Confocal laser scanning microscopy used to monitor intracellular Ca2+ changes in hippocampal CA 1 neurons during energy deprivation

Brain Research
T O Grøndahl, I A Langmoen

Abstract

An increase in intracellular calcium during cerebral ischemia has been proposed as a common final pathway underlying the events leading to neuronal death. Intracellular calcium has been measured with ion selective electrodes during energy deprivation (ED) in hippocampal slices and with fluorescent techniques in neuronal cultures. In the present study, we describe a novel method to visualize and quantify changes in intracellular calcium in brain slices using Confocal Laser Scanning Microscopy (CLSM). CA 1 pyramidal neurons in hippocampal slices were filled by intracellular injection with a 1:2 mixture of the fluorescent dyes Fluo 3 and Fura Red. The neurons were then visualized using CLSM, and the ratio of the fluorescence from each probe used to quantify intracellular calcium concentrations before and during ED. The free intracellular calcium concentration was 60 nM prior to ED and increased to 24 microM during ED. These results demonstrates that CLSM and fluorescent probes can be used in functional neuronal networks in addition to cell cultures as previously described.

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Citations

Apr 18, 2007·Journal of Neurochemistry·Maria Stella LombardiAnnemieke Kavelaars
Jun 17, 2005·Brain : a Journal of Neurology·Morten C MoeIver A Langmoen
Sep 30, 1999·Microscopy Research and Technique·S A Stricker, M Whitaker
Oct 3, 1999·Physiological Reviews·P Lipton
Apr 26, 2003·Journal of Theoretical Biology·L H Pojoga, R H Hilderman

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