| Abstrakt: |
Crude nerve-ending preparations from rat brain were labeled with either [(32)P]phosphate or myo[2-(3)H]inositol in order to observe effects of guanosine 5?-[?-thio]triphosphate (GTP?S) and other nucleotides on phosphoinositides, phosphatidate and inositol phosphates. This system exhibited typical responses to muscarinic agonists, including acetylcholine-a decrease in net labeling of [(32)P]polyphosphoinositides, an increase in labeling of [(32)P]phosphatidate, and a stimulation of [(3)H]inositol phosphate formation. GTP?S and other nucleotides may not readily penetrate intact synaptosomal membranes to cause activation of phospholipase C via an interaction with G proteins, and, as might be expected, there was no indication that G-protein interaction occurred in these preparations. However, other effects were observed. GTP?S decreased net [(32)P] incorporation in phosphatidylinositol (PI) and polyphosphoinositides in a dose-dependent manner. GTP?S also caused an initial marked stimulation of [(32)P] labeling of phosphatidate, suggesting a possible inhibition in the conversion of phosphatidic acid to PI. Other nucleotides [GTP, ATP, Gpp(NH)p, GMP] produced qualitatively similar effects on phosphoinositides. Thus GTP?S and other nucleotides, at physiologically relevant concentrations, may influence phosphoinositide turnover via extracellular or other mechanisms, in addition to the proposed interaction of GTP with G-proteins within membranes. |
