The reducing agent dithiothreitol (DTT) does not abolish the inhibitory nicotinic response recorded from rat dorsolateral septal neurons.
Previous intracellular recordings have demonstrated that dorsolateral septal nucleus (DLSN) neurons express a novel nicotinic receptor which produces a direct membrane hyperpolarization when activated by nicotinic agonists. Activation of the classical excitatory nicotinic receptors has been shown to require a disulfide bond involving the cysteines at positions 192 and 193 of the alpha subunits of the receptor. Reduction of this cystine bond with dithiothreitol (DTT) abolishes agonist activation of excitatory nicotinic receptors. We have now examined whether DTT treatment of the inhibitory nicotinic receptor on DLSN neurons also abolishes the inhibitory nicotinic response. We find that the inhibitory response persists after treatment of the neurons with 1 mM DTT, even if the reduction is followed by alkylation of the receptor with bromoacetylcholine to prevent possible reformation of disulfide bonds. This result suggests that the agonist binding site on the inhibitory nicotinic receptor does not require an intact disulfide bond, similar to the bond on the alpha subunit of the excitatory nicotinic receptor, for agonist activation of the receptor. Some of these results have been previously reported in abstract form.
Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston 77555-1031.
E M
EM
Sorenson
J P
JP
Gallagher
D000109:Acetylcholine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D003553:Cystine / Q000187:drug effects; D004246:Dimethylphenylpiperazinium Iodide / Q000494:pharmacology; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D010084:Oxidation-Reduction; D010830:Physostigmine / Q000494:pharmacology; D017434:Protein Structure, Tertiary / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D011978:Receptors, Nicotinic / Q000187:drug effects*; D012688:Septum Pellucidum / Q000187:drug effects*; D014726:Vestibular Nuclei / Q000187:drug effects*
D011978:Receptors, Nicotinic; D003553:Cystine; D004246:Dimethylphenylpiperazinium Iodide; D004228:Dithionitrobenzoic Acid; D010830:Physostigmine; D000109:Acetylcholine; D004229:Dithiothreitol