Brain Region Specific Single-Molecule Fluorescence Imaging

Xu Fu; Faruk H. Moonschi; Ashley M. Fox-Loe; Aaron A. Snell; Deann M. Hopkins; Alicia J. Avelar; Brandon J. Henderson; James R. Pauly; Christopher I. Richards

doi:10.1021/acs.analchem.9b02133

Brain Region Specific Single-Molecule Fluorescence Imaging

Xu Fu, Faruk H. Moonschi, Ashley M. Fox-Loe, Aaron A. Snell, Deann M. Hopkins, Alicia J. Avelar, Brandon J. Henderson, James R. Pauly, Christopher I. Richards

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

We developed an approach utilizing nanoscale vesicles extracted from brain regions combined with single molecule imaging to monitor how an animal's physiological condition regulates the dynamics of protein distributions in different brain regions. This method was used to determine the effect of nicotine on the distribution of receptor stoichiometry in different mouse brain regions. Nicotine-induced upregulation of α4β2 nicotinic acetylcholine receptors (nAChRs) is associated with changes in their expression, trafficking, and stoichiometry. The structural assembly of nAChRs has been quantified in cell culture based systems using single molecule techniques. However, these methods are not capable of quantifying biomolecule assembly that takes place in a live animal. Both nicotine-induced upregulation and changes in nAChR stoichiometry differ across brain regions. Our single molecule approach revealed that nicotine acts differentially across brain regions to alter assembly in response to exposure and withdrawal.

Original language	English
Pages (from-to)	10125-10131
Number of pages	7
Journal	Analytical Chemistry
Volume	91
Issue number	15
DOIs	https://doi.org/10.1021/acs.analchem.9b02133
State	Published - Aug 6 2019

Bibliographical note

Funding Information:
We would like to acknowledge the UKY Light microscopy core for the use of their facilities. Support for this work was provided by the NIH (DA038817, DA040047, and DA046335).

Publisher Copyright:
© 2019 American Chemical Society.

ASJC Scopus subject areas

Analytical Chemistry

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title = "Brain Region Specific Single-Molecule Fluorescence Imaging",

abstract = "We developed an approach utilizing nanoscale vesicles extracted from brain regions combined with single molecule imaging to monitor how an animal's physiological condition regulates the dynamics of protein distributions in different brain regions. This method was used to determine the effect of nicotine on the distribution of receptor stoichiometry in different mouse brain regions. Nicotine-induced upregulation of α4β2 nicotinic acetylcholine receptors (nAChRs) is associated with changes in their expression, trafficking, and stoichiometry. The structural assembly of nAChRs has been quantified in cell culture based systems using single molecule techniques. However, these methods are not capable of quantifying biomolecule assembly that takes place in a live animal. Both nicotine-induced upregulation and changes in nAChR stoichiometry differ across brain regions. Our single molecule approach revealed that nicotine acts differentially across brain regions to alter assembly in response to exposure and withdrawal.",

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note = "Funding Information: We would like to acknowledge the UKY Light microscopy core for the use of their facilities. Support for this work was provided by the NIH (DA038817, DA040047, and DA046335). Publisher Copyright: {\textcopyright} 2019 American Chemical Society.",

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AU - Avelar, Alicia J.

AU - Henderson, Brandon J.

AU - Pauly, James R.

AU - Richards, Christopher I.

N1 - Funding Information: We would like to acknowledge the UKY Light microscopy core for the use of their facilities. Support for this work was provided by the NIH (DA038817, DA040047, and DA046335). Publisher Copyright: © 2019 American Chemical Society.

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AB - We developed an approach utilizing nanoscale vesicles extracted from brain regions combined with single molecule imaging to monitor how an animal's physiological condition regulates the dynamics of protein distributions in different brain regions. This method was used to determine the effect of nicotine on the distribution of receptor stoichiometry in different mouse brain regions. Nicotine-induced upregulation of α4β2 nicotinic acetylcholine receptors (nAChRs) is associated with changes in their expression, trafficking, and stoichiometry. The structural assembly of nAChRs has been quantified in cell culture based systems using single molecule techniques. However, these methods are not capable of quantifying biomolecule assembly that takes place in a live animal. Both nicotine-induced upregulation and changes in nAChR stoichiometry differ across brain regions. Our single molecule approach revealed that nicotine acts differentially across brain regions to alter assembly in response to exposure and withdrawal.

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Brain Region Specific Single-Molecule Fluorescence Imaging

Abstract

Bibliographical note

ASJC Scopus subject areas

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