The involvement of inositol 1,4,5-trisphosphate and calcium in the two-component response to acetylcholine in Xenopus oocytes

Abstract

1. The membrane response to acetylcholine (ACh), inositol 1,4,5-trisphosphate (IP3) and intracellular Ca2+ was studied in Xenopus laevis oocytes under voltage-clamp conditions. 2. Shallow, submembranal injections of IP3 in the animal hemisphere of the oocyte evoked a two-component response comprised of a rapid, transient component followed by a slow, sustained component. 3. When the injection pipette was inserted further into the cell (to 300 microns below the cell membrane), the fast component diminished and the slow component remained unchanged or even increased. 4. The rapid component exhibited an apparent higher sensitivity to IP3 compared to the slow component. 5. The two components of the IP3 response were retained in a Ca2+-free environment. 6. Injection of a single large dose (20-50 pmol) of CaCl2 into the oocyte evoked a typical two-component response, whereas repetitive threshold doses (0.1 pmol CaCl2) elicited large current fluctuations which developed into a small depolarization current. 7. The delay in the peak of the slow component of the response to either IP3 or to CaCl2 injections appeared too long to be accounted for by diffusion alone. 8. Depletion of oocyte Ca2+ by the divalent cation ionophore A23187 (greater than 1 microM) inhibited the response to ACh and IP3. Low concentrations of A23187 selectively inhibited the rapid component of the ACh response, though not the rapid component of the IP3 response. 9. Our data suggest that the two-component membrane response to ACh in Xenopus oocytes can be accounted for by ACh-induced elevation of IP3 and subsequent IP3-induced release of intracellular Ca2+.