[26] Using confocal microscopy and the fluorescent indicator, 6-methoxy-N-ethylquinolinium (cas 134907-10-1) iodide, to measure changes in intracellular chloride

Add time:08/04/2019    Source:sciencedirect.com

Publisher SummaryThe chapter presents approaches of studying dynamic changes in intracellular Cl– in living neurons with the laser scanning confocal microscope (LSCM) and a fluorescent Cl– indicator, 6-methoxy-N-ethylquinolinium (cas 134907-10-1) iodide (MEQ). The chapter discusses the brain slice preparation. Brain slices of the hippocampus (containing somatosensory cortex) or cerebellum are prepared from rats between 8 and 21 days old. All laser scanning confocal microscopes have an improved signal-to-noise ratio and the ability to penetrate into tissues, although slit variations. The Cl– channels in the areas studied, include the recently discovered ClC-2 family as well as GABAA (γ-aminobutyric acid) receptors; activation of the latter in adult animals leads to neuronal inhibition via a stimulated Cl– conductance. Applying MEQ and confocal microscopy to the brain slice preparation, an appropriate GABAA receptor pharmacology is demonstrated. In addition, a comparison of simulated ischemia versus activation of excitatory amino acid receptor subtypes identified different mechanisms for Cl– influx and reduced GABAergic function. The presence of cell swelling can be investigated with fluorescence microscopy or microfluorimetry with the volume-sensitive fluorescent dye, calcein-AM.

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