Cyclic AMP-dependent protein kinase decreases GABAA receptor current in mouse spinal neurons.
GABA, the major inhibitory neurotransmitter in the mammalian brain, binds to GABAA receptors, which form chloride ion channels. The predicted structure of the GABAA receptor places a consensus phosphorylation site for cAMP-dependent protein kinase (PKA) on an intracellular domain of the channel. Phosphorylation by various protein kinases has been shown to alter the activity of certain ligand- and voltage-gated ion channels. We have examined the role of phosphorylation by the catalytic subunit of PKA in the regulation of GABAA receptor channel function using whole-cell and excised outside-out patch-clamp techniques. Inclusion of the catalytic subunit of PKA in the recording pipettes significantly reduced GABA-evoked whole-cell and single-channel chloride currents. Both heat inactivation of PKA and addition of the specific protein kinase inhibitor peptide prevented the reduction of GABA-evoked currents by PKA. Neither mean channel open time nor channel conductance was affected by PKA. The reduction in GABA receptor current by PKA was primarily due to a reduction in channel opening frequency.
Porter
N M
NM
Department of Neurology, University of Michigan Medical Center, Ann Arbor 48104.
Twyman
R E
RE
Uhler
M D
MD
Macdonald
R L
RL
D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D018118:Chloride Channels; D000242:Cyclic AMP / Q000494:pharmacology*; D004553:Electric Conductivity; D006358:Hot Temperature; D015640:Ion Channel Gating / Q000187:drug effects; D008565:Membrane Proteins / Q000502:physiology; D051379:Mice; D009474:Neurons / Q000502:physiology*; D010455:Peptides / Q000494:pharmacology; D010766:Phosphorylation; D011189:Potassium Chloride / Q000494:pharmacology; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism / Q000494:pharmacology*; D011963:Receptors, GABA-A / Q000502:physiology*; D013116:Spinal Cord / Q000166:cytology* / Q000196:embryology; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D018118:Chloride Channels; D008565:Membrane Proteins; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011963:Receptors, GABA-A; C061479:protein kinase inhibitor peptide; D005680:gamma-Aminobutyric Acid; D011189:Potassium Chloride; D000242:Cyclic AMP; D011494:Protein Kinases