The development and pharmacological characterization of calcium channel currents in cultured embryonic rat septal...

We characterized the development and pharmacology of Ca(2+) channel currents in NGF-treated embryonic day 21 cultured rat septal cells. Using standard whole-cell voltage clamp techniques, cells were held at -80 mV and depolarized to construct current-voltage relations in conditions that eliminated Na(+) or K(+) currents. Barium (10 mM) was used as the charge carrier. Maximum current was produced when cells were depolarized to 0 or +10 mV. Recordings from 77 cells revealed that Ca(2+) channel current density increases over time in culture from nearly 0 pA/pF on day 2 in vitro (0.65+/-0.65 pA/pF) to (6.95+/-1.59 pA/pF) on days 6-8. This was followed by a period where currents became nearly 3 times more dense (21.05+/-7.16 pA/pF) at days 9-17. There was little or no evidence for low voltage activated currents. Bath application of 50-100 microM CdCl(2) abolished approximately 95% of the current. Application of 10 microM nimodipine produced a 50.5+/-3.22% reduction in current, 2 microM omega-CTx-GVIA produced a 32.4+/-7.3% reduction, and application of 4 microM omega-Aga-IVA produced a 29.5+/-5.73% reduction in current. When all three inhibitors (10 microM nimodipine, 2 microM omega-CTx-GVIA, and 4 microM omega-Aga-IVA) were applied simultaneously, a residual current remained that was 18.0+/-4.9% of the total current and was completely abolished by application of CdCl(2). This is the first report to characterize Ca(2+) channel currents in cultured embryonic septal cells. These data indicate that there is a steady increase in Ca(2+) channel expression over time in vitro, and show that like other cultured neuronal cells, septal cells express multiple Ca(2+) channel types including L, N, P/Q and R-type channels.