The Comparative Pharmacology and Up-Regulation of Rat Neuronal Nicotinic Receptor Subtype Binding Sites Stably Expressed in Transfected Mammalian Cells
Abstract
We stably transfected human embryonic kidney cells (HEK 293 cells) with genes encoding rat neuronal nicotinic receptor alpha2, alpha3, or alpha4 subunits in combination with the beta2 or beta4 subunit to generate six cell lines that express defined subunit combinations that represent potential subtypes of rat neuronal nicotinic acetylcholine receptors (nAChRs). These cell lines were designated KXalpha2beta2, KXalpha2beta4, KXalpha3beta2, KXalpha3beta4, KXalpha4beta2, and KXalpha4beta4. The Kd values of [3H](+/-)epibatidine ([3H]EB) binding to membranes from these six cell lines ranged from approximately 0.02 to 0.3 nM. The pharmacological profiles of the agonist binding sites of these putative nAChR subtypes were examined in competition studies in which unlabeled nicotinic ligands, including 10 agonists and two antagonists, competed against [3H]EB. Most nicotinic ligands examined had higher affinity for the receptor subtypes containing the beta2 subunit compared with those containing the beta4 subunit. An excellent correlation (r > 0.99) of the binding affinities of the 10 agonists was observed between receptors from KXalpha4beta2 cells and from rat forebrain tissue, in which [3H]EB binding represents predominantly alpha4beta2 nAChRs. More important, the affinities (Ki values) for the two tissues were nearly identical. The densities of the binding sites of all six cell lines were increased after a 5-day exposure to (-)-nicotine or the quaternary amine agonist carbachol. These data indicate that these cell lines expressing nAChR subunit combinations should be useful models for investigating pharmacological properties and regulation of the binding sites of potential nAChR subtypes, as well as for studying the properties of nicotinic compounds.
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Epibatidine is a broad-spectrum nAChR ligand that binds with very high affinity (25-300 pM) to all known heteromeric nAChRs.
“Epibatidine is a broad-spectrum nAChR ligand that binds with very high affinity (25–300 pM) to all known heteromeric nAChRs (Avalos et al, 2002; Houghtling et al, 1995; Xiao and Kellar, 2004).”
From Divergent Functional Effects of Sazetidine-A and Varenicline During Nicotine Withdrawal (2013) · cites this paper as Xiao and Kellar, 2004
Cites
Yung-Chi, C., & Prusoff, W. H. (1973) · Biochemical Pharmacology · doi:10.1016/0006-2952(73)90196-2 · Cited by 12,922 · see citing works
Flores, C. M., Rogers, S. W., Pabreza, L. A., Wolfe, B. B., & Kellar, K. J. (1992) · Molecular Pharmacology · doi:10.1016/s0026-895x(25)08832-7 · Cited by 880 · see citing works
Benwell, M. E. M., Balfour, D. J. K., & Anderson, J. M. (1988) · Journal of Neurochemistry · doi:10.1111/j.1471-4159.1988.tb10600.x · Cited by 551 · see citing works
Nicotine binding and nicotinic receptor subunit RNA after chronic nicotine treatment
Marks, M., Pauly, Jr., Gross, S., Deneris, E., Hermans-Borgmeyer, I., Heinemann, S., & Collins, A. (1992) · The Journal of Neuroscience · doi:10.1523/jneurosci.12-07-02765.1992 · Cited by 551 · see citing works
Nicotinic Cholinergic Receptor Binding Sites in the Brain: Regulation in Vivo
Schwartz, R. D., & Kellar, K. J. (1983) · Science · doi:10.1126/science.6828889 · Cited by 479 · see citing works
