The firing patterns of rat melanotrophs recorded using the patch clamp technique.
Cell-attached and whole-cell recordings were made from adult rat melanotrophs maintained in vitro by standard cell culture techniques. In cell-attached recordings the cells showed small biphasic currents which reflected spontaneous cell firing. Single channel currents often had distinct relaxations and depolarizing currents through single channels could trigger the discharge of an action potential in the cell; both observations are consistent with the high input resistance (1-10 G omega) measured in the whole-cell configuration. The discharge of action potentials occurring either spontaneously or by current injection was eliminated by tetrodotoxin or by removing Na from the external medium. A Na-dependent plateau depolarization which activated near the spike threshold was also seen. In cells exposed to tetrodotoxin and K-channel blocking agents it was possible to evoke a long-lasting (up to 20 s) action potential which was enhanced and reduced, respectively, by Ba and Cd and thus appeared to reflect currents through voltage-activated Ca channels. Small amplitude Ca-dependent depolarizations could also be evoked at membrane potentials as low as -40 mV. In cell-attached and whole-cell recordings 10 mM Ba caused the discharge of tetrodotoxin-insensitive action potentials prior to a maintained depolarization of the membrane. The low threshold for Ca-dependent depolarizations suggest that Ca influx might occur in these cells even at the resting potential. Additionally, both a Ca current and the current underlying the Na-dependent plateau depolarization may influence the rate of cell firing and in doing so further increase Ca influx through voltage-activated channels.(ABSTRACT TRUNCATED AT 250 WORDS)
Kehl
S J
SJ
MRC Neuroendocrinology Unit, Newcastle General Hospital, Newcastle Upon Tyne, U.K.
McBurney
R N
RN
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D001464:Barium / Q000494:pharmacology; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D008297:Male; D009074:Melanocyte-Stimulating Hormones / Q000378:metabolism*; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D011919:Rats, Inbred Strains
D015220:Calcium Channels; D001464:Barium; D009074:Melanocyte-Stimulating Hormones