1. Changes in intracellular Ca2+ concentration ([Ca2+]1) induced by activation of GABAA receptors (synaptic stimulation or application of the GABAA agonist isoguvacine) were studied on pyramidal cells and interneurons from hippocampal slices of rats from two age groups (postnatal days (P) 2-5 and P12-13) using the fluorescent dye fluo-3 and a confocal laser scanning microscope. Cells were loaded with the dye either intracellularly, using patch pipettes containing fluo-3 in the internal solution, or extracellularly, using pressure pulses applied to an extracellular pipette containing the permeant dye fluo-3 AM. 2. Interneurons and pyramidal cells from P2-5 slices loaded with fluo-3 AM responded by an increase in [Ca2+]i to isoguvacine and to glutamate, in contrast to cells from P12-13 slices which responded to glutamate but not to isoguvacine. In the mammalian adult central nervous system, GABA postsynaptically localized GABAB receptors activate (y-aminobutyric acid) is the main inhibitory neuro-potassium conductances or decrease calcium currents transmitter, acting on two main classes of

We report here the technique for detection and measurement of the temperature changes in single cells using a recently devised microthermometer (a glass micropipette filled with the thermosensitive fluorescent dye Europium (III) thenoyltrifluoroacetonate trihydrate). We found that the heat production in a single HeLa cell occurred with some time delay after the ionomycin-induced Ca(2+) influx from the extracellular space. The time delay inversely depended on extracellular [Ca(2+)], and the increase in temperature was suppressed when Ca(2+)-ATPases were blocked by thapsigargin. These observations strongly suggest that the enzymatic activity of Ca(2+)-ATPases in endoplasmic reticulum leads to the heat production. This study has therefore paved the way for studying the thermogenesis at the single-cell level.

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