Self-citationEffects of chronic nicotine infusion on tolerance development and nicotinic receptors
Marks, M. J., Burch, J. B., & Collins, A. C. (1983) · The Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(25)21831-6 · Cited by 464 · see citing works
Albuquerque, E. X., Alkondon, M., Pereira, E. F. R., Castro, N. G., Schrattenholz, A. S., Barbosa, C. T. F., Bonfante-Cabarcas, R. A., Aracava, Y., Eisenberg, H. M., & Maelicke, A. (1997) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(24)36503-6 · Cited by 436 · see citing works
Boulter, J., Connolly, J. G., Deneris, E. S., Goldman, D. A., Heinemann, S. F., & Patrick, J. W. (1987) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.84.21.7763 · Cited by 424 · see citing works
Chavez-Noriega, L. E., Crona, J. H., Washburn, M. S., Urrutia, A., Elliott, K. J., & Johnson, E. C. (1997) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(24)36376-1 · Cited by 405 · see citing works
Chavez-Noriega, L. E., Crona, J. H., Washburn, M. S., Urrutia, A., Elliott, K. J., & Johnson, E. C. (1997) · PubMed · Cited by 392 · see citing works
Peng, X., Gerzanich, V, Anand, R., Whiting, P. J., & Lindstrom, J. (1994) · Molecular Pharmacology · doi:10.1016/s0026-895x(25)09730-5 · Cited by 391 · see citing works
Increased nicotinic receptors in brains from smokers: membrane binding and autoradiography studies
Perry, D. C., Dávila-Garcı́a, M. I., Stockmeier, C. A., & Kellar, K. J. (1999) · The Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(24)38304-1 · Cited by 362 · see citing works
Self-citationPurification and characterization of a nicotinic acetylcholine receptor from rat brain.
Whiting, P. J., & Lindström, J. (1987) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.84.2.595 · Cited by 301 · see citing works
Conroy, W. G., & Berg, D. K. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.9.4424 · Cited by 297 · see citing works
Duvoisin, R. M., Deneris, E. S., Patrick, J., & Heinemann, S. (1989) · Neuron · doi:10.1016/0896-6273(89)90207-9 · Cited by 291 · see citing works
Xiao, Y., Meyer, E. L., Thompson, J. M., Surin, A. M., Wroblewski, J. T., & Kellar, K. J. (1998) · Molecular Pharmacology · doi:10.1124/mol.54.2.322 · Cited by 247 · see citing works
Self-citationHuman Vascular Endothelial Cells Express Functional Nicotinic Acetylcholine Receptors
Macklin, K. D., Maus, A. D. J., Pereira, E. F. R., Albuquerque, E. X., & Conti‐Fine, B. M. (1998) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(24)37807-3 · Cited by 247 · see citing works
Perry, D. C., Xiao, Y., Nguyen, H. N., Musachio, J. L., Dávila‐García, M. I., & Kellar, K. J. (2002) · Journal of Neurochemistry · doi:10.1046/j.1471-4159.2002.00951.x · Cited by 246 · see citing works
Self-citationDavies, A. R. L., HARDICK, D. J., Blagbrough, I. S., Potter, B. V. L., Wolstenholme, A. J., & Wonnacott, S. (1999) · Neuropharmacology · doi:10.1016/s0028-3908(98)00221-4 · Cited by 246 · see citing works
Human and Rodent Bronchial Epithelial Cells Express Functional Nicotinic Acetylcholine Receptors
Maus, A. D. J., Pereira, E. F. R., Karachunski, P. I., Horton, R. M., Navaneetham, D., Macklin, K., Cortes, W. S., Albuquerque, E. X., & Conti-Fine, B. M. (1998) · Molecular Pharmacology · doi:10.1124/mol.54.5.779 · Cited by 234 · see citing works
Breese, C. R., Adams, C. E., Logel, J., Drebing, C. J., Rollins, Y. D., Barnhart, M. M., Sullivan, B. F., DeMasters, B. K., Freedman, R., & Leonard, S. S. (1997) · The Journal of Comparative Neurology · doi:10.1002/(sici)1096-9861(19971027)387:3 · Cited by 231 · see citing works
