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A subtype of nicotinic cholinergic receptor in rat brain is composed of alpha 4 and beta 2 subunits and is up-regulated by chronic nicotine treatment.
Abstract
The subunit composition and pharmacological regulation of rat neuronal nicotinic cholinergic receptors were assessed. Specific immunoprecipitation was determined in solubilized rat brain homogenates using [3H]cytisine, a high affinity agonist at nicotinic receptors, in conjunction with polyclonal antisera generated against nonhomologous domains of the various subunits comprising this receptor class. In all brain regions tested, only antisera generated against the alpha 4 and beta 2 subunits were able to immunoprecipitate specifically receptors labeled by [3H]cytisine. Thus, these sera were further characterized in order to validate and optimize their use in the immunoprecipitation protocol. Preincubation of solubilized receptors from rat forebrain with antisera generated against the alpha 2, alpha 3, alpha 5, beta 3, or beta 4 subunits did not decrease the amount of precipitable alpha 4 or beta 2 subunit. On the other hand, when either anti-alpha 4 or anti-beta 2 serum was used to immunoprecipitate solubilized receptors from rat forebrain, the supernatants contained little if any remaining receptors that could be specifically precipitated by either antibody. Because these antisera do not cross-react, the data indicate that alpha 4 and beta 2 subunits are associated with each other in at least one neuronal nicotinic receptor subtype that has high affinity for agonists. Moreover, these results imply that all alpha 4 subunits that are labeled by [3H]cystisine are coupled to beta 2 subunits. We also present evidence that the alpha 4/beta 2 subtype characterized in this report is significantly increased in the cortex of rats chronically treated with nicotine.
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“Importantly, nicotine induced increases in nAChR, termed “upregulation”, are conferred by a specific microdomain that is in the β2-subunit 34 and thus this upregulation is confined to nAChR that contain the β2-subunit4, 19, 31, 35-38.”
From β2-Nicotinic Acetylcholine Receptor Availability During Acute and Prolonged Abstinence From Tobacco Smoking (2009) · cites this paper as [19]
In the citing paper
“Immunoprecipitation with subunit-selective antibodies and studies in genetic knockout models demonstrate that chronic administration of nicotine increases predominantly the ␣42 subtype of nAChRs in rodent brain (Flores et al., 1992;Mao et al., 2008;Moretti et al., 2010;Marks et al., 2011)”
From Chronic Sazetidine-A at Behaviorally Active Doses Does Not Increase Nicotinic Cholinergic Receptors in Rodent Brain (2012) · cites this paper as Flores et al., 1992
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Mammalian nicotinic acetylcholine receptors: from structure to function
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
Presynaptic nicotinic ACh receptors
Wonnacott, S. (1997) · Trends in Neurosciences · doi:10.1016/s0166-2236(96)10073-4 · Cited by 1,308
The Diversity of Neuronal Nicotinic Acetylcholine Receptors
Sargent, P. B. (1993) · Annual Review of Neuroscience · doi:10.1146/annurev.ne.16.030193.002155 · Cited by 1,052
Miyamoto, S., Duncan, G. E., Marx, C. E., & Lieberman, J. A. (2005) · Molecular Psychiatry · doi:10.1038/sj.mp.4001556 · Cited by 988
Neuronal nicotinic receptors in the human brain
Paterson, D., & Nordberg, A. K. (2000) · Progress in Neurobiology · doi:10.1016/s0301-0082(99)00045-3 · Cited by 792
Metadata
- Authors
- Flores, C. M., Rogers, S. W., Pabreza, L. A., Wolfe, B. B., & Kellar, K. J.
- Affiliations
- Georgetown University Medical Center (US)
- University of Colorado Denver (US)
- Georgetown University (US)
- ORCID
- Kellar, K J
- Venue
- Molecular Pharmacology
- Publisher
- Elsevier BV
- Published
- 1992-01
- Volume
- 41
- Issue
- 1
- Pages
- 31-37
- ISSN
- 0026-895X (ISSN-L)
- OpenAlex
- W2116249462
- Cited by
- 880
- References
- 0
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 3 · 2023: 3 · 2022: 7 · 2021: 7 · 2020: 7 · 2019: 10 · 2018: 7 · 2017: 9 · 2016: 18 · 2015: 25 · 2014: 25 · 2013: 22 · 2012: 21
- FWCI
- 8.73
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Molecular Pharmacology (publishedVersion)
- PubMed (publishedVersion)
- PubMed
- Funders
- National Institutes of Health (5r01ag009884-05, 5r01da006486-10, 5r01ns011549-24)
- National Institute on Aging (AG 09884)
- National Institute on Drug Abuse (DA-06486)
- National Institute of Neurological Disorders and Stroke (NS11549)
- 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
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Smoking Behavior and Cessation
- Keywords
- nicotinic acetylcholine receptors, receptor subunit composition, chronic nicotine treatment, receptor up-regulation, rat cortex, forebrain, neuronal nicotinic receptors
- MeSH terms
- Quinolizidine Alkaloids, Alkaloids, Animals, Antibodies, Azocines, Brain, Kinetics, Male, Nicotine, Precipitin Tests, Quinolizines, Rats, Inbred Strains, Receptors, Nicotinic, Time Factors, Tritium, Up-Regulation, Macromolecular Substances, Rats
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