Search PubMed⌕ Search

Biomedical subjects

B Collier

Publications and source records attributed to B Collier.

At least 37 records · Page 2Linked to original sources
PubMed1994

Stereoselectivity of the inhibition of [3H]hemicholinium-3 binding to the sodium-dependent high-affinity choline transporter by the enantiomers of alpha- and beta-methylcholine.

In a previous report, we showed that the enantiomers of alpha- and beta-methylcholine inhibited choline uptake with stereoselectivity, but that their transport by the choline carrier of nerve terminals showed stereospecificity. The present experiments used the same choline analogues to determine if either of the above characteristics pertains to their ability to interact with the [3H]-hemicholinium-3 binding site present on striatal membranes and synaptosomes. [3H]Hemicholinium-3 binding to striatal membranes could be inhibited stereoselectively by the enantiomers of beta-methylcholine, but R(+)-alpha-methylcholine was little better than its enantiomer in this test. However, [3H]hemicholinium-3 binding to striatal synaptosomes was inhibited stereoselectively by the enantiomers of both alpha- and beta-methylcholine. This difference between the properties of [3H]hemicholinium-3 binding to membranes or to synaptosomes appears related to the presence of two ligand binding states. The [3H]hemicholinium-3 binding site could be shifted to a low-affinity state by ATP treatment and to a high-affinity state by EDTA washing. When the [3H]hemicholinium-3 binding site existed in its low-affinity state, binding was inhibited stereoselectively by the enantiomers of both alpha- and beta-methylcholine, but when shifted to its high-affinity state, it was inhibited stereoselectively only by the enantiomers of beta-methylcholine. We conclude that hemicholinium-3 interacts with the substrate recognition site of the high-affinity choline transporter, but that the stereoselectivity of this site changes depending on its affinity state.

Animals↗
PubMed1994

Regulation of rat brain synaptosomal [3H]hemicholinium-3 binding and [3H]choline transport sites following exposure to choline mustard aziridinium ion.

Choline uptake by cholinergic nerve terminals is increased by depolarization; the literature suggests that this results from either the appearance of occult transporters or the increased activity of existing ones. The present experiments attempt to clarify the mechanism by which choline transport is regulated by testing if the preexposure of synaptosomes to choline mustard aziridinium ion prevents the stimulation-induced appearance of hemicholinium-3 binding sites and/or choline transport activity. Choline mustard inhibited irreversibly most of the "ground-state" (basal) high-affinity choline transport but only 50% of "ground-state" hemicholinium-3 binding sites. Exposure of both striatal and hippocampal synaptosomes to the mustard, before stimulation, inhibited K(+)-stimulated increases in choline transport and of [3H]-hemicholinium-3 binding. We conclude that the mechanism by which choline transport is regulated involves the increased activity of a pool of transport sites that are occluded to hemicholinium-3 but are available to choline mustard aziridinium ion, and presumably to choline, before stimulation. However, the concentration of mustard needed to inhibit the stimulation-induced increase of [3H]-hemicholinium-3 binding and choline transport was lower for striatal synaptosomes than for hippocampal synaptosomes. In the absence of extracellular Ca2+ or presence of high Mg2+ levels, the choline mustard did not prevent the appearance of extra striatal hemicholinium-3 binding sites. Also, high Mg2+ levels removed the ability of the mustard to inhibit K(+)-stimulated increases of either [3H]-hemicholinium-3 binding or choline transport by hippocampal synaptosomes. In contrast, the preexposure of hippocampal synaptosomes to the mustard in the presence of a calcium ionophore (A23187) reduced the concentration of inhibitor needed to prevent the activation of [3H]hemicholinium-3 binding and choline uptake. Thus, we conclude that the ability of the choline mustard to alkylate the pool of choline transporters that are activated by stimulation appears dependent on the entry of extra-cellular Ca2+.

Animals↗
PubMed1994

GRBase, a new gene regulation data base available by anonymous ftp.

The Gene Regulation Database (GRBase) is a compendium of information on the structure and function of proteins involved in the control of gene expression in eukaryotes. These proteins include transcription factors, proteins involved in signal transduction, and receptors. The database can be obtained by FTP in Filemaker Pro, text, and postscript formats. The database will be expanded in the coming year to include reviews on families of proteins involved in gene regulation and to allow online searching.

Computer Communication Networks↗
PubMed1994

Pre- and postsynaptic components of nicotinic long-term potentiation in the superior cervical ganglion of the cat.

