Sodium and calcium currents in acutely dissociated neurons from rat suprachiasmatic nucleus.
1. Neurons were acutely dissociated from the suprachiasmatic nucleus (SCN) of adult rats and studied with whole-cell and perforated-patch recordings at room temperature. 2. Acutely dissociated SCN neurons had spherical cell bodies of 12 microns in average diameter. The recorded cells were randomly selected and had either no process (38%), one (41%), two (19%), or three processes (2%). They had a resting potential of about -60 mV, an input resistance of approximately 5 G omega, and a cell capacitance of approximately 7 pF. 3. The dissociated neurons had variable spontaneous firing rates, typically (76%) < 1 Hz. 4. Under current clamp, continuous current injection elicited repetitive action potentials. 1 microM tetrodotoxin (TTX) reduced the amplitudes of the action potentials as well as the firing rate, whereas 200 microM Cd2+ stopped repetitive firing altogether. Action potentials were completely eliminated with Cd2+ and TTX present. These results suggest that both Na+ and Ca2+ contribute to the action potential in these cells. 5. With 200 microM Cd2+ present to block calcium currents, a train of brief depolarizing pulses could still elicit repetitive sodium action potentials, but these became attenuated at stimulating frequencies as low as 1 Hz. 6. Under voltage clamp, the sodium current was activated at about -40 mV and peaked at about -10 mV. It inactivated with a time constant of approximately 0.5 ms at 0 mV, and in steady state the current was half-inactivated at about -60 mV. Recovery of the current from inactivation showed two very different phases with time constants of approximately 30 and 600 ms at -60 mV. The slow phase was probably responsible for the very low firing rate of the sodium action responsible for the very low firing rate of the sodium action potential. 7. In the absence of external sodium, depolarization-activated calcium action potentials were preferentially blocked by 20 microM Cd2+, whereas a posthyperpolarizing depolarizing (or anode break) was preferentially reduced by 100 microM Ni2+. These differential effects hinted at the presence of both low-threshold and high-threshold calcium currents in these cells. 8. Voltage-clamp experiments confirmed the presence of a low-threshold, transient calcium current that was activated by depolarizations above -70 mV. It inactivated with a time constant of approximately 25 ms between -50 and -30 mV. Steady-state inactivation was half-complete at about -90 mV and complete at about -70 mV.(ABSTRACT TRUNCATED AT 400 WORDS)
Huang
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RC
Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
D000200:Action Potentials / Q000502:physiology; D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism*; D002940:Circadian Rhythm / Q000502:physiology; D005260:Female; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects; D008564:Membrane Potentials / Q000502:physiology; D009474:Neurons / Q000378:metabolism*; D018377:Neurotransmitter Agents / Q000378:metabolism; D011188:Potassium / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012964:Sodium / Q000502:physiology; D015222:Sodium Channels / Q000187:drug effects / Q000378:metabolism*; D013493:Suprachiasmatic Nucleus / Q000166:cytology / Q000378:metabolism*
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D018377:Neurotransmitter Agents; D015222:Sodium Channels; D012964:Sodium; D011188:Potassium