Multiple components of both transient and sustained barium currents in a rat dorsal root ganglion cell line.
1. Currents through voltage-activated Ca2+ channels in rat dorsal root ganglion (DRG) x mouse neuroblastoma hybrid (F-11) cells were studied using the whole-cell patch clamp technique with 30 mM-Ba2+ as charge carrier. Two components of the inward Ba2+ current were distinguished on the basis of voltage dependence and time course. Each component could be further subdivided based on pharmacology. 2. A transient inward current activated at test potentials positive to -40 mV, peaked within 20 ms and then decayed during a 200 ms depolarization. The peak amplitude of the transient current occurred between -10 and +10 mV. With a 300 ms conditioning pulse, half-inactivation of the transient current occurred at -30 mV. A sustained inward current activated at test potentials positive to -30 mV and reached a maximum at +20 to +30 mV. The sustained current showed little voltage-dependent inactivation over 200 ms. The amplitudes of both the transient and sustained currents were increased by perfusing with Ba2+ instead of Ca2+. 3. Most F-11 cells had both the transient and sustained Ba2+ currents although the relative amount of the two currents varied with culture conditions. The transient current was more prominent in cells fed with a 'growth' medium (15-20% serum) whereas the sustained current was increased in cells fed with a 'differentiation' medium (1% serum plus growth factors). F-11 cells can be used to study transient current in relative isolation from sustained Ca2+ current under certain culture conditions. The neuroblastoma parent of the F-11 cell line, N18TG-2 cells, exhibited little or no voltage-dependent Ba2+ current. 4. Brief application of omega-conotoxin fraction GVIA (10 microM) produced a long-lasting block of 81% of the sustained current and 27% of the transient current. 5. The transient and sustained Ba2+ currents in F-11 cells were reversibly blocked by brief exposure to Cd2+ or Ni2+. Block of the sustained current was evident with 100 nM-Cd2+ whereas the threshold concentration for Ni2+ block was 1 microM. Cd2+ and Ni2+ were equipotent blockers of the transient current. Dose-response curves for Cd2+ and Ni2+ block of both sustained and transient currents had shallow slopes suggesting that the block was more complex than a simple bimolecular interaction between blocker and one blocking site. Dose-response curves were fitted by a model that included two binding sites for each divalent blocker.(ABSTRACT TRUNCATED AT 400 WORDS)
Boland
L M
LM
Department of Pharmacology, University of North Carolina, School of Medicine, Chapel Hill 27599-7365.
Dingledine
R
R
D000818:Animals; D001464:Barium / Q000378:metabolism*; D002104:Cadmium / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism*; D002460:Cell Line; D004095:Dihydropyridines / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000378:metabolism*; D008564:Membrane Potentials; D008978:Mollusk Venoms / Q000494:pharmacology; D009532:Nickel / Q000494:pharmacology; D009553:Nimodipine / Q000494:pharmacology; D051381:Rats; D013997:Time Factors; D020866:omega-Conotoxin GVIA
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D004095:Dihydropyridines; D008978:Mollusk Venoms; D002104:Cadmium; D001464:Barium; D009553:Nimodipine; D009532:Nickel; D020866:omega-Conotoxin GVIA