Methyllycaconitine: a selective probe for neuronal α‐bungarotoxin binding sites
Ward, J. M., Cockcroft, V. B., Lunt, G. G., Smillie, F., & Wonnacott, S. (1990) · FEBS Letters · doi:10.1016/0014-5793(90)81231-c · Cited by 223 · see citing works
Grando, S. A., Horton, R. M., Pereira, E. F. R., Diethelm-Okita, B. M., George, P. M., Albuquerque, E. X., & Conti‐Fine, B. M. (1995) · Journal of Investigative Dermatology · doi:10.1111/1523-1747.ep12325606 · Cited by 210 · see citing works
Whiting, P. J., Schoepfer, R., Lindström, J., & Priestley, T. (1991) · Molecular Pharmacology · doi:10.1016/s0026-895x(25)15355-8 · Cited by 200 · see citing works
Peng, X.-D., Gerzanich, V., Anand, R., Wang, F., & Lindstrom, J. M. (1997) · Molecular Pharmacology · doi:10.1124/mol.51.5.776 · Cited by 191 · see citing works
Wang, F., Nelson, M. E., Kuryatov, A., Olale, F., Cooper, J., Keyser, K., & Lindstrom, J. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.44.28721 · Cited by 187 · see citing works
Chapter 10 Structure and function of neuronal nicotinic acetylcholine receptors
Lindstrom, J. M., Anand, R., Gerzanich, V., Peng, X.-D., Wang, F., & Wells, G. B. (1996) · Progress in brain research · doi:10.1016/s0079-6123(08)62094-4 · Cited by 179 · see citing works
Gopalakrishnan, M., Monteggia, L. M., Anderson, D. J., Molinari, E. J., Piattoni-Kaplan, M., Donnelly-Roberts, D., Arneric, S. P., & Sullivan, J. P. (1996) · The Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(25)12264-7 · Cited by 178 · see citing works
Pharmacology of Neuronal Nicotinic Acetylcholine Receptor Subtypes
Colquhoun, L. M., & Patrick, J. W. (1997) · Advances in pharmacology · doi:10.1016/s1054-3589(08)60072-1 · Cited by 177 · see citing works
Harvey, S. C., Maddox, F. N., & Luetje, C. W. (1996) · Journal of Neurochemistry · doi:10.1046/j.1471-4159.1996.67051953.x · Cited by 173 · see citing works
Parker, M. J., Beck, A., & Luetje, C. W. (1998) · PubMed · Cited by 165 · see citing works
Neuronal Nicotinic Receptor β2 and β4 Subunits Confer Large Differences in Agonist Binding Affinity
Parker, M. J., Beck, A., & Luetje, C. W. (1998) · Molecular Pharmacology · doi:10.1124/mol.54.6.1132 · Cited by 161 · see citing works
Flores, C. M., Dávila‐García, M. I., Ulrich‐Lai, Y. M., & Kellar, K. J. (1997) · Journal of Neurochemistry · doi:10.1046/j.1471-4159.1997.69052216.x · Cited by 142 · see citing works
Self-citationDifferential Agonist Inhibition Identifies Multiple Epibatidine Binding Sites in Mouse Brain
Marks, M. J., Smith, K. W., & Collins, A. C. (1998) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(24)37384-7 · Cited by 140 · see citing works
Pauly, J. R., Marks, M. J., Robinson, S. F., van de Kamp, J. L., & Collins, A. C. (1996) · The Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(25)20143-4 · Cited by 126 · see citing works
Eaton, J. B., Peng, J., Schroeder, K. M., George, A. A., Fryer, J. D., Krishnan, C., Buhlman, L. M., Kuo, Y.-P., Steinlein, O. K., & Lukas, R. J. (2003) · Molecular Pharmacology · doi:10.1124/mol.64.6.1283 · Cited by 117 · see citing works
Lewis, T., Harkness, P. C., Sivilotti, L. G., Colquhoun, D., & Millar, N. S. (1997) · The Journal of Physiology · doi:10.1111/j.1469-7793.1997.299bb.x · Cited by 107 · see citing works
