Functional mapping and Ca2+ regulation of nicotinic acetylcholine receptor channels in rat hippocampal CA1 neurons.
Diverse subtypes of nicotinic acetylcholine receptors (nAChRs), including fast-desensitizing alpha7-containing receptors thought to be Ca2+-permeable, are expressed in the CNS, where they appear to regulate cognitive processing and synaptic plasticity. To understand the physiological role of nAChRs in regulating neuronal excitability, it is important to know the distribution of functional receptors along the surface of neurons, whether they can increase [Ca2+]i, and/or are regulated by Ca2+. We mapped the distribution of receptors on the membrane of rat hippocampal CA1 stratum radiatum interneurons and pyramidal cells in acute slices by recording nAChR-mediated currents elicited by local UV laser-based photolysis of caged carbachol in patch-clamped neurons. The local application (approximately 7 microm patches) allowed mapping of functional nAChRs along the soma and dendritic tree, whereas the fast uncaging minimized the effects of desensitization of alpha7-containing nAChRs and allowed us to measure the kinetics of responses. The alpha7-containing nAChRs were the predominant subtype on interneurons, and were located primarily at perisomatic sites (<70 microm from the soma; in contrast to the more uniform distribution of glutamate receptors); no currents were detectable on pyramidal neurons. The activation of nAChRs increased [Ca2+]i, indicating that these native receptors in acute slices are significantly Ca2+-permeable, consistent with previous observations made with recombinant receptors. In addition, they exhibited strong desensitization, the rate of recovery from which was controlled by [Ca2+]i. Our results demonstrate the strategic location and Ca2+ regulation of alpha7-containing nAChRs, which may contribute to understanding their involvement in hippocampal plasticity.
Khiroug
Leonard
L
Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA.
Giniatullin
Rashid
R
Klein
Rebecca C
RC
Fayuk
Dmitriy
D
Yakel
Jerrel L
JL
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000528:radiation effects; D004305:Dose-Response Relationship, Drug; D005463:Fluoroacetates; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D066298:In Vitro Techniques; D007395:Interneurons / Q000187:drug effects / Q000378:metabolism; D007424:Intracellular Fluid / Q000378:metabolism; D007700:Kinetics; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D009578:Nitrobenzenes; D018408:Patch-Clamp Techniques; D010777:Photochemistry; D011312:Pressure; D017966:Pyramidal Cells / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000378:metabolism*; D014269:Trifluoroacetic Acid / Q000494:pharmacology / Q000528:radiation effects; D014466:Ultraviolet Rays; D064569:alpha7 Nicotinic Acetylcholine Receptor
C579165:Chrna7 protein, rat; D005463:Fluoroacetates; C088047:N-((alpha-carboxy)-2-nitrobenzyl)carbamoylcholine trifluoroacetate; D009578:Nitrobenzenes; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D002217:Carbachol; D014269:Trifluoroacetic Acid; D002118:Calcium