Brain research. Molecular brain research
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Brain Res. Mol. Brain Res. · May 1995
Reduced electrical excitability of PC12 cells deficient in GAP-43: comparison with GAP-43-positive cells.
The electrical excitability of 3 lines of rat pheochromocytoma (PC12) cells were determined under current-clamp recording conditions. In the presence of nerve growth factor (NGF), PC12(A) 'control' cells expressed high levels of GAP-43 protein, PC12(B) cells were highly deficient in GAP-43, and PC12(AB) cells, created by transfection of PC12(B) cells with a rat GAP-43 gene construct, expressed high levels of GAP-43. ⋯ These spikes were resistant to TTX, were greatly enhanced in TEA and TTX, and were substantially reduced by L-type Ca(2+)-channel antagonists. GAP-43 expression may regulate PC12 cell excitability following NGF treatment, as reflected in a lower proportion of cells capable of discharging with Ca(2+)-spikes in a GAP-43-deficient cell line.
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Brain Res. Mol. Brain Res. · Apr 1995
Alterations in tyrosine hydroxylase expression following partial lesions of the nigrostriatal bundle.
Effects of destruction of central dopaminergic neurons on tyrosine hydroxylase gene expression were investigated. Two weeks after the unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle, a 67% to 99% loss of striatal dopamine (DA) content was observed ipsilateral to the injection site. Measures of tyrosine hydroxylase (TH) protein levels revealed losses in striatal content proportional to DA content. ⋯ The decreases in cross-sectional area and TH mRNA content resulted in a small decrease in TH mRNA density of 6%. The determination of TH transcription rate by an intron-directed in situ hybridization assay found no significant change in TH transcriptional activity as a function of lesion. We conclude that the short-term effect of partial 6-OHDA-induced lesions of the nigrostriatal dopaminergic pathway is the selective loss or shrinkage of large DA neurons of the SNc, and that the associated down-regulation of TH mRNA expression in surviving neurons is due to a post-transcriptional mechanism related either to concomitant cellular hyperactivity or is secondary to the morphological alterations.
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Brain Res. Mol. Brain Res. · Oct 1994
Expression of the growth-associated protein B-50/GAP43 via a defective herpes-simplex virus vector results in profound morphological changes in non-neuronal cells.
This study describes the creation and application of a defective herpes simplex viral (HSV) vector for B-50/GAP-43, a neural growth-associated phosphoprotein. We demonstrate abundant expression of B-50/GAP-43 in cultured non-neuronal cells (African green monkey kidney cells [vero cells] and Rabbit skin cells) via this HSV vector. When B-50/GAP-43 was expressed in non-neuronal cells major morphological changes occurred that included extensive membrane ruffling, the formation of filopodia and long thin extensions reminiscent of neurites. ⋯ Some immunoreactivity was associated with vesicular structures that appear to be in-transit in the processes. These observations suggest that B-50/GAP-43 acts at the plasmamembrane to induce a neuron-like morphology in non-neuronal cells persisting for several days in culture. In the future the defective viral vector will enable gene transfer to express B-50/GAP-43 in neurons in vivo in order to study its involvement in regenerative sprouting and neuroplasticity.
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Brain Res. Mol. Brain Res. · May 1994
Role of a 35 kDa fos-related antigen (FRA) in the long-term induction of striatal dynorphin expression in the 6-hydroxydopamine lesioned rat.
D1 dopamine (DA) receptor agonists induce the expression of the opioid peptide dynorphin (DYN) in the striatum, an effect accentuated several fold by removing the dopaminergic innervation to the striatum (e.g., by lesioning the DA cell bodies in the substantia nigra [SN]). D1 receptor-mediated effects are thought to involve cAMP and/or phosphoinositides as second messengers. However, it is unclear what third messengers are involved in the regulation of DYN expression. ⋯ Consistent with the notion that Fos and FRA proteins alter transcriptional activity by binding to AP-1 (or AP-1-like) DNA sequences in the promoter regions of target genes, we found that repeated APO treatment caused large increases in AP-1 binding activity in striata ipsilateral to 6-OHDA lesions. These data indicate that repeated activation of D1 receptors increases both the expression of a 35 kDa FRA and AP-1 binding, events which may mediate the large increases in DYN expression in the DA denervated striatum. While co-administration of the NMDA receptor antagonist, MK-801, inhibited APO-induced increases in DYN and Fos/FRA expression in the intact striatum, its only effect in the DA-denervated striatum was a partial (35%) inhibition of the APO-induced increase in DYN-ir concentrations.(ABSTRACT TRUNCATED AT 400 WORDS)
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Brain Res. Mol. Brain Res. · Apr 1993
Comparative StudyRegion-specific expression of subunits of ionotropic glutamate receptors (AMPA-type, KA-type and NMDA receptors) in the rat spinal cord with special reference to nociception.
The present study attempted to explore the gene expression of the subunits (GluR1-4) of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type receptor, subunit (GluR5) of kainic acid (KA)-type receptor, NR1 [a subunit of N-methyl-D-aspartate (NMDA) receptors] and the possible glutamate-binding subunit of an NMDA receptor complex in the dorsal horn of the rat spinal cords using in situ hybridization histochemistry. These results were compared with those of the spinal motor neurons. Expression of the subunits of the AMPA-type receptor was also examined at the protein level using immunocytochemistry, with reference to the motor neurons. ⋯ Expression of GluR5 is also low in the motor neurons. However, expression of NR1 and the glutamate-binding subunit of an NMDA receptor complex is very strong. These findings indicate that the subunit composition of the AMPA-type receptors regulating motor neurons is different from that of the AMPA-type receptors in the spinal sensory neurons, and that there are at least two kinds of glutamergic systems which regulate motor neurons: via AMPA-type receptors and via NMDA receptors.