Patch-clamp analysis of voltage-gated currents in intermediate lobe cells from rat pituitary thin slices.
Ionic currents of hypophyseal intermediate lobe cells were studied using a thin-slice preparation of the rat pituitary in conjunction with conventional and perforated whole-cell patch-clamp recording techniques. A majority (89%) of the cells studied generated Na+, Ca2+ and K+ currents upon depolarizing voltage steps and responded to bath application of gamma-aminobutyric acid (GABA; 20-50 microM) with inward currents (in symmetrical chloride, holding potential -80 mV). A small percentage of cells (11%) did not display inward membrane currents upon depolarization and was unresponsive to GABA. In the first type of cells, Ca2+ and K+ currents were further studied in isolation. Ca2+ tail currents showed a biphasic time course upon repolarization, with time constants and amplitudes of 2.07 +/- 0.29 ms, 123 +/- 22 pA (for the slowly deactivating component) and 0.14 +/- 0.06 ms, 437 +/- 33 pA (for the fast-deactivating component; means +/- SD of n = 4 cells). Slowly and fast-deactivating conductances were half-maximally activated at around -10 mV and +10 mV respectively. Depolarizing voltage steps elicited two types of K+ current, which were separated using a prepulse protocol. A fast-activating, transient component showed half-maximal steady-state inactivation between -65 mV and -45 mV depending on the divalent cation composition of the external solution. Its decay was fitted by single-exponential functions with time constants of 36 +/- 11 ms and 3.9 +/- 0.9 ms at -20 mV and +40 mV respectively (mean +/- SD; n = 4 cells). Whereas the peak current amplitudes of the transient K+ current component remained stable, the amplitude of the second, delayed component increased progressively throughout the course of whole-cell experiments. In cells recorded with the perforated whole-cell technique, bath application of dopamine (10 nM-1 microM) induced large hyperpolarizations from a spontaneous membrane potential of -40 mV, but did not consistently affect the amplitude of the voltage-gated K+ conductances. These data are compared to previous studies using other preparations of the intermediate lobe, and differences are discussed, thus helping to extend our knowledge of electrical excitability of hypophyseal cells.
Schneggenburger
R
R
Departamento de Fisiología y Biofísica, Facultad de Medicina, Universidad de Sevilla, Spain.
López-Barneo
J
J
D000818:Animals; D002463:Cell Membrane Permeability; D004298:Dopamine / Q000502:physiology; D066298:In Vitro Techniques; D007700:Kinetics; D008564:Membrane Potentials; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D011188:Potassium; D004298:Dopamine