Abstract

In the present investigation, sodium fluoride (NaF) was employed to explore the role of guanine nucleotidebinding proteins (G-proteins), protein kinase-C, or cytosolic calcium ([Ca]i) in the regulation of cytosolic pH ([pH]i) in dispersed bovine parathyroid cells, using the pH-sensitive fluorescent dye BCECF. When cells acidified by nigericin in Nafree medium were resuspended in Na-containing buffer, [pH]i returned to basal levels. This recovery was blocked by continued removal of Na+ or the addition of amiloride. NaF (10 mM) increased [32P]phosphate incorporation into phosphatidylinositol bisphosphate, suggesting an increase in phosphatidylinositol bisphosphate turnover. NaF caused an initial acidification, followed by an alkaline recovery in a dose-dependent manner (1†10 mM). Amiloride blocked the NaF-induced alkaline recovery. The protein kinase-C activator phorbol 12-myristate 13-acetate (10-7 M) caused cytosolic alkalinization, while the protein kinase- C inhibitor H7 (6 Ã-10-5 M) significantly inhibited the NaF-induced alkaline recovery. Pertussis toxin (1 μg/nA) did not affect the NaF-induced changes in [pH]i. Removal of extracellular Ca2+ with EGTA blocked the NaF-induced increase in [Ca]i and alkaline recovery. Ionomycin (5 Ã-10-7 M) caused cytosolic alkalinization, but pretreatment with EGTA inhibited the ionomycin-induced cytosolic alkalinization. The present studies clearly demonstrated the presence of an amiloride-sensitive Na+/H+ exchanger in parathyroid cells. Our findings suggest that the NaF-induced cytosolic alkaline recovery was via two complementing pathways: 1) activation of protein kinase-C, followed by stimulation of a Na+/H+ exchanger, and 2) existence of extracellular calcium and/or an increase in [Ca]i.

Original languageEnglish
Pages (from-to)149-156
Number of pages8
JournalEndocrinology
Volume124
Issue number1
DOIs
StatePublished - Jan 1989

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