Sanderson, E. M., Drasdo, A. L., McCrea, K. E., & Wonnacott, S. (1993) · Brain Research · doi:10.1016/0006-8993(93)91104-z · Cited by 106 · see citing works
Fitch, R. W., Xiao, Y., Kellar, K. J., & Daly, J. W. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0630641100 · Cited by 99 · see citing works
Self-citationMandelzys, A., Koninck, P. D., & Cooper, E. A. (1995) · Journal of Neurophysiology · doi:10.1152/jn.1995.74.3.1212 · Cited by 95 · see citing works
Stauderman, K. A., Mahaffy, L. S., AKONG, M., Veliçelebi, G., Chavez-Noriega, L. E., Crona, J. H., Johnson, E. C., Elliott, K. J., Gillespie, A. J., Reid, R. T., et al. (1998) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1016/s0022-3565(24)37270-2 · Cited by 90 · see citing works
Stauderman, K. A., Mahaffy, L. S., Akong, M., Veliçelebi, G., Chavez-Noriega, L. E., Crona, J. H., Johnson, E. C., Elliott, K. J., Gillespie, A., Reid, R. T., et al. (1998) · PubMed · doi:10.7270/q28g8j7q · Cited by 86 · see citing works
Autonomic function in mice lacking α5 neuronal nicotinic acetylcholine receptor subunit
Wang, N., Orr‐Urtreger, A., Chapman, J., Rabinowitz, R., Nachman, R., & Korczyn, A. D. (2002) · The Journal of Physiology · doi:10.1113/jphysiol.2001.013456 · Cited by 79 · see citing works
Yamada, H., Ogura, A., Koizumi, S., Yamaguchi, A., & Moriyama, Y. (1998) · Journal of Neuroscience · doi:10.1523/jneurosci.18-13-04946.1998 · Cited by 70 · see citing works
Wong, E. T., Holstad, S. G., Mennerick, S. J., Hong, S. E., Zorumski, C. F., & Isenberg, K. E. (1995) · Molecular Brain Research · doi:10.1016/0169-328x(94)00189-l · Cited by 68 · see citing works
Meyer, E. L., Xiao, Y., & Kellar, K. J. (2001) · PubMed · Cited by 58 · see citing works
Self-citationMeyer, E. L., Xiao, Y., & Kellar, K. J. (2001) · Molecular Pharmacology · doi:10.1016/s0026-895x(24)12620-x · Cited by 58 · see citing works
Self-citationCarbamoylcholine Homologs: Novel and Potent Agonists at Neuronal Nicotinic Acetylcholine Receptors
Jensen, A. A., Mikkelsen, I., Frølund, B., Bräuner‐Osborne, H., Falch, E., & Krogsgaard‐Larsen, P. (2003) · Molecular Pharmacology · doi:10.1124/mol.64.4.865 · Cited by 55 · see citing works
Dávila‐García, M. I., Musachio, J. L., & Kellar, K. J. (2003) · Journal of Neurochemistry · doi:10.1046/j.1471-4159.2003.01774.x · Cited by 42 · see citing works
Self-citationAvila, A. M., Dávila‐García, M. I., Ascarrunz, V. S., Xiao, Y., & Kellar, K. J. (2003) · Molecular Pharmacology · doi:10.1124/mol.64.4.974 · Cited by 40 · see citing works
Self-citationActivation and Ca2+Permeation of Stably Transfected α3/β4 Neuronal Nicotinic Acetylcholine Receptor
Zhang, J., Xiao, Y., Abdrakhmanova, G., et al. (1999) · Molecular Pharmacology · doi:10.1124/mol.55.6.970 · Cited by 37 · see citing works
Self-citationKellar, K. J., Giblin, B. A., & Lumpkin, M. D. (1989) · Progress in brain research · doi:10.1016/s0079-6123(08)62480-2 · Cited by 29 · see citing works
Self-citationActivation and Ca2+ Permeation of Stably Transfected α3/β4 Neuronal Nicotinic Acetylcholine Receptor
Zhang, J., Xiao, Y., Abdrakhmanova, G., et al. (1999) · Molecular Pharmacology · doi:10.1016/s0026-895x(24)23246-6 · Cited by 1 · see citing works
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Albuquerque, E. X., Pereira, E. F. R., Alkondon, M., & Rogers, S. W. (2009) · Physiological Reviews · doi:10.1152/physrev.00015.2008 · Cited by 1,724 · see citing works
Structural and functional diversity of native brain neuronal nicotinic receptors
