Rat lactotrophs isolated by fluorescence-activated cell sorting are electrically excitable.
Lactotrophs (prolactin-containing cells) from the anterior pituitary of the adult female rat were labeled by a cell-surface reaction with anti-prolactin antibodies and then isolated by fluorescence-activated cell sorting (FACS). FACS-isolated pituitary cells were maintained in culture 1-9 days, after which their excitable membrane properties were examined using the whole-cell patch recording technique. Comparisons between lactotrophs sorted at different laser-power settings on the FACS showed that although the electrical excitability of the sorted lactotrophs was acutely altered by exposure to high laser power, excitability comparable to unsorted cells was retained by the use of minimal laser power (10 mW). Recordings were made from 67 cells that had been isolated using the low laser power. These cells had resting membrane potentials ranging from -22 mV to -60 mV. More than 80% of the cells responded to depolarizing current injections with regenerative, full-amplitude, overshooting action potentials. Approximately 20% of these cells exhibited spontaneous 20-30 mV fluctuations in the level of resting membrane potential, the majority of which did not overshoot 0 mV. Both the action potentials and the spontaneous fluctuations involved Na+ and Ca2+ ion conductance mechanisms.
D000200:Action Potentials; D000818:Animals; D004594:Electrophysiology; D005260:Female; D005434:Flow Cytometry; D010903:Pituitary Gland, Anterior / Q000166:cytology*; D011388:Prolactin; D051381:Rats; D011919:Rats, Inbred Strains; D013779:Tetrodotoxin / Q000494:pharmacology
D013779:Tetrodotoxin; D011388:Prolactin