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pp.243-250
To find the mechanism underlying the ADP-induced increase in the outward current in ovulated mouse oocytes, we examined changes in voltage-dependent currents using the whole cell voltage clamp technique and the internal perfusion technique. Eggs were collected from the oviduct of superovulated mice with PMSG and hCG. Membrane potential was held at -60 mV (or -80 mV in the case of recording currents) and step depolarizations or hyperpolarizations were applied for 300 ms. By step depolarizations, outward currents comprising steady-state and time-dependent components were elicited. They were generated in response to the positive potential more than 20 mV with severe outward rectification and were blocked by external TEA, a specific channel blocker, suggesting that they be carried via channels. Internally-perused 5 mM ADP gradually increased outward currents (IK) 1 min after perfusion of ADP and reached slowly to maximum (150~170％) 5 min later over the positive potential range, implying that ADP might not be acted directly to the channels. IK were decreased by 5 mM ATP without affecting the steady-state component of outward current. In contrast to the effect of ADP and ATP on IK, both effect of ATP and ADP on inward currents (ICa) could not be detected due to the continuous decrease in current amplitudes with time-lapse ("run-down" phenomena). To check if there is a G protein-involved regulation in the ionic current of mouse oocytes, 1 mM GTP was applied to the cytoplasmic side, and the outward current and inward currents were recorded. ICa was promptly increased in the presence of GTP whereas IK was not changed. from these results, it is concluded that the ATP-dependent regulation is likely linked in the ADP-induced increase in the outward current, and G protein-involved cellular signalling might affect ion channels carrying and in mouse oocytes.
