Different time courses of GTP[gamma-S]-induced exocytosis and current oscillations in isolated mouse pancreatic acinar cells.

Abstract

Exocytosis in isolated mouse pancreatic acinar cells was investigated using the dual-frequency method for measuring membrane capacitance and ionic conductances. Under control conditions, single exo- and endocytotic events could be resolved. The total cell capacitance slightly decreased to 98.7 +/- 0.9% of the initial cell capacitance within 10 min after establishing the whole-cell configuration. When guanosine 5'-O-(3-thiophosphate) (GTP[gamma-S] was added to the patch pipette, stepwise elevations in membrane capacitance occurred and the cell capacitance increased to 106.7 +/- 1.6% within 10 min. Exocytosis was also stimulated by GTP[gamma-S] when a Ca2+-free pipette solution supplemented with 1 to 10 mM ethylenebis(oxonitrilo) tetraacetate (EGTA) was used. Measurement of the DC current component in parallel with AC current analysis was used to isolate components of the Ca2+-dependent Cl- and monovalent cation conductances from the whole-cell conductance. These experiments demonstrate that in GTP[gamma-S]-stimulated pancreatic acinar cells: (1) activation of Cl- currents precedes that of cation currents, and (2) fusion of the zymogen granule membrane with the plasma membrane does not lead to incorporation of active Cl- or nonselective cation channels (>/= 10 pS).