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[γ-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[γ-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[γ-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).
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Schmid, A., Schulz, I. Different time courses of GTP[γ-S]-induced exocytosis and current oscillations in isolated mouse pancreatic acinar cells. Pflügers Arch. 432, 876–884 (1996). https://doi.org/10.1007/s004240050211
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DOI: https://doi.org/10.1007/s004240050211