Journal of Neuroscience, Vol 8, 3343-3353, Copyright © 1988 by Society for Neuroscience
Muscarine and t-LHRH suppress M-current by activating an IAP- insensitive G-protein
P Pfaffinger
Department of Physiology and Biophysics, University of Washington School of Medicine, Seattle 98195.
The control of M-current by muscarinic ACh receptors and luteinizing
hormone releasing hormone (LHRH) receptors was studied in dialyzed frog
sympathetic ganglion neurons. M-current was recorded in dialyzed cells
without run-down or changes in its biophysical properties and could be
reversibly suppressed by muscarine and teleost LHRH (t-LHRH). However,
dialysis with internal solutions lacking ATP or substituting with APP(NH)P
caused the loss of M-current, suggesting that dephosphorylation suppresses
the activity of M-channels. M-current over- recovers after agonist addition
and removal to a size 30% larger than control, as if latent channels are
activated during the recovery. Dialysis of cells with the G-protein
activators GTP gamma S, fluoride, and aluminum fluoride causes loss of
M-current. G-protein activation by receptors was confirmed by dialysis with
low concentrations of GTP gamma S in competition with GTP. This prevents
the rapid loss of M- current, but addition of muscarine or t-LHRH caused
irreversible loss of M-current, suggesting that both transmitter receptors
do suppress M- current by activating a G-protein. Suppression of M-current
was not affected by treatment with 0.1 microgram/ml pertussis toxin (IAP)
for 24-48 hr. In addition, based on the lack of IAP-specific labeling of
frog sympathetic neuron membrane proteins, no IAP-sensitive G-proteins are
present in these cells. These results indicate that an IAP- insensitive
G-protein couples muscarinic and LHRH receptors to the suppression of
M-current.