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  • Title: Activation of glutamate receptors and glutamate uptake in identified macroglial cells in rat cerebellar cultures.
    Author: Wyllie DJ, Mathie A, Symonds CJ, Cull-Candy SG.
    Journal: J Physiol; 1991 Jan; 432():235-58. PubMed ID: 1653320.
    Abstract:
    1. Patch-clamp methods have been used to examine the action of excitatory amino acids on three types of glial cell in cultures of rat cerebellum, namely type-1-like astrocytes, type-2 astrocytes and oligodendrocytes. In addition we have examined glutamate sensitivity of the precursor cell (the O-2A progenitor) that gives rise to type-2 astrocytes and oligodendrocytes. 2. Glutamate (30 microM), quisqualate (3-100 microM), (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA, 10-30 microM) and kainate (10-500 microM) were applied to cerebellar type-2 astrocytes examined under whole-cell voltage clamp. Each of these agonists induced inward currents in cells held at negative membrane potentials. The currents reversed direction near 0 mV holding potential. N-Methyl-D-aspartate (NMDA, 30-100 microM) or aspartate (30 microM) in the presence of glycine (1 microM) did not evoke any whole-cell current changes in type-2 astrocytes. 3. The distribution of glutamate receptors in type-2 astrocytes was mapped with single- or double-barrelled ionophoretic pipettes containing quisqualate or kainate. Application of these agonists (current pulses 100 ms, 50-100 nA) to cells held at -60 mV evoked inward currents of 20-120 pA in the cell soma and 10-80 pA in the processes. Responses could also be obtained at the extremities of processes (approximately 60 microns from the soma). 4. Quisqualate or kainate (at 30 microM) applied to O-2A progenitor cells from rat cerebellum or optic nerve induced whole-cell currents (quisqualate 20-30 pA; kainate 20-50 pA, holding potential, Vh = -60 mV) that reversed near 0 mV. In common with type-2 astrocytes, the progenitor cells did not respond to NMDA (30 microM). 5. Type-1-like astrocytes produced large inward currents to glutamate (30 microM). These currents remained inward-going at holding potentials as positive as +80 mV and were not accompanied by any apparent noise increase. This result can be explained by the presence of an electrogenic glutamate uptake carrier. In cells kept up to 4 days in vitro, quisqualate, kainate and NMDA each failed to produce any whole-cell current changes, indicating the absence of receptors in type-1-like astrocytes at this stage in culture. Furthermore the glutamate uptake currents in type-1-like astrocytes were inhibited when external Na+ was replaced by Li+, although Li+ was found to pass through the glutamate channel in type-2 astrocytes.(ABSTRACT TRUNCATED AT 400 WORDS)
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