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E A Neale

Publications and source records attributed to E A Neale.

51 records · Page 3Linked to original sources
PubMed1981

Effects of phenytoin on [3H]diazepam binding in dissociated primary cortical cell culture.

The effects of chronic exposure of primary dissociated cerebral cortical cells in culture to the anticonvulsant drug phenytoin have been investigated using benzodiazepine binding techniques. By separating benzodiazepine binding into pharmacologically distinct subtypes, the data indicate that clonazepam-displaceable benzodiazepine binding (associated primarily with neuronal membranes) is significantly decreased by exposure to therapeutic and toxic doses of phenytoin while R05-4864-displaceable benzodiazepine binding (associated principally with non-neuronal elements) is enhanced. The ratio of clonazepam-displaceable to R05-4864-displaceable benzodiazepine binding appears to be the most sensitive indicator for these changes.

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PubMed1978

Intracellular horseradish peroxidase injection for correlation of light and electron microscopic anatomy with synaptic physiology of cultured mouse spinal cord neurons.

Synaptic interactions between spinal cord neurons grown in dissociated cell culture were studied electrophysiologically, and presynaptic cells were subsequently injected by intracellular iontophoresis with horseradish peroxidase (HRP). Following histochemical processing, injected cells were filled with dense reaction product which facilitated the light and electron microscopic identification of the individual physiologically typed neurons. This technique applied to neurons in monolayer culture allowed the visualization of complex intercellular relationships in essentially two dimensions. The number and distribution of morphologically defined synaptic contacts was determined for correlation with individual evoked postsynaptic potentials. HRP-filling of inhibitory and excitatory neurons revealed differences with respect to cellular geometry, axonal projection, and the number, location and ultrastructure of synaptic contacts.

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PubMed1976

Protein metabolism in transected peripheral nerves of the crayfish.

The significance of the protein metabolism in crayfish peripheral nerve was studied in relation the ability of crayfish motor axons to survive for over 200 days following axotomy. In contrast to frog peripheral nerves, the crayfish nerves appear to more closely resemble ganglia in their profiles of synthesis expressed on sodium dodecyl sulfate (SDS) gels, and have higher incorporation rates of [3H]leucine into protein than ganglia. Since anisomycin inhibits over 95% of protein synthesis in crayfish peripheral nerve, it was concluded that this local protein synthesis was dependent upon a eukaryotic ribosomal mechanism. Radioautography of isolated nerves reveals newly synthesized proteins in glial sheaths, and also within the axoplasm of large motor fibers. Based upon the data available at present, a hypothesis that the glia surrounding the axons are responsible for the local protein synthesis, and that some of these newly synthesized proteins are transported into the axon, is presented. Transection of crayfish peripheral nerves proximal to the neuron cell bodies produced a more than two-fold increase in [3H]leucine incorporation, but no significant changes in labeling profiles of the proteins on SDS gels. The data suggest that while an active local protein synthesis may be necessary for the maintenance of several crayfish motor axons, it is not a sufficient condition.

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PubMed1975

Localized catecholamine storage associated with granules in murine neuroblastoma cells.

Catecholamine storage was examined in cultures of the murine neuroblastoma cell line, N-TD6, using histofluorescence, electron microscopic, isotopic and radioautographic criteria. This line was originally derived from uncloned, C1300 tumor cells by selection in tyrosine deficient medium. N-TD6 cells possess both tyrosine hydroxylase (tyrosine-3-monooxygenase, EC 1.14.16.2) and dopamine beta-hydroxylase (dopamine beta-monooxygenase, EC 1.14.17.1) activities. When examined for paraformaldehyde-induced histofluorescence, a small percentage of cells in the population show intense catecholamine fluorescence, often localized within discrete regions of the cellular processes. Electron microscopic examination of these cells reveals both electron lucent vesicles and more frequent, electron dense granules, 50-70 nm and 100-300 nm in diameter, respectively. The distribution of these granules and vesicles varies, but they appear most numerous near the cell surface, along processes and within process endings. By labeling cells with [3H]dopamine and then allowing the cells to release unbound label in the presence of unlabeled dopamine, the localization of catecholamine stores was visualized by radioautographic techniques. While a variety of intracellular distribution of radioactivity were observed, the most prominent concentrations were found in the processes and their terminals; no labeled material was retained when reserpine was present during uptake. The topographic coincidence of granules, catecholamine fluorescence and [3H]dopamine retention in these neuroblastoma cells suggests that catecholamines are stored within these granules in a manner analogous to that observed in normal adrenergic neurons.

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PubMed1975

Evaluation of [3-H]proline for radioautographic tracing of axonal projections in the teleost visual system.

The efficacy of [3-H]proline radioautography for tracing retinal ganglion cell projections to the optic tectum of the jewel fish, Hemichromis bimaculatus, has been compared with that of degeneration techniques. There was good agreement between the various methods. Retinal projections to the optic tectum of two other teleosts, the oscar, Astronotus ocellatus, and the goldfish, Carrasius auratus, were examined radioautographically. In addition to conventional methods of analysis, radioautograms were scanned in a slit microdensitometer and by an automated isodensity scanning system. Results of studies with the protein synthesis inhibitor, cycloheximide, are compatible with the suggestion that axonally transported proteins labeled with [3-H]proline may release diffusible precursors that are reincorporated into protein in adjacent regions. The possible advantages and limitations of radioautography of [3-H]proline-labeled axonally transported protein in brief or extended studies are discussed in terms of the results obtained in the teleost visual system.

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PubMed1972

Radioautography of the optic tectum of the goldfish after intraocular injection of ( 3 H)proline.

Radioautography of the optic tectum of the goldfish, performed after injection of [(3)H]proline into the contralateral eye, effectively resolves several distinct layers of retinal synapses. Silver grains are found unilaterally over nerve tracts containing efferent fibers from the tectum, a result that suggests intercellular migration of labeled molecules. The low background and high specific grain density obtained with [(3)H]proline radioautography indicate the usefulness of this technique for the elucidation of neuroanatomical connections in the visual system.

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PubMedDate not supplied

Differential toxicity of chronic exposure to phenytoin, phenobarbital, or carbamazepine in cerebral cortical cell cultures.

The effects of phenytoin (30 micrograms/ml), phenobarbital (64 micrograms/ml), and carbamazepine (24 micrograms/ml) were assessed in cerebral cortical cell cultures. After antiepileptic drug exposure for eleven days, cultures were assayed for total protein, number of neurons, tetanus toxin fixation, high-affinity uptake of gamma-aminobutyric acid and beta-alanine, activity of choline acetyltransferase, and benzodiazepine binding. Carbamazepine-exposed cultures demonstrated minimal effects, whereas highly significant deficits related to generalized toxicity were observed in cultures exposed to phenytoin or phenobarbital.

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PubMedDate not supplied

Differential neurochemical effects of chronic exposure of cerebral cortical cell culture to valproic acid, diazepam, or ethosuximide.

We have assessed the relative neurochemical effects of valproic acid, ethosuximide, and diazepam on dissociated cultures of mouse cerebral cortex. Cultures were exposed chronically (11 days) to each antiepileptic drug and assayed for number of neurons, total protein, tetanus toxin fixation, high-affinity uptake of gamma-aminobutyric acid and beta-alanine, choline acetyltransferase activity, and specific and clonazepam-displaceable benzodiazepine binding. Ethosuximide-exposed cultures did not evidence neuronal toxicity; exposure to valproic acid and diazepam resulted in modest neuronal toxicity. However, exposure to each of these drugs resulted in a marked reduction in benzodiazepine binding. This effect may relate to a common mechanism of action of drugs used to treat absence seizures.

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