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Simultaneous measurement of Ca2+ and pH by laser cytometry using fluo-3 and SNARF-1

✍ Scribed by Toshiko Atsumi; Kenji Sugita; Minoru Kohno; Tsutomu Takahashi; Takao Ueha

Publisher
John Wiley and Sons
Year
1996
Tongue
English
Weight
921 KB
Volume
24
Category
Article
ISSN
0196-4763

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✦ Synopsis

We present a new conwnient method for simultaneous measurement of intracellular calcium concentration ([Caz+]3 and inlracellular pH (pH3 using laser cytometry with a mixture of fho-3 (for [Ca2']3 and SNARF-1 (for pH3, with iso excitation (488 -)-dual emission (530 nm for fluo-3 and > 630 n m for SNARF-1). F3y using this technique, we measured the changes in [Ca2+], and pH, in A431 human epidermoid catcinolzlll cells and HSG human salivary gland cells stimulated by ATP. We found that alkallzation in A-431 cell occurred with the elevation of [a2+],; in contrast, allcabation in HSG cells did not occur at all, even though the ele-vation of [Ca"], was observed. Using BAPTA (a chelating agent of Ca2') and d o r i d e (an inhibitor of the Na+/H+ exchanger), we found that the elevation of pH, requires the elevation of [Ca2'], but that the elevation of [Ca2'], does not always require a rise in pH,. From our results we conclude that elevation of [ a 2 + ] , takes precedence over the elevation of pH, in ATP-stimulated signal traasduction.

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