Journal
JOURNAL OF NEUROCHEMISTRY
Volume 91, Issue 3, Pages 766-768Publisher
WILEY
DOI: 10.1111/j.1471-4159.2004.02746.x
Keywords
acetylcholine; choline; choline high affinity transporter (CHT); PC12 cells; vesicular acetylcholine transporter; vesamicol
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Funding
- NINDS NIH HHS [NS15047] Funding Source: Medline
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Previously published results appeared to show that vesicular acetylcholine transporter (VAChT) does not transport choline (Ch). Because it is uniquely suited to detect transport of weakly bound substrates, a recently developed assay that detects transmembrane reorientation of the substrate binding site was used to re-examine transport selectivity. Rat VAChT was expressed in PC12(A1237) cells, postnuclear supernatant-containing microvesicles was prepared, and the reorientation assay was conducted with unlabeled Ch and tetramethylammonium (TMA). Also, [C-14]Ch and [H-3]acetylcholine (ACh) were used in an optimized accumulation assay. The results demonstrate that Ch is transported at least as well as ACh is, but with sevenfold lower affinity. Even TMA is transported, but with 26-fold lower affinity. Ch transport by VAChT is of interest in view of the possibilities that Ch (i) occurs at higher concentration than ACh does in terminal cytoplasm under some conditions, and (ii) is an agonist for alpha7 nicotinic receptors.
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