Metabolism of polyphosphoinositide was studied in bulk isolated brain cells. Cells were isolated by a rapid method using mechanical disruption followed by molecular seiving and centrifugation. Incorporation of [32Pi]orthophosphate into phosphatidyl inositol-4-phosphate and phosphatidylinositol-4,5 bis-phosphate was optimum at 30 and 60 min, respectively, in the isolated cells. Breakdown studies showed maximum loss of 32Pi after 60 min. Addition of ethanol at and above 10 mM concentration in vitro significantly decreased the incorporation of 32Pi into both phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5 bis-phosphate by the isolated cells. However, the spontaneous breakdown of polyphosphoinositide was not altered in the presence of ethanol in vitro.
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