title: Cav1.4 encodes a calcium channel with low open probability and unitary conductance.
abstract: When transiently expressed in tsA-201 cells, Ca(v)1.4 calcium channels support only modest whole-cell currents with unusually slow voltage-dependent inactivation kinetics. To examine the basis for this unique behavior we used cell-attached patch single-channel recordings using 100 mM external barium as the charge carrier to determine the single-channel properties of Ca(v)1.4 and to compare them to those of the Ca(v)1.2. Ca(v)1.4 channel openings occurred infrequently and were of brief duration. Moreover, openings occurred throughout the duration of the test depolarization, indicating that the slow inactivation kinetics observed at the whole-cell level are caused by sustained channel activity. Ca(v)1.4 and Ca(v)1.2 channels displayed similar latencies to first opening. Because of the rare occurrence of events, the probability of opening could not be precisely determined but was estimated to be <0.015 over a voltage range of -20 to +20 mV. The single-channel conductance of Ca(v)1.4 channels was approximately 4 pS compared with approximately 20 pS for Ca(v)1.2 under the same experimental conditions. Additionally, in the absence of divalent cations, Ca(v)1.4 channels pass cesium ions with a single-channel conductance of approximately 21 pS. Although Ca(v)1.2 opening events were best described kinetically with two open time constants, Ca(v)1.4 open times were best described by a single time constant. BayK8644 slightly enhanced the single-channel conductance in addition to increasing the open time constant for Ca(v)1.4 channels by approximately 45% without, however, causing the appearance of an additional slower gating mode. Overall, our data indicate that single Ca(v)1.4 channels support only minute amounts of calcium entry, suggesting that large numbers of these channels are needed to allow for significant whole-cell current activity, and providing a mechanism to reduce noise in the visual system.
authors.affiliation: Hotchkiss Brain Institute, Department of Physiology and Biophysics, Faculty of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, Alberta T2N 4N1, Canada.
authors.forename: Clinton J
authors.initials: CJ
authors.lastname: Doering
authors.forename: Jawed
authors.initials: J
authors.lastname: Hamid
authors.forename: Brett
authors.initials: B
authors.lastname: Simms
authors.forename: John E
authors.initials: JE
authors.lastname: McRory
authors.forename: Gerald W
authors.initials: GW
authors.lastname: Zamponi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001703:Biophysics / Q000379:methods*; D015220:Calcium Channels / Q000737:chemistry; D020746:Calcium Channels, L-Type / Q000737:chemistry; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004594:Electrophysiology; D018698:Glutamic Acid / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D011336:Probability; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014162:Transfection
chemical_list: C113330:CACNA1F protein, human; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D018076:DNA, Complementary; C419385:L-type calcium channel alpha(1C); D018698:Glutamic Acid; D012313:RNA; D004247:DNA