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Identification of cationic and anionic nicotinic acetylcholine receptor subunits in Helisoma trivolvis
Author(s) -
Zhong Lei,
Wu Yang,
Jiang Chun,
Rehder Vincent
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.933.2
Subject(s) - acetylcholine receptor , lymnaea stagnalis , nicotinic agonist , biology , nicotinic acetylcholine receptor , acetylcholine , protein subunit , microbiology and biotechnology , cholinergic , ion channel , chemistry , receptor , biochemistry , snail , neuroscience , pharmacology , gene , ecology
Many actions of acetylcholine (ACh) are mediated by cation‐selective nicotinic ACh receptors (nAChRs). Our lab found that ACh activates cation‐selective nAChRs to elongate growth cone filopodia on B5 neurons from the pond snail Helisoma trivolvis in vitro . Interestingly, pharmacological evidence revealed the existence of anion‐selective nAChRs in B19 neurons, mediating inhibitory cholinergic responses. Unfortunately, little is known about the molecular nature of these two distinct nAChRs in Helisoma . Using a Helisoma trivolvis transcriptome database, we found putative nAChR partial coding sequences. We cloned two nAChR subunits from Helisoma nervous tissue, HsAChR A and K. HsAChR A was most similar to LnAChR A, a cation‐selective subunit from Lymnaea stagnalis . HsAChR K shared the highest sequence identity with LnAChR K, a predicted anionic receptor subunit. Analysis of the full length sequences showed the putative ACh binding sites, transmembrane domains, and residue motifs for ion selectivity. Single‐cell PCR results revealed the differential expression of HsAChR A and K in B5 and B19 neuron, which might account for their distinct electrophysiological responses to ACh. Identification of the cationic and anionic nAChR subunits is a critical step towards genetic manipulation of this receptor and eventually understanding its roles in brain function and controlling behavior.