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Nicotine up-regulates α4β2 nicotinic receptors and ER exit sites via stoichiometry-dependent chaperoning

Srinivasan, Rahul and Pantoja, Rigo and Moss, Fraser J. and Mackey, Elisha D. W. and Son, Cagdas D. and Miwa, Julie and Lester, Henry A. (2011) Nicotine up-regulates α4β2 nicotinic receptors and ER exit sites via stoichiometry-dependent chaperoning. Journal of General Physiology, 137 (1). pp. 59-79. ISSN 0022-1295. PMCID PMC3010053. doi:10.1085/jgp.201010532. https://resolver.caltech.edu/CaltechAUTHORS:20110208-094115987

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Abstract

The up-regulation of α4β2* nicotinic acetylcholine receptors (nAChRs) by chronic nicotine is a cell-delimited process and may be necessary and sufficient for the initial events of nicotine dependence. Clinical literature documents an inverse relationship between a person’s history of tobacco use and his or her susceptibility to Parkinson’s disease; this may also result from up-regulation. This study visualizes and quantifies the subcellular mechanisms involved in nicotine-induced nAChR up-regulation by using transfected fluorescent protein (FP)-tagged α4 nAChR subunits and an FP-tagged Sec24D endoplasmic reticulum (ER) exit site marker. Total internal reflection fluorescence microscopy shows that nicotine (0.1 µM for 48 h) up-regulates α4β2 nAChRs at the plasma membrane (PM), despite increasing the fraction of α4β2 nAChRs that remain in near-PM ER. Pixel-resolved normalized Förster resonance energy transfer microscopy between α4-FP subunits shows that nicotine stabilizes the (α4)_2(β2)_3 stoichiometry before the nAChRs reach the trans-Golgi apparatus. Nicotine also induces the formation of additional ER exit sites (ERES). To aid in the mechanistic analysis of these phenomena, we generated a β2_(enhanced-ER-export) mutant subunit that mimics two regions of the β4 subunit sequence: the presence of an ER export motif and the absence of an ER retention/retrieval motif. The α4β2_(enhanced-ER-export) nAChR resembles nicotine-exposed nAChRs with regard to stoichiometry, intracellular mobility, ERES enhancement, and PM localization. Nicotine produces only small additional PM up-regulation of α4β2_(enhanced-ER-export) receptors. The experimental data are simulated with a model incorporating two mechanisms: (1) nicotine acts as a stabilizing pharmacological chaperone for nascent α4β2 nAChRs in the ER, eventually increasing PM receptors despite a bottleneck(s) in ER export; and (2) removal of the bottleneck (e.g., by expression of the β2_(enhanced-ER-export) subunit) is sufficient to increase PM nAChR numbers, even without nicotine. The data also suggest that pharmacological chaperoning of nAChRs by nicotine can alter the physiology of ER processes.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1085/jgp.201010532 DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3010053/PubMed CentralArticle
ORCID:
AuthorORCID
Moss, Fraser J.0000-0002-8519-6991
Lester, Henry A.0000-0002-5470-5255
Additional Information:© 2010 Srinivasan et al. This article is distributed under the terms of an Attribution– Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/). Submitted: 8 September 2010; Accepted: 9 December 2010. Published December 27, 2010. We thank Chris Richards and Larry Wade for technical assistance, and K. Scott and A. Goldsipe for assistance with MATLAB coding. This work is supported by grants from the National Institutes of Health (NS-11756 and AG-033954); Targacept Inc.; Louis and Janet Fletcher; the Michael J. Fox Foundation (to R. Srinivasan); and the California Tobacco-Related Disease Research Program (18FT-0066 to R. Srinivasan). Edward N. Pugh Jr. served as editor.
Funders:
Funding AgencyGrant Number
NIHNS-11756
NIHAG-033954
Targacept Inc.UNSPECIFIED
Louis and Janet FletcherUNSPECIFIED
Michael J. Fox FoundationUNSPECIFIED
California Tobacco-Related Disease Research Program18FT-0066
Issue or Number:1
PubMed Central ID:PMC3010053
DOI:10.1085/jgp.201010532
Record Number:CaltechAUTHORS:20110208-094115987
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20110208-094115987
Usage Policy:This article is distributed under the terms of an Attribution– Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
ID Code:22068
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:10 Mar 2011 17:21
Last Modified:09 Nov 2021 16:02

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