Gotti, C., Clementi, F., Fornari, A., Gaimarri, A., Guiducci, S., Manfredi, I., Moretti, M., Pedrazzi, P., Pucci, L., & Zoli, M. (2009) · Biochemical Pharmacology · doi:10.1016/j.bcp.2009.05.024 · Cited by 469 · see citing works
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Xiao, Y., Fan, H., Musachio, J. L., Wei, Z.-L., Chellappan, S. K., Kozikowski, A. P., & Kellar, K. J. (2006) · Molecular Pharmacology · doi:10.1124/mol.106.027318 · Cited by 141 · see citing works
Self-citationMao, D., Perry, D. C., Yasuda, R. P., Wolfe, B. B., & Kellar, K. J. (2008) · Journal of Neurochemistry · doi:10.1111/j.1471-4159.2007.05011.x · Cited by 133 · see citing works
Perry, D. C., Mao, D., Gold, A. B., McIntosh, J. M., Pezzullo, J. C., & Kellar, K. J. (2007) · The Journal of Pharmacology and Experimental Therapeutics · doi:10.1124/jpet.107.121228 · Cited by 106 · see citing works
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Portugal, G. S., Wilkinson, D. S., Turner, J. R., Blendy, J. A., & Gould, T. J. (2012) · Neurobiology of Learning and Memory · doi:10.1016/j.nlm.2012.04.003 · Cited by 79 · see citing works
Tumkosit, P., Kuryatov, A., Luo, J., & Lindstrom, J. (2006) · Molecular Pharmacology · doi:10.1124/mol.106.027326 · Cited by 73 · see citing works
Marks, M. J., McClure-Begley, T. D., Whiteaker, P., Salminen, O., Brown, R. W. B., Cooper, J., Collins, A. C., & Lindstrom, J. M. (2011) · The Journal of Pharmacology and Experimental Therapeutics · doi:10.1124/jpet.110.178236 · Cited by 70 · see citing works
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Marks, M. J., Whiteaker, P., & Collins, A. C. (2006) · Molecular Pharmacology · doi:10.1124/mol.106.025338 · Cited by 53 · see citing works
Lomazzo, E., Hussmann, G. P., Wolfe, B. B., Yasuda, R. P., Perry, D. C., & Kellar, K. J. (2011) · Journal of Neurochemistry · doi:10.1111/j.1471-4159.2011.07408.x · Cited by 37 · see citing works
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- Authors
- Xiao, Y., & Kellar, K. J.
- Affiliations
- Georgetown University (US)
- ORCID
- Kellar, Kenneth J.
- Venue
- The Journal of Pharmacology and Experimental Therapeutics
- Publisher
- Elsevier BV
- Published
- 2004-07
- Volume
- 310
- Issue
- 1
- Pages
- 98-107
- ISSN
- 0022-3565 (ISSN-L)
- OpenAlex
- W2105904453
- Cited by
- 148
- References
- 53
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 4 · 2022: 1 · 2021: 2 · 2020: 3 · 2019: 2 · 2018: 6 · 2017: 4 · 2016: 4 · 2015: 10 · 2014: 12 · 2013: 12 · 2012: 11
- FWCI
- 4.14
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Pharmacology and Experimental Therapeutics (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01da006486-10)
- National Institute on Drug Abuse (DA-06486, DA 12976)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nicotinic Acetylcholine Receptors Study
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Keywords
- neuronal nicotinic acetylcholine receptors, nAChR subtypes, alpha2 subunit, alpha3 subunit, alpha4 subunit, beta2 subunit, beta4 subunit, epibatidine binding, carbachol, receptor subunit composition
- MeSH terms
- Animals, Binding Sites, Carbachol, Cell Line, Humans, Neurons, Nicotine, Pyridines, Receptors, Nicotinic, Transfection, Tritium, Up-Regulation, Gene Expression, Cholinergic Agonists, Nicotinic Agonists, Bridged Bicyclo Compounds, Heterocyclic, Rats
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