Abstract
In cholinergic neurons, high-affinity choline uptake in presynaptic terminals is the rate-limiting step in acetylcholine synthesis. Using information provided by the Caenorhabditis elegans Genome Project, we cloned a cDNA encoding the high-affinity choline transporter from C. elegans (cho-1). We subsequently used this clone to isolate the corresponding cDNA from rat (CHT1). CHT1 is not homologous to neurotransmitter transporters, but is homologous to members of the Na+-dependent glucose transporter family. Expression of CHT1 mRNA is restricted to cholinergic neurons. The characteristics of CHT1-mediated choline uptake essentially match those of high-affinity choline uptake in rat brain synaptosomes.
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References
Winkler, J. et al. Essential role of neocortical acetylcholine in spatial memory. Nature 375, 484–487 (1995).
Dutar, P., Bassant, M. H., Senut, M. C. & Lamour, Y. The septohippocampal pathway: structure and function of a central cholinergic system. Physiol. Rev. 75, 393–427 (1995).
Bierer, L. M. et al. Neurochemical correlates of dementia severity in Alzheimer's disease: relative importance of the cholinergic deficits. J. Neurochem. 64, 749–760 (1995).
Tucek, S. Regulation of acetylcholine synthesis in the brain. J. Neurochem. 44, 11–24 (1985).
Haga, T. Synthesis and release of [14C]acetylcholine in synaptosomes. J. Neurochem. 18, 781–798 (1971).
Yamamura, H. I. & Snyder, S. H. Choline: high-affinity uptake by rat brain synaptosomes. Science 178, 626–628 (1972).
Haga, T. & Noda, H. Choline uptake systems of rat brain synaptosomes. Biochim. Biophys. Acta 291, 564–575 (1973).
Kuhar, M. J. & Murrin, L. C. Sodium-dependent, high-affinity choline uptake. J. Neurochem. 30, 15–21 (1978).
Kuhar, M. J., Sethy, V. H., Roth, R. H. & Aghajanian, G. K. Choline: selective accumulation by central cholinergic neurons. J. Neurochem. 20, 581–593 (1973).
Happe, H. K. & Murrin, L. C. High-affinity choline transport sites: use of [3H]hemicholinium-3 as a quantitative marker. J. Neurochem. 60, 1191–1201 (1993).
Simon, J. R. & Kuhar, M. J. Impulse-flow regulation of high affinity choline uptake in brain cholinergic nerve terminals. Nature 255, 162–163 (1975).
Murrin, L. C. & Kuhar, M. J. Activation of high-affinity choline uptake in vitro by depolarizing agents. Mol. Pharmacol. 12, 1082–1090 (1976).
Pascual, J. et al. High-affinity choline uptake carrier in Alzheimer's disease: implications for the cholinergic hypothesis of dementia. Brain Res. 552, 170–174 (1991).
Bissette, G., Seidler, F. J., Nemeroff, C. B. & Slotkin, T. A. High affinity choline transporter status in Alzheimer's disease tissue from rapid autopsy. Ann. NY Acad. Sci. 777, 197–204 (1996).
Nelson, N. The family of Na+/Cl− neurotransmitter transporters. J. Neurochem. 71, 1785–1803 (1998).
Berrard, S. et al. cDNA cloning and complete sequence of porcine choline acetyltransferase: in vitro translation of the corresponding RNA yields an active protein. Proc. Natl. Acad. Sci. USA 84, 9280–9284 (1987).
Alfonso, A. et al. The Caenorhabditis elegans unc-17 gene: a putative vesicular acetylcholine transporter. Science 261, 617–619 (1993).
Wurtman, R. J. Choline metabolism as a basis for the selective vulnerability of cholinergic neurons. Trends Neurosci. 15, 117–122 (1992).
The C. elegans Sequencing Consortium. Genome sequence of the nematode C. elegans: a platform for investigating biology. Science 282, 2012–2018 (1998).
Hediger, M. A. & Rhoads, D. B. Molecular physiology of sodium-glucose cotransporters. Physiol. Rev. 74, 993–1026 (1994).
