consistent reduction of the facilitation produced by local applications of acetylcholine; it is, therefore, best explained by the diminished availability of acetylcholine, and so provides further evidence that septo-hippocampal facilitation is mediated by a cholinergic mechanism.

Acetylcholine↗
PubMed1983

Increased acetylcholine synthesis and release following presynaptic activity in a sympathetic ganglion.

The acetylcholine (ACh) content of sympathetic ganglia increases above its normal level following a period of preganglionic nerve stimulation. In the present experiments, this extra ACh that accumulates following activity was labeled radioactively from [3H]choline and its specific activity was compared with that of ACh subsequently released during preganglionic nerve stimulation. The specific activity released ACh was similar to that of the total tissue ACh, suggesting that the extra ACh mixes fully with endogenous stores. The present experiments also show that transmitter release during neuronal stimulation is necessary for the poststimulation increase in transmitter store, However, the increase was not evident when transmitter release was induced by K+. It is concluded that both transmitter release and impulse invasion of the nerve terminals are necessary for the adaptive phenomenon to manifest itself. The role of choline delivery and choline acetyltransferase activity in generating the poststimulation increase in transmitter store was tested. When choline transport activity measured as choline analogue (homocholine) accumulation increased. ACh synthesis was increased and when transport activity was not increased, neither was ACh synthesis. There was no poststimulation increase in measured choline acetyltransferase activity.

Acetyl Coenzyme A↗
PubMed1983

Effects of 4-aminopyridine on the cat superior cervical ganglion.

These experiments tested the effect of 4-aminopyridine (4-AP) on acetylcholine (ACh) release, 45Ca++ accumulation and transmission in the perfused superior cervical ganglion of the cat. The 4-AP increased the amount of ACh released during preganglionic nerve stimulation, but it did not alter spontaneous ACh release. The 4-AP-induced increase of ACh release was compensated for by increased ACh synthesis because stimulation in the presence of the drug did not deplete tissue ACh content. When ACh release was suppressed by Mg++ or by low Ca++, 4-AP restored release to normal, but it did not do so when Ca++ was absent. This is interpreted as consistent with the idea that 4-AP increases Ca++ influx into nerve terminals and this was supported by measures of 45Ca++ accumulation by ganglia. Thus, preganglionic nerve stimulation increased 45Ca++ accumulation by ganglia and 4-AP increased this measure; Mg++ decreased the stimulation-induced change in 45Ca++ accumulation and 4-AP reversed this effect of Mg++. Depression of ganglionic transmission caused by Mg++ was readily antagonized by 4-AP, but the compound did not as readily augment transmission depressed by tubocurarine or by trimethaphan.

4-Aminopyridine↗
PubMed1982

Synthesis of acetylcholine from acetate in a sympathetic ganglion.

The present experiments tested whether acetate plays a role in the provision of acetyl-CoA for acetylcholine synthesis in the cat's superior cervical ganglion. Labeled acetylcholine was identified in extracts of ganglia that had been perfused for 20 min with Krebs solution containing choline (10(-5) M) and [3H], [1-14C], or [2-14C]acetate (10(-3) M); perfusion for 60 min or with [3H]acetate (10(-2) M) increased the labeling. The acetylcholine synthesized from acetate was available for release by a Ca2+-dependent mechanism during subsequent periods of preganglionic nerve stimulation. When ganglia were stimulated via their preganglionic nerves or by exposure to 46 mM K+, the labeling of acetylcholine from [3H]acetate was reduced when compared with resting ganglia. The reduced synthesis of acetylcholine from acetate during stimulation was not due to acetate recapture, shunting of acetate into lipid synthesis, or the transmitter release process itself. In ganglia perfused with [2-14C]glucose, the amount of labeled acetylcholine formed was clearly enhanced during stimulation. An increase in acetylcholine labeling from [3H]acetate was shown during a 15-min resting period following a 60-min period of preganglionic nerve stimulation (20 Hz). It is concluded that acetate is not the main physiological acetyl precursor for acetylcholine synthesis in this sympathetic ganglion, and that during preganglionic nerve stimulation there is enhanced delivery of acetyl-CoA to choline acetyltransferase from a source other than acetate.

Acetates↗