Biomedical subjects

Colin J Barnstable

Publications and source records attributed to Colin J Barnstable.

22 records · Page 2Linked to original sources

PubMed2002

cGMP-induced presynaptic depression and postsynaptic facilitation at glutamatergic synapses in visual cortex.

The mechanisms by which the intracellular messenger cGMP can modulate synaptic efficacy remain poorly understood. Here we report that cGMP, acting through cGMP-dependent protein kinase (PKG), has multiple rapid and reversible effects on synaptic transmission in slices and cultures of rodent visual cortex. Extracellular application of the membrane permeable cGMP analog 8-bromoguanosine-3',5'-cyclic monophosphate (8-Br-cGMP) and the PKG specific activator beta-phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclic monophosphorothioate sp-isomer (Sp-8-Br-PET-cGMPS) reduced stimulus-evoked EPSPs in slices. In cortical cultures, both analogs reduced the frequency of spontaneous EPSCs, but not their amplitude. In both slices and cultures, intracellular perfusion of the postsynaptic neurons with a pseudosubstrate inhibitory peptide specific for PKG had no effect on the reduction in EPSPs and EPSCs, indicating that the inhibition occurred at presynaptic sites. Whole-cell calcium currents in cultured cortical neurons were also reduced by both analogs, which may account for the effect on synaptic release. To determine whether cGMP was also acting at postsynaptic sites, we applied exogenous kainate/AMPA and NMDA to the recorded cells directly. cGMP and its analogs showed little effect on the postsynaptic kainate/AMPA responses but produced a dramatic enhancement of NMDA responses. cGMP-induced NMDA potentiation was prevented by the specific PKG inhibitory peptide infused into the postsynaptic cell. In summary, cGMP, acting through PKG, had depressive presynaptic and facilitatory postsynaptic actions at excitatory synapses in the visual cortex. We suggest that these opposing actions may be useful for altering the balance of synaptic inputs to cortical neurons in ways that enhance signals important for synaptic facilitation and neuronal plasticity.

Animals↗

PubMed2002

Tissue culture studies of retinal development.

Because of a limited number of cell types, a series of well-described cell-type-specific markers and a stereotyped sequence of cell development, the retina has been a valuable model of CNS development. Dissociated and explant cultures have been used to help define some of the requirements for differentiation of each major cell class. In addition to mixed-cell cultures it is now possible to use cell purification or selective growth methods to give cultures of single cell types. Alteration of gene expression by viral infection has proved to be a valuable method to help elucidate developmental pathways.

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PubMed2002

Protective effect of arachidonic acid on glutamate neurotoxicity in rat retinal ganglion cells.

PURPOSE: Low concentrations of excitotoxic agents such as glutamate decrease survival of retinal ganglion cells (RGCs) and may be an important cause of RGC death in a variety of retinal diseases. Arachidonic acid (AA), an intercellular messenger in the central nervous system, has been reported to have multiple effects on glutamate receptors, including an inhibitory effect on non-N-methyl-D-aspartate (NMDA) receptors. The purpose of this study was to test the hypothesis that AA could protect RGCs from glutamate neurotoxicity. METHODS: RGCs were purified from the rat retina on postnatal days 7 and 8 by a modified two-step panning method. Survival of RGCs after exposure to glutamate, with or without AA treatment, was measured after 3 days in culture. To visualize calcium signals, RGCs were loaded with a calcium indicator dye, fluo-3 acetoxymethyl ester, and the fluorescence was measured by laser scanning confocal microscopy. Electrophysiological effects of AA on non-NMDA ionotropic receptors were examined by using whole-cell patch clamp configurations. RESULTS: Incubation of RGCs with 25 microM glutamate caused 60% loss of RGCs. This glutamate neurotoxicity was significantly ameliorated by low concentrations of AA. Concentrations of AA above 10 microM were toxic to RGCs. Calcium imaging showed that glutamate-, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid- (AMPA) and kainate-induced intracellular calcium accumulation in these cells was reduced by AA. Electrophysiological recordings revealed that currents mediated by non-NMDA ionotropic receptors were inhibited by AA in a dose-dependent manner. CONCLUSIONS: Low concentrations of AA can reduce glutamate neurotoxicity to RGCs by the inhibition of non-NMDA ionotropic receptors. These results suggest that endogenous or exogenous AA may be used to protect RGCs from glutamate neurotoxicity and that AA may be one potential treatment for RGC loss in a variety of eye diseases, including glaucoma.

Aniline Compounds↗

PubMedDate not supplied

Molecular aspects of vertebrate retinal development.

The formation of retina from neural plate has been mapped extensively by anatomical and molecular methods. The major cascades of transcription factor expression have been identified, and deficits resulting from transcription factor knockouts are well characterized. There is extensive cross-regulation, both positive and negative, at the transcriptional level between transcription factors and this is vital in the formation of neural compartments. Many transcription factors are important at both early stages of optic cup formation and later stages of terminal differentiation of retinal cell types. The transcription factor cascades can be regulated by extrinsic factors, and some of the intracellular signaling pathways whereby this is achieved have been identified. Defining the quantitative interactions between regulatory molecules will be the next step in understanding this excellent model of vertebrate central nervous system (CNS) development.

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