1. In anesthetized, atropine-treated cats we measured the acetylcholine (ACh) release into the venous effluent of the superior cervical ganglion (SCG) and the nictitating membrane (NM) contraction evoked by a 2-Hz, 20-s test train to the ipsilateral cervical sympathetic trunk (CST). We also measured NM contraction produced by injection of ACh (50 micrograms) or 1,1-dimethyl-4-phenylpiperazinium (DMPP, 5 micrograms) into the arterial supply of the ipsilateral SCG. 2. After a 10- to 30-s, 40-Hz conditioning train to CST these responses were all potentiated. The potentiation of the NM response evoked by the test train or by the exogenous agonists was long lasting (90% decay in 64 +/- 10 min, mean +/- SE, for the train-evoked response; 42 +/- 9 min for the response to injected ACh; 61 +/- 18 min for the response to injected DMPP), whereas the potentiation of the ACh release lasted only for 9 min. 3. On the assumption that ACh and DMPP injected into the ganglionic arterial supply acted postsynaptically, these data suggest that the main mechanism of the long-term potentiation (LTP) of nicotinic transmission in SCG is an increase in postsynaptic responsiveness. Because the response to KCl (250 micrograms) injected into the ganglionic arterial supply was not potentiated after the conditioning train, a posttrain increase in excitability of the postsynaptic membrane is not likely to be the cause of the postsynaptic increased responsiveness to ACh and DMPP.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1994

The role of endogenous adenosine in a poststimulation increase in the acetylcholine content of a sympathetic ganglion.

Previous experiments showed that exposure of sympathetic ganglia to exogenous adenosine increased acetylcholine (ACh) content and its subsequent release. This effect was not mediated through extracellular adenosine receptors, but at an intracellular site following its uptake through nitrobenzylthioinosine (NBTI)-resistant nucleoside transporters. We postulated that endogenous adenosine may play a role in modulating synaptic transmission in the superior cervical ganglion. The present study tested whether adenosine is involved in the activation of ACh synthesis that occurs during a rest period following prolonged presynaptic tetanic activity. Conditioning of ganglia with high-frequency stimulation (15 Hz) for 45 min followed by a 15 min rest increased their ACh content by 45%. The appearance of this "rebound ACh" showed sensitivity to nucleoside transport inhibitors; it was prevented by dipyridamole, but not by NBTI or meclonazepam, and it was reduced in the presence of RO 11-3624, suggesting an involvement of NBTI-resistant transporters. The effect of dipyridamole was specific for the synthesis of rebound ACh in that it did not inhibit ACh release or ACh synthesis during stimulation. The inhibitory action of dipyridamole on the synthesis of rebound ACh was not evident if it was present only during the tetanic stimulation but it was if dipyridamole was present during the rest period following it, suggesting that adenosine's presence after tetanic stimulation is of importance. This conclusion was strengthened by experiments showing that the presence of cyclopentyltheophylline, an antagonist at inhibitory adenosine receptors, increased ACh output evoked by test stimulation immediately following tetanic activity, as if endogenous adenosine was available at that time to activate the adenosine receptors that inhibit transmitter release. ACh release from conditioned ganglia was 44% greater than that from the controls. However, the rebound ACh was not mobilized in the presence of 2-(4-phenylpiperidino)cyclohexanol (vesamicol), a vesicular ACh transporter inhibitor. These results suggest that endogenous adenosine released after tetanic stimulation activates ACh synthesis, which results in an increase of ganglionic ACh that is available for subsequent mobilization and release.

Acetylcholine↗
PubMed1993

Ultrastructural localization of neurotensin immunoreactivity in the stellate ganglion of the cat.

The morphological features and cellular relationships of neurotensin-containing axon terminals were studied at light and electron microscopic levels in the cat stellate ganglion using peroxidase and immunogold immunocytochemistry. By light microscopy, neurotensin immunoreactivity was detected within thin varicose fibres distributed throughout the ganglion. Immunoreactive fibres were no longer apparent following chronic deafferentation of the ganglion indicating that they were of extrinsic origin. Ultrastructural analysis of peroxidase immunostained material confirmed the presence of neurotensin immunoreactivity within a subpopulation of axonal varicosities which made synaptic contacts with the dendrites of ganglion cells. Within labelled varicosities neurotensin immunoreactivity was found by both immunoperoxidase and immunogold methods to be concentrated within large dense core vesicles 80-120 nm in diameter. These large dense core vesicles were characteristically distant from the active zone, in keeping with a possible extrasynaptic release of the peptide.

