Desensitization of Nicotine‐Stimulated 86Rb+ Efflux from Mouse Brain Synaptosomes
Abstract
The desensitization of nicotine-stimulated 86Rb+ efflux from synaptosomes prepared from C57BL/6 mouse brain was investigated. Nicotine stimulated a saturable, concentration-dependent efflux of 86Rb+ from synaptosomes (EC50 = 0.60 microM), but the response decreased with time of exposure to nicotine. The rate of decrease of the response (desensitization) increased as the nicotine concentration was increased (EC50 = 0.35 microM; maximal rate of desensitization = 1.1 min-1). Desensitization of nicotine-stimulated 86Rb+ efflux was also observed when synaptosomes were exposed to low (1-200 nM) concentrations of nicotine that caused little or no stimulation of efflux (EC50 = 13 nM). The rate of desensitization observed with low nicotine concentrations (0.30 min-1) was less than that measured at stimulating concentrations. Desensitization was not fully reversible for synaptosomes exposed to nicotine concentrations between 10 nM and 10 microM: Only 60-40% of the control response was regained after a 10-min washout period. The kinetics of functional desensitization were compared with the kinetics of [3H]nicotine binding. [3H]Nicotine binding to midbrain particulate fractions displayed both a fast and a slow phase. The EC50 values for these two phases were 2.6 and 14 nM, respectively. Data obtained from functional desensitization and ligand binding experiments were analyzed using a two-state model. The kinetic constants obtained from the analyses of these two processes were very similar. Overall, the results suggest that nicotinic receptor function measured with ion flux desensitizes when exposed to either stimulating or nonstimulating concentrations of nicotine. In addition, the kinetic properties calculated for the functional desensitization are comparable to those for [3H]nicotine binding.
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In isolated preparations, nicotine can desensitize these receptors at concentrations up to 45-fold lower than those needed to activate them.
“and up to 45 times more potent in vitro (Grady et al., 1994;Marks et al., 1994).”
From Chronic Sazetidine-A at Behaviorally Active Doses Does Not Increase Nicotinic Cholinergic Receptors in Rodent Brain (2012) · cites this paper as Marks et al., 1994
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- Authors
- Marks, M. J., Grady, S. R., Yang, J., Lippiello, P. M., & Collins, A. C.
- Affiliations
- University of Colorado Boulder (US)
- Duke University (US)
- Corresponding author
- Marks, Michael J.
- ORCID
- Marks, Michael J.
- Venue
- Journal of Neurochemistry
- Publisher
- Wiley
- Published
- 1994-12
- Volume
- 63
- Issue
- 6
- Pages
- 2125-2135
- ISSN
- 0022-3042 (ISSN-L)
- 1471-4159
- OpenAlex
- W2137139285
- Cited by
- 61
- References
- 0
- Citations by year
- 2018: 1 · 2015: 2 · 2014: 1 · 2013: 1 · 2012: 3
- FWCI
- 1.01
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Neurochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5k05da000197-07)
- National Institute on Drug Abuse (DA 03194, DA00197, R01DA003194)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nicotinic Acetylcholine Receptors Study
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- nicotine, nicotinic acetylcholine receptors, receptor desensitization, 86Rb efflux, synaptosomes, brain, nicotine binding, two-state model, receptor kinetics, midbrain
- MeSH terms
- Animals, Brain, Drug Tolerance, Female, Kinetics, Mice, Inbred C57BL, Nicotine, Receptors, Nicotinic, Rubidium Radioisotopes, Synaptosomes, Tritium, Mice
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