Vs. Lemos et K. Takeda, NEUROPEPTIDE Y-2-TYPE RECEPTOR-MEDIATED ACTIVATION OF LARGE-CONDUCTANCE CA2-SENSITIVE K+ CHANNELS IN A HUMAN NEUROBLASTOMA CELL-LINE(), Pflugers Archiv, 430(4), 1995, pp. 534-540
We have proposed recently that a pertussistoxin-insensitive Ca2+ influ
x stimulated by Y-2-type receptor activation in CHP-234 human neurobla
stoma cells underlies increases in intracellular free Ca2+ concentrati
on ([Ca2+](i)) induced by neuropeptide Y (NPY), which were strictly de
pendent on extracellular Ca2+ and independent of internal Ca2+ stores.
We describe here the actions of NPY in these same cells, using the ac
tivity of Ca2+-activated K+ channels as an indicator of [Ca2+](i). The
elementary slope conductance of these channels was 110 +/- 3 pS (with
an asymmetrical K+ gradient), their activity was greatly increased by
application of ionomycin, and they were reversibly blocked by 1 mM te
traethylammonium (TEA) and 100 nM charybdotoxin. Application of 100 nM
NPY, in the presence but not in the absence of extracellular Ca2+ inc
reased the channel open probability. ATP applied in the absence of ext
ernal Ca2+ caused rises both in chan nel open probability and [Ca2+](i
). Inositol trisphosphate production was stimulated by ATP but not by
NPY. In outside-out patches, NPY increased channel open probability, i
ndicating that NPY-associated Ca2+ influx does not require all the int
racellular machinery present in intact cells. Channel activation by NP
Y was unaffected by the replacement of guanosine 5'-triphosphate (GTP)
by (guanosine 5'-O-(2-thiodiphosphate) (GDP[beta S]), a non-hydrolysa
ble GDP analogue, in the pipette internal solution, consistent with th
e lack of involvement of G-proteins in the coupling of Y-2-type recept
ors to Ca2+ influx in CHP-234 cells.