Animals↗
PubMed1993

The ovine corticotropin-releasing hormone-stimulation test in type I diabetic patients and controls: suggestion of mild chronic hypercortisolism.

We examined hypothalamic-pituitary-adrenal (HPA) axis function in insulin-dependent diabetic outpatients (N = 22) and age-, sex-, and weight-matched normal controls (N = 22). The evaluation included measurements of 9:00 AM fasting plasma cortisol and cortisol-binding globulin (CBG) levels, 24-hour urinary free cortisol (UFC) excretion, and plasma corticotropin and cortisol responses to intravenously administered ovine corticotropin-releasing hormone ([CRH] 1 microgram/kg given as a bolus at 8:00 PM). Diabetic patients had significantly elevated 9:00 AM plasma cortisol levels (mean +/- SE, 300.7 +/- 99.3 v 237.3 +/- 99.3 nmol/L, P < .04), higher 24-hour UFC excretion (313.2 +/- 112.6 v 244.2 +/- 69.3 nmol/24 h, P < .02), and greater cortisol responses to CRH infusion (time-integrated values: 49,408.2 +/- 11,289.8 v 40,217.9 +/- 7,228.6 nmol/L.120 min, P < .004; peak cortisol values: 529.7 +/- 107.6 v 438.7 +/- 77.3 nmol/L, P < .002) than controls. UFC excretion values were positively correlated with both 5-year averaged hemoglobin A1c level (P = .03) and total number of insulin units administered per day (P = .03). These results suggest that insulin-dependent diabetic outpatients have mild chronic hypercortisolism, which might influence the control of the disease and play a role in the development of its chronic complications.

Adrenocortical Hyperfunction↗
PubMed1993

Dynamics of large dense-cored vesicles in synaptic boutons of the cat superior cervical ganglion.

We have shown previously that stimulation of the cat cervical sympathetic trunk for 2 h at 40 Hz depletes the large dense-cored vesicle store in synaptic boutons of the superior cervical ganglion and that post-depletion recovery of the store takes several days. In the present study, we examine the properties of the depletion and recovery mechanisms. Invaginations of the plasmalemma suggestive of the exocytosis of dense cores were seen frequently in boutons from stimulated ganglia. The depletion process is calcium dependent: in ganglia perfused with calcium-free Krebs solution no depletion was produced by 40 Hz preganglionic stimulation. The depletion process is rapid: during continuous stimulation of the cervical sympathetic trunk with 40 Hz, depletion observed by the end of 2 h was similar to depletion by the end of the initial 5 min of stimulation. The depletion process is frequently dependent: when the cervical sympathetic trunk was stimulated with a constant number of stimuli, no depletion occurred at the frequency of 2 or 10 Hz, while the frequencies of 20 and 40 Hz produced depletion, which was greater at 40 Hz. Recovery of the large dense-cored vesicle store during the initial 24 h after 10 min of 40 Hz stimulation was faster, and of approximately the same magnitude, than during the succeeding five days. Recovery of the store after stimulus-evoked depletion was prevented by application of colchicine to the cervical sympathetic trunk, which suggests dependence of recovery on fast axonal transport. Large dense-cored vesicles accumulated in the colchicine-treated segment of cervical sympathetic trunk axons. In conclusion, these observations suggest that the stimulus-evoked depletion of large dense-cored vesicle stores in synaptic boutons of the cat superior cervical ganglion is the result of calcium-dependent exocytosis of the large dense-cored vesicle core and that the post-stimulus recovery is critically dependent on microtubule-mediated axonal transport.

Animals↗
PubMed1993

Increased acetylcholine content induced by adenosine in a sympathetic ganglion and its subsequent mobilization by electrical stimulation.

