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  • Title: Extracellular pH, Ca(2+) influx, and response of vascular smooth muscle cells to 5-hydroxytryptamine.
    Author: Nazarov V, Aquino-DeJesus J, Apkon M.
    Journal: Stroke; 2000 Oct; 31(10):2500-7. PubMed ID: 11022085.
    Abstract:
    BACKGROUND AND PURPOSE: Cerebral vascular smooth muscle cells (VSMCs) contract on extracellular pH (pH(o)) increases and relax on pH(o) decreases. These changes in tone are believed to result from changes in [Ca(2+)](i), although the responsible mechanisms are not fully understood. VSMCs also contract in response to 5-hydroxytryptamine (5-HT), which increases [Ca(2+)](i) via both Ca(2+) release and influx. We hypothesized that examining effects of pH(o) decreases on 5-HT-induced [Ca(2+)](i) changes would allow us to identify mechanisms whereby pH(o) influences tone. Accordingly, we compared [Ca(2+)](i) increases in cerebral VSMCs, evoked by 5-HT, with increases evoked by increased pH(o) and examined 5-HT-dependent [Ca(2+)](i) increases at normal and decreased pH(o). METHODS: We monitored [Ca(2+)](i,), using the Ca(2+)-sensitive dye fura 2, in cultured rat cerebral VSMCs obtained by enzymatic digestion of middle cerebral arteries and their branches (passages 1 to 3) grown on glass coverslips and superfused with physiological saline. RESULTS: Increasing pH(o) from 7.3 to 7.8 increased [Ca(2+)](i), and these increases were prevented in Ca(2+)-free solutions. Decreasing pH(o) from 7.3 to 6.9 did not alter [Ca(2+)](i) unless [Ca(2+)](i) was first raised by treatment with 5-HT (10 micromol/L). 5-HT resulted in biphasic [Ca(2+)](i) increases characterized by transient peaks blocked by the Ca(2+)-ATPase inhibitor thapsigargin (10 nmol/L) and prolonged plateaus blocked by the Ca(2+) channel blocker Ni(2+) (1 mmol/L). Acidification did not alter the transient peaks but significantly reduced 5-HT-induced Ca(2+) influx. CONCLUSIONS: We conclude that increasing pH(o) induces Ca(2+) influx in rat cerebral VSMCs and decreasing pH(o) inhibits 5-HT-stimulated Ca(2+) entry but not intracellular Ca(2+) release.
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