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Measurement of Ca 2+ transients using simultaneous dual-emission microspectrofluorimetry and electrophysiology in individual pituitary cells
 

Measurement of Ca 2+ transients using simultaneous dual-emission microspectrofluorimetry and electrophysiology in individual pituitary cells

Nathalie GUERINEAU, PATRICE MOLLARD, Nathalie Guerineau, Jacques Audin BERNARD Dufy
Biochemical and biophysical research communications, Vol.164(3), pp.1045-1052
15/11/1989
: 2590186
Cytosolic free calcium concentration, [Ca 2+]i, was monitored in individual pituitary GH3B6 cells, loaded with the fluorescent Ca 2+ indicator indo 1 either by internal perfusion through a patch clamp pipette or by exposure to indo 1 acetoxymethyl ester, with the use of a dual-emission apparatus for microspectrofluorimetry. This system was sensitive enough to allow on-line monitoring of [Ca 2+]i (from the ratio of fluorescent intensities) which could be combined simultaneously with whole-cell patch clamp recordings. The following situations were examined: (a) [Ca 2+]i oscillations due to action potential firing, and (b) rapid transient elevations of [Ca 2+]i triggered by voltage-dependent Ca 2+ current. The results indicate that monitoring of [Ca 2+]i at the single cell level with indo 1 provides a powerful means to study the [Ca 2+]i regulation in pituitary cells, which should be applicable to many other cell types.
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