The present study was initiated to examine the effects of ATP on acetylcholine (ACh) synthesis. The exposure of superior cervical ganglia to ATP increased ACh stores by 25%, but this effect was also evident with ADP, AMP, and adenosine, but not with beta gamma-methylene ATP, a nonhdydrolyzable analogue of ATP, or with inosine, the deaminated product of adenosine. Thus, we attribute the enhanced ACh content caused by ATP to the presence of adenosine derived from its hydrolysis by 5'-nucleotidase. The adenosine-induced increase of tissue ACh was not the consequence of an adenosine-induced decrease of ACh release. The extra ACh remained in the tissue for more than 15 min after the removal of adenosine, but it was not apparent when ganglia were exposed to adenosine in a Ca(2+)-free medium. Incorporation of radiolabelled choline into [3H]ACh was also enhanced in the presence of adenosine, suggesting an extracellular source of precursor. Moreover, the synthesis of radiolabelled forms of phosphorylcholine and phospholipid was not reduced in adenosine's presence, suggesting that the extra ACh was not likely derived from choline destined for phospholipid synthesis. Aminophylline did not prevent the adenosine effect to increase ACh content; this effect was blocked by dipyridamole, but not by nitrobenzylthioinosine (NBTI). In addition, two benzodiazepine stereoisomers known to inhibit stereoselectively the NBTI-resistant nucleoside transporter displayed a similar stereoselective ability to block the effect of adenosine. Together, these results argue that adenosine is transported through an NBTI-resistant nucleoside transporter to exert an effect on ACh synthesis. The extra ACh accumulated as a result of adenosine's action was releasable during subsequent preganglionic nerve stimulation, but not in the presence of vesamicol, a vesicular ACh transporter inhibitor. We conclude that the mobilization of ACh is enhanced as a result of adenosine pretreatment.

Acetylcholine↗
PubMed1993

Mobilization of a vesamicol-insensitive pool of acetylcholine from a sympathetic ganglion by ouabain.

These experiments investigate the release of transmitter from the perfused superior cervical ganglia of cats induced by ouabain in the absence or presence of 2-(4-phenylpiperidino)cyclohexanol (vesamicol), a blocker of acetylcholine (ACh) uptake. Ouabain, perfused through the ganglia, released ACh in a Ca(2+)-dependent way. Vesamicol caused some inhibition of the release of ACh by ouabain; however, under this condition, the Na+,K(+)-ATPase inhibitor released five times more transmitter than did preganglionic stimulation at 5 Hz. Also, when ganglia exposed to vesamicol were depleted of the impulse-releasable pool of ACh, subsequent perfusion with ouabain released ACh, and this included ACh newly synthesized in the presence of vesamicol; this phenomenon could be inhibited by the lack of Ca2+ and presence of EGTA, and was completely abolished by perfusion with a medium containing 18 mM Mg2+. To test whether the release of this vesamicol-insensitive Ca(2+)-dependent pool by ouabain is associated with a decrease in the number of synaptic vesicles, ganglia treated with the ATPase inhibitor after the depletion of the impulse-releasable pool of ACh were fixed for electron microscopy. In the presence of Ca2+, coincident with the release of the vesamicol-insensitive pool of ACh, nerve terminals were almost depleted of synaptic vesicles; ganglia treated similarly, but with medium containing 18 mM Mg2+ instead of Ca2+, were not depleted of synaptic vesicles. These results suggest that ouabain releases a vesamicol-insensitive pool of ACh from the sympathetic ganglion and also support the notion that this compartment is vesicular and its exocytosis depends on extracellular Ca2+. It is suggested that empty-vesicle recycling in the presence of vesamicol restricts mobilization of full vesicles to release sites.

Acetylcholine↗
PubMed1992

Epidermal growth factor affects both glia and cholinergic neurons in septal cell cultures.

The effects of epidermal growth factor on high density primary cultures of fetal (embryonic day 17) rat septal cells were examined. Under serum-free conditions, the continuous exposure of these cultures to epidermal growth factor for seven days significantly decreased choline acetyltransferase (EC 2.3.1.6) activity in a dose-dependent manner. Maximal decreases were observed from 1 to 10 ng/ml epidermal growth factor. This effect was completely abolished by the addition of anti-epidermal growth factor antibodies. The epidermal growth factor-mediated decrease in choline acetyltransferase activity was culture-time dependent, being first detectable after five days of factor application and may likely represent an inhibition of the spontaneous increase in enzyme activity that occurs with time in culture. Concomitant with changes in enzyme activity, epidermal growth factor produced a significant and proportional decrease in the number of acetylcholinesterase-positive neurons. This decrease in acetylcholinesterase-positive cells did not reflect a decrease in cholinergic cell survival as nerve growth factor could restore the number of acetylcholinesterase-positive neurons in epidermal growth factor-treated cultures to control levels. Furthermore, in these high-density cultures, epidermal growth factor did not affect general neuronal survival, while it did produce an increase in the number and intensity of glial fibrillary acidic protein-immunoreactive astroglia as well as in the number of macrophage-like cells. The proliferative response of these non-neuronal cells to epidermal growth factor, as assessed by [3H]thymidine incorporation, was evident after three days of epidermal growth factor application, persisted thereafter, and could be antagonized by the inclusion of the antimitotic 5-fluorodeoxyuridine. Furthermore, 5-fluorodeoxyuridine completely blocked the epidermal growth factor-mediated decrease in choline acetyltransferase activity. However, when epidermal growth factor was tested in pure glial cultures, it only directly induced proliferation of astrocytes. These results suggest that the proliferative response of either one or both of these glial cell types in the mixed cultures may be indirectly affecting cholinergic cell expression.

Acetylcholinesterase↗
PubMed1992

Role of electric stimulation in bladder evacuation following spinal cord transection.

Neural stimulation is potentially a valuable therapeutic tool in the treatment of neurogenic bladder with detrusor areflexia. We studied 20 dogs in different groups, up to eight months, and compared the effect of electric stimulation with intermittent catheterization, specially during spinal shock phase. We found that early stimulation hastened the return of detrusor activity. When stimulation was delayed, the bladder could still regain its activity, and when the pace-maker was turned off, the detrusor activity was gradually decreased. Neurostimulation can completely empty bladder up to eight months. The lowest A.Ch. content in detrusor muscle was found in intermittent catheterization group and in this group the detrusor strips showed marked supersensitivity to urecholine stimulation than the bladders managed by electric stimulation. Also, we found that electric stimulation reduced the complications caused by intermittent catheterization and protected kidney function.

Animals↗
PubMed1992

Mobilization of the readily releasable pool of acetylcholine from a sympathetic ganglion by tityustoxin in the presence of vesamicol.

The present experiments tested whether preganglionic stimulation and direct depolarization of nerve terminals by tityustoxin could mobilize similar or different pools of acetylcholine (ACh) from the cat superior cervical ganglia in the presence of 2-(4-phenylpiperidino)cyclohexanol (vesamicol, AH5183), an inhibitor of ACh uptake into synaptic vesicles. In the absence of vesamicol, both nerve stimulation and tityustoxin increased ACh release. In the presence of vesamicol, the release of ACh induced by tityustoxin was inhibited, and just 16% of the initial tissue content could be released, a result similar to that obtained with electrical stimulation under the same condition. When the impulse-releasable pool of ACh had been depleted, tityustoxin still could release transmitter, amounting to some 10% of the ganglion's initial content. This pool of transmitter seemed to be preformed in the synaptic vesicles, rather than synthesized in response to stimuli, as tityustoxin could not release newly synthesized [3H]ACh formed in the presence of vesamicol, and hemicholinium-3 did not prevent the toxin-induced release. In contrast to the results with tityustoxin, preganglionic stimulation could not release transmitter when impulse-releasable or toxin-releasable compartments had been depleted. Our results confirm that vesamicol inhibits the mobilization of transmitter from a reserve to a more readily releasable pool, and they also suggest that, under these experimental conditions, there might be some futile transmitter mobilization, apparently to sites other than nerve terminal active zones.

Acetylcholine↗
PubMed1992

A tutorial/essay project to expand the learning experience in undergraduate medical pharmacology.

To lessen fact overload and to enhance the teaching of pharmacologic principles in an undergraduate course we introduced a tutorial/essay project into our traditional lecture-based course. Under the guidance of a faculty tutor, each student researched an assigned topic, developed a paper incorporating the results of the literature review and his or her own attitudes and formally discussed and answered questions on the topic with the tutor. Students and tutors were enthusiastic about this learning experience especially in that it required independent student-centred activity, provided opportunity for useful student/tutor interaction and helped students to learn to integrate scientific information with their own thoughts and ideas.

Curriculum↗
PubMed1991

Color vision and age in a normal North American population.

Color vision is known to change with age. We conducted the Farnsworth-Munsell (FM) 100-Hue and the Lanthony Desaturated Panel D-15 (DD-15) tests in 115 normal North American subjects aged 5-81 years to obtain age-specific norms for these procedures. For each test, color discrimination was best between the ages of 20 and 50 years. Both increasing age and the occurrence of lens changes were significantly associated with increasing 100-Hue error scores. Age-specific norms for the 100-Hue test were comparable with those obtained by Verriest in a European population, but such norms for the DD-15 test are problematic. Our data indicate somewhat greater variability in the DD-15 than in the 100-Hue test.

Adolescent↗