Skip to main content

Umbrella menu

  • SfN.org
  • eNeuro
  • The Journal of Neuroscience
  • Neuronline
  • BrainFacts.org

Main menu

  • HOME
  • CONTENT
    • Early Release
    • Featured
    • Latest Articles
    • Issue Archive
    • Editorials
    • Research Highlights
  • TOPICS
    • Cognition and Behavior
    • Development
    • Disorders of the Nervous System
    • History, Teaching and Public Awareness
    • Integrative Systems
    • Neuronal Excitability
    • Novel Tools and Methods
    • Sensory and Motor Systems
  • ALERTS
  • FOR AUTHORS
  • EDITORIAL BOARD
  • BLOG
  • ABOUT
    • Overview
    • