Biomedical subjects

A Beaudet

Publications and source records attributed to A Beaudet.

170 records · Page 10Linked to original sources

PubMedDate not supplied

125I-FK 33-824: a selective probe for radioautographic labeling of mu opioid receptors in the brain.

The selectivity of the Met-enkephalin analog FK 33-824 (FK) for mu opioid receptors has been, over the years, a matter of controversy. We report here pharmacological and radioautographic data demonstrating that at nanomolar concentrations. 125I-FK interacts exclusively with mu sites. (1) Specific binding of 125I-FK to rat striatal membranes is totally inhibited by mu- and/or delta-preferring ligands according to monovalent, Michaelian kinetics, with a potency proportional to the affinity of competing drugs for mu receptors. (2) Unlabeled FK competes only at high concentration with the delta-selective ligand 3H-DPLPE and according to the same kinetics as the mu-selective agonist DAGO. (3) 125I-FK generates the same regional radioautographic labeling pattern as 3H-DAGO. We conclude that when used at nanomolar concentrations 125I-FK constitutes a selective probe for the radioautographic detection of mu opioid receptors at both light and electron microscopic levels.

Animals↗

PubMedDate not supplied

Anatomical substrate for neurotensin-acetylcholine interactions in the rat basal forebrain.

We have previously shown by combined radioautography and acetylcholinesterase histochemistry that the distribution of 125I-neurotensin (NT) binding sites was in register with that of cholinergic neurons in the rat nucleus basalis magnocellularis (NBM). The present study utilized three experimental approaches to elaborate on the type and cellular localization of NT binding sites in the NBM. Competition studies using levocabastine, a selective blocker of the low affinity NT binding component, revealed that most of the 125I-NT binding sites labeled in the NBM are of the levocabastine-insensitive high affinity type, known to correspond to the physiologically active receptor. Ibotenic acid-induced lesions of the NBM produced a marked reduction in both cholinesterase reactivity and cellular 125I-NT binding suggesting that most of the labeled sites are associated with the cholinergic neurons themselves rather than with an afferent input to those cells. Finally, examination of the high resolution radioautographic distribution of 125I-NT binding sites in semithin sections revealed that a proportion of 125I-NT-labeled receptors is associated with the plasma membrane of magnocellular perikarya and proximal processes, thereby providing an anatomical substrate for a local action of NT in the NBM.

Acetylcholine↗

PubMedDate not supplied

Estrogen-induced hypothalamic beta-endorphin neuron loss: a possible model of hypothalamic aging.

Over the course of normal aging, all female mammals with regular cycles display an irreversible arrest of cyclicity at mid-life. Males, in contrast, exhibit gametogenesis until death. Although it is widely accepted that exposure to estradiol throughout life contributes to reproductive aging, a unified hypothesis of the role of estradiol in reproductive senescence has yet to emerge. Recent evidence derived from a rodent model of chronic estradiol-mediated accelerated reproductive senescence now suggests such a hypothesis. It has been shown that chronic estradiol exposure results in the destruction of greater than 60% of all beta-endorphin neurons in the arcuate nucleus while leaving other neuronal populations spared. This loss of opioid neurons is prevented by treatment with antioxidants indicating that it results from estradiol-induced formation of free radicals. Furthermore, we have shown that this beta-endorphin cell loss is followed by a compensatory upregulation of mu opioid receptors in the vicinity of LHRH cell bodies. The increment in mu opioid receptors presumably renders the opioid target cells supersensitive to either residual beta-endorphin or other endogenous mu ligands, such as met-enkephalin, thus resulting in chronic opioid suppression of the pattern of LHRH release, and subsequently that of LH. Indeed, prevention of the neuroendocrine effects of estradiol by antioxidant treatment also prevents the cascade of neuroendocrine aberrations resulting in anovulatory acyclicity. The loss of beta-endorphin neurons along with the paradoxical opioid supersensitivity which ensues, provides a unifying framework in which to interpret the diverse features that characterize the reproductively senescent female.

Aging↗

PubMedDate not supplied

Effect of tumor necrosis factor alpha and beta on human oligodendrocytes and neurons in culture.

Cytokines produced by infiltrating hematogenous cells or by glial cells activated during the course of central nervous system disease or trauma are implicated as mediators of tissue injury. In this study, we have assessed the extent and mechanism of injury of human-derived CNS oligodendrocytes and neurons in vitro mediated by the cytokines tumor necrosis factor alpha and beta and compared these with the tumor necrosis factor independent effects mediated by activated CD4+ T-cells. We found that activated CD4+ T-cells, but not tumor necrosis factor alpha or beta, could induce significant release of lactate dehydrogenase, a measure of cell membrane lysis, from oligodendrocytes within 24 hr. Neither induced DNA fragmentation as measured using a fluorescence nick-end labelling technique. After a more prolonged time period (96 hr), tumor necrosis factor alpha did induce nuclear fragmentation changes in a significant proportion of oligodendrocytes without increased lactate dehydrogenase release. The extent of DNA fragmentation was comparable to that induced by serum deprivation. Tumor necrosis factor beta effects were even more pronounced. In contrast to oligodendrocytes, the extent of DNA fragmentation, assessed by propidium iodide staining, induced in neurons by tumor necrosis factor alpha was less than that induced by serum deprivation. In-situ hybridization studies of human adult glial cells in culture indicated that astrocytes, as well as microglia, can express tumor necrosis factor alpha mRNA.

Adult↗

PubMedDate not supplied

Light microscopic radioautographic localization of somatostatin binding sites in the brainstem of the rat.

The distribution of somatostatin binding sites was investigated by light microscopic radioautography in the brainstem of the rat following in vitro labeling with 125I-Tyr0-DTrp8-somatostatin14. Moderate to high labeling densities were detected within the superior colliculus, the locus coeruleus and subcoeruleus, the parabrachial complex, the nucleus of the solitary tract and the dorsal motor nucleus of the vagus. Most of the white matter was devoid of specific somatostatin binding except for fibers of the glossopharyngeal nerve and the spinal trigeminal tract. In most of the labeled areas, 125I-somatostatin binding was evenly distributed between neuropil and perikarya. In a few instances, however, the binding clearly predominated over nerve cell bodies: namely in the dorsal motor nucleus of the vagus and in the pontine and medullary tegmentum. In the latter two regions, the labeled neurons were identified in adjacent sections by tyrosine hydroxylase immunohistochemistry as belonging to the A5 and A1 catecholamine cell groups, respectively. These findings, together with the confirmed association of somatostatin binding sites with noradrenergic neurons in the locus coeruleus, suggest that interactions with catecholaminergic systems may represent a major mode of action for somatostatin in the brainstem.

Animals↗

PubMedDate not supplied

Electron microscopic autoradiographic localization of opioid receptors in rat neostriatum.

The regional distribution of opioid receptors in the mammalian central nervous system has been extensively studied by light microscopic autoradiography. Little is known, however, about the fine structural localization of these receptor sites apart from the fact that they are present in both synaptosomal and smooth microsomal subcellular fractions of whole brain homogenates. Previously, using the Met-enkephalin analogue FK 33-824 (Sandoz), we determined that brain opioid binding sites could be visualized by electron microscopic autoradiography. We now report on the ultrastructural distribution of these sites in the caudate-putamen of the rat. The vast majority of specifically labelled opioid binding sites were found to be associated with axo-dendritic (53%), axoaxonic (18%) or axo-somatic (3%) neuronal membrane interfaces. Only 7%, however, were associated with synaptic junctions. These results suggest that opioids act primarily at non-junctional interfaces on the dendrites, axons and soma of neurones in the neostriatum.

Animals↗

PubMedDate not supplied

Automated EEG monitoring in defining a chronic epilepsy model.

There has been a recent surge of interest in chronic animal models of epilepsy. Proper assessment of these models requires documentation of spontaneous seizures by EEG, observation, or both in each individual animal to confirm the presumed epileptic condition. We used the same automatic seizure detection system as that currently used for patients in our institution and many others. Electrodes were implanted in 43 rats before intraamygdalar administration of kainic acid (KA). Animals were monitored intermittently for 3 months. Nine of the rats were protected by anticonvulsants [pentobarbital (PB) and diazepam (DZP)] at the time of KA injection. Between 1 and 3 months after KA injection, spontaneous seizures were detected in 20 of the 34 unprotected animals (59%). Surprisingly, spontaneous seizures were also detected during the same period in 2 of the 9 protected animals that were intended to serve as nonepileptic controls. Although the absence of confirmed spontaneous seizures in the remaining animals cannot exclude their occurrence, it indicates that, if present, they are at least rare. On the other hand, definitive proof of epilepsy is invaluable in the attempt to interpret pathologic data from experimental brains.

Amygdala↗

PubMedDate not supplied

Alpha7 nicotinic receptor subunits are not necessary for hippocampal-dependent learning or sensorimotor gating: a behavioral characterization of Acra7-deficient mice.

The alpha7 nicotinic acetylcholine receptor (nAChR) subunit is abundantly expressed in the hippocampus and contributes to hippocampal cholinergic synaptic transmission suggesting that it may contribute to learning and memory. There is also evidence for an association between levels of alpha7 nAChR and in sensorimotor gating impairments. To examine the role of alpha7 nAChRs in learning and memory and sensorimotor gating, Acra7 homozygous mutant mice and their wild-type littermates were tested in a Pavlovian conditioned fear test, for spatial learning in the Morris water task, and in the prepulse inhibition paradigm. Exploratory activity, motor coordination, and startle habituation were also evaluated. Acra7 mutant mice displayed the same levels of contextual and auditory-cue condition fear as wild-type mice. Similarly, there were no differences in spatial learning performance between mutant and wild-type mice. Finally, Acra7 mutant and wild-type mice displayed similar levels of prepulse inhibition. Other behavioral responses in Acra7 mutant mice were also normal, except for an anxiety-related behavior in the open-field test. The results of this study show that the absence of alpha7 nAChRs has little impact on normal, base-line behavioral responses. Future studies will examine the contribution of alpha7 nAChR to the enhancement of learning and sensorimotor gating following nicotine treatments.

Animals↗