Calcium-induced inactivation of calcium current causes the inter-burst hyperpolarization of Aplysia bursting neurones.
A triphasic series of tail currents which follow depolarizing voltage-clamp pulses in Aplysia neurones L2-L6 was described in the preceding paper (Kramer & Zucker, 1985). In this paper, we examine the nature of the late outward component of the tail current (phase III) which generates the inter-burst hyperpolarization in unclamped cells. The phase III tail current does not reverse between -30 and -90 mV, and is relatively insensitive to the external K+ concentration. In contrast, Ca2+-dependent K+ current (IK(Ca)), elicited by intracellular Ca2+ injection, reverses near -65 mV, and the reversal potential is sensitive to the external K+ concentration. Addition of 50 mM-tetraethylammonium (TEA) to the bathing medium causes a small increase in the phase III tail current. In contrast, IK(Ca) is completely blocked by addition of 50 mM-TEA. The phase III tail current is suppressed by depolarizing pulses which approach ECa, is blocked by Ca2+ current antagonists (Co2+ and Mn2+), and is blocked by intracellular injection of EGTA. The phase III tail current is reduced by less than 10% after complete removal of extracellular Na+. These bursting neurones have a voltage-dependent Ca2+ conductance which exhibits steady-state activation at a membrane potential similar to the average resting potential of the unclamped cell (i.e. -40 mV). The steady-state Ca2+ conductance can be inactivated by Ca2+ injection, or by depolarizing pre-pulses which generate a large influx of Ca2+. The steady-state Ca2+ conductance has a voltage dependence similar to that of the phase III tail current. The Ca2+-dependent inactivation of the steady-state Ca2+ conductance occurs in parallel with the phase III tail current; both have a similar sensitivity to Ca2+ influx, and both processes decay with similar rates after a depolarizing pulse. Hence, we propose that the phase III tail current is due to the Ca2+- dependent inactivation of a steady-state Ca2+ conductance. The decay of IK(Ca) following simulated spikes or bursts of spikes is rapid (less than 1 s) compared to the time course of the phase III tail current and the inter-burst hyperpolarization (tens of seconds). Thus, we conclude that IK(Ca) does not have a major role in terminating bursts or generating the inter-burst hyperpolarization in these cells. We present a qualitative model of the ionic basis of the bursting pace-maker cycle. The central features of the model are the voltage-dependent activation and the Ca2+-dependent inactivation of a Ca2+ current.
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Kramer
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Zucker
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D001048:Aplysia / Q000502:physiology*; D002118:Calcium / Q000494:pharmacology / Q000502:physiology*; D066298:In Vitro Techniques; D007473:Ion Channels / Q000187:drug effects*; D008345:Manganese / Q000494:pharmacology; D008959:Models, Neurological; D009474:Neurons / Q000502:physiology*; D011188:Potassium / Q000494:pharmacology; D019789:Tetraethylammonium; D013757:Tetraethylammonium Compounds / Q000494:pharmacology; D013997:Time Factors
D007473:Ion Channels; D013757:Tetraethylammonium Compounds; D008345:Manganese; D019789:Tetraethylammonium; D011188:Potassium; D002118:Calcium