Two types of Ca2+ currents are found in bovine chromaffin cells: facilitation is due to the recruitment of one type
1. Whole-cell Ca2+ currents in cultured bovine chromaffin cells were studied using patch-clamp electrophysiology. With Ba2+ or Ca2+ as the current carriers, two separate components of whole-cell current could be distinguished by biophysical and pharmacological criteria. These components of Ca2+ curr...
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Veröffentlicht in: | The Journal of physiology 1991-01, Vol.432 (1), p.681-707 |
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Zusammenfassung: | 1. Whole-cell Ca2+ currents in cultured bovine chromaffin cells were studied using patch-clamp electrophysiology. With Ba2+
or Ca2+ as the current carriers, two separate components of whole-cell current could be distinguished by biophysical and pharmacological
criteria. These components of Ca2+ current were different from T- or N-type Ca2+ channels previously described, as they were
not inactivated at a holding potential of -60 mV. 2. Depolarization of the cells past -20 mV in 10 mM-Ba2+ activated a single
component of Ca2+ current, called the 'standard' current. This current showed no detectable voltage-dependent inactivation,
but did show marked current-dependent inactivation as steady-state inactivation (H-infinity) plots obtained in the presence
of Ba2+ were quite different from those obtained from Ca2+. 3. In most chromaffin cells large pre-depolarizations or repetitive
depolarizations in the physiological range activated a second component of Ca2+ current called 'facilitation'. Facilitation
was observed with either Ca2+ or Ba2+ as the charge carrier. Recruiting facilitation increased whole-cell currents by an average
of 60%. 4. Pre-pulses to +120 mV lasting 200 ms completely activated facilitation. Pre-pulses longer than 800 ms started to
inactivate facilitation, while pre-pulses longer than 2500 ms completely inactivated this component of Ca2+ current. Because
only outward currents were recorded at +120 mV, it is likely that facilitation inactivated in a voltage-dependent manner.
5. When the extracellular Ba2+ concentration was increased in the range from 2 to 90 mM activation of both facilitation and
standard Ca2+ currents shifted in the depolarizing direction. In 2 mM-Ba2+ facilitation activated at potentials 10 mV more
negative than the standard component, while in 90 mM-Ba2+, facilitation activated at a potential about 10 mV more depolarized
than the standard component. Thus, the voltage sensor for the facilitation Ca2+ current appeared to sense more surface charge
than did the standard Ca2+ current. 6. Tail currents measured at -20 and -30 mV in the absence of facilitation (without pre-pulses)
showed one time constant for current deactivation. Tail currents measured with both facilitation and standard currents activated
showed a significantly slower deactivation rate than that seen with the standard current alone. 7. The dihydropyridine antagonist
nisoldipine (1 microM) completely suppressed the facilitation Ca2+ current even when cells were he |
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ISSN: | 0022-3751 1469-7793 |
DOI: | 10.1113/jphysiol.1991.sp018406 |