Functional characterization of acid-sensing ion channels in cultured neurons of rat inferior colliculus.
Acid-sensing ion channels (ASICs), which are widely distributed in the mammalian brain, the spinal cord and the peripheral sensory organs, are ligand-gated cation channels activated by extracellular protons. Abundant experimental evidence shows that ASICs play important roles in physiological/pathological conditions, such as sensory transduction, learning/memory, retinal function, seizure and ischemia. In the auditory system, however, there are only a few studies available describing ASICs in hair cells, the spiral ganglion and the vestibular ganglion. In particular, functional ASICs have not been assessed in the central auditory region, although there is evidence to show their transcription in the inferior colliculus (IC). In the present study, we characterized ASIC-like currents in cultured IC neurons of rats with whole-cell patch-clamp techniques. A rapidly decaying inward current was induced by exogenous application of acidic solution in cultured IC neurons with a response threshold around pH 6.9 and a half activation pH value at 5.92. The current was sensitive to amiloride half-maximal inhibition concentration (IC50)=20.4+/-0.4 microM), an ASIC blocker, and its reversal potential was close to the theoretical Na+ equilibrium potential, indicating that the recorded current was mediated by ASICs. Further experiments revealed the presence of homomeric ASIC1a channels in IC neurons: (1) the ASIC-like current was partially carried by Ca2+ as demonstrated with an ion-substitution protocol and Ca2+ imaging; (2) the current was inhibited by the tarantula venom Psalmotoxin (PcTX1), a specific blocker for homomeric ASIC1a channels; (3) the current could be inhibited by extracellular Ca2+ (IC50=2.31 mM) and Pb2+ (10 microM), confirming the presence of ASIC1a subunit. The presence of functional ASIC2a containing channels was revealed by the Zn2+ (300 microM)-induced enhancement of ASIC-like currents and the absence of functional ASIC3 channels was indicated by the insensitivity of ASIC-like currents to salicylate (1 mM), an ASIC3 subunit blocker. Finally, we show that activation of ASICs by a pH drop could induce membrane depolarization and evoke neuronal firing in IC neurons. Our study clearly demonstrates that functional homomeric ASIC1a channels and ASIC2a-containing channels, but not ASIC3 channels, are present in the IC. We suggest that ASICs should be taken into consideration for their possible functional roles in information processing and pathological processes in the central auditory system.
Zhang
M
M
Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, 443 Yellow Mountain Road, Hefei, China.
Gong
N
N
Lu
Y-G
YG
Jia
N-L
NL
Xu
T-L
TL
Chen
L
L
D000143:Acids / Q000378:metabolism; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004232:Diuretics / Q000494:pharmacology; D004594:Electrophysiology; D005110:Extracellular Space / Q000187:drug effects / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D007245:Inferior Colliculi / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010455:Peptides; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D012459:Salicylates / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism / Q000502:physiology; D026941:Sodium Channel Blockers / Q000494:pharmacology; D015222:Sodium Channels / Q000187:drug effects / Q000502:physiology; D013111:Spider Venoms / Q000494:pharmacology
D000143:Acids; D004232:Diuretics; D007473:Ion Channels; C413209:PcTX1 protein, Psalmopoeus cambridgei; D010455:Peptides; D012333:RNA, Messenger; D012459:Salicylates; D026941:Sodium Channel Blockers; D015222:Sodium Channels; D013111:Spider Venoms; D000584:Amiloride; D012964:Sodium; D002118:Calcium