Nikawa, J., Hosaka, K., Tsukagoshi, Y & Yamashita, S. Primary structure of the yeast choline transport gene and regulation of its expression. J. Biol. Chem. 265, 15996–16003 (1990).
Schloss, P., Mayser, W. & Betz, H. The putative rat choline transporter CHOT1 transports creatine and is highly expressed in neural and muscle-rich tissues. Biochem. Biophys. Res. Commun. 198, 637–645 (1994).
Oh, J. D. et al. Cholinergic neurons in the rat central nervous system demonstrated by in situ hybridization of choline acetyltransferase mRNA. Neuroscience 47, 807–822 (1992).
Roghani, A. et al. Molecular cloning of a putative vesicular transporter for acetylcholine. Proc. Natl. Acad. Sci. USA 91, 10620–10624 (1994).
Vickroy, T. W., Roeske, W. R. & Yamamura, H. I. Sodium-dependent high-affinity binding of [3H]hemicholinium-3 in the rat brain: a potentially selective marker for presynaptic cholinergic sites. Life Sci. 35, 2335–2343 (1984).
Sandberg, K. & Coyle, J. T. Characterization of [3H]hemicholinium-3 binding associated with neuronal choline uptake sites in rat brain membranes. Brain Res. 348, 321–330 (1985).
Kuhar, M. J. & Zarbin, M. A. Synaptosomal transport: a chloride dependence for choline, GABA, glycine and several other compounds. J. Neurochem. 31, 251–256 (1978).
Erickson, J. D. et al. Functional identification of a vesicular acetylcholine transporter and its expression from a “cholinergic” gene locus. J. Biol. Chem. 269, 21929–21932 (1994).
Bejanin, S., Cervini, R., Mallet, J. & Berrard, S. A unique gene organization for two cholinergic markers, choline acetyltransferase and a putative vesicular transporter of acetylcholine. J. Biol. Chem. 269, 21944–21947 (1994).
Barker, L. A. & Mittag, T. W. Comparative studies of substrates and inhibitors of choline transport and choline acetyltransferase. J. Pharmacol. Exp. Ther. 192, 86–94 (1975).
Vogelsberg, V., Neff, N. H. & Hadjiconstantinou, M. Cyclic AMP-mediated enhancement of high-affinity choline transport and acetylcholine synthesis in brain. J. Neurochem. 68, 1062–1070 (1997).
Beeri, R. et al. Enhanced hemicholinium binding and attenuated dendrite branching in cognitively impaired acetylcholinesterase-transgenic mice. J. Neurochem. 69, 2441–2451 (1997).
Kar, S. et al. Amyloid β-peptide inhibits high-affinity choline uptake and acetylcholine release in rat hippocampal slices. J. Neurochem. 70, 2179–2187 (1998).
Kanai, Y. & Hediger, M. A. Primary structure and functional characterization of a high-affinity glutamate transporter. Nature 360, 467–471 (1992).
Cassata, G. et al. Rapid expression screening of Caenorhabditis elegans homeobox open reading frames using a two-step polymerase chain reaction promoter-gfp reporter construction technique. Gene 212, 127–135 (1998).
Mello, C. C., Kramer, J. M., Stinchcomb, D. & Ambros, V. Efficient gene transfer in C. elegans extrachromosomal maintenance and integration of transforming sequences. EMBO J. 10, 3959–3970 (1991).
Saitou, N. & Nei, M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406–425 (1987).
Acknowledgements
We thank Y. Iino for the C. elegans N2 strain, Y. Kobayashi for help with basal forebrain preparations, A. Fire for pPD104.53, S. Yamashita for yeast choline-transporter cDNA, T. Suzuki and Y. Kirino for Torpedo electric lobe and its cDNA library, Y. Koyama for laterodorsal tegmental nucleus, K. Kameyama for suggestions and D. Saffen for English corrections. This work was supported by grants from Japan Science and Technology Corporation (CREST) and the Ministry of Japan Society for the Promotion of Science (Research for Future Program).
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Okuda, T., Haga, T., Kanai, Y. et al. Identification and characterization of the high-affinity choline transporter. Nat Neurosci 3, 120–125 (2000). https://doi.org/10.1038/72059
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DOI: https://doi.org/10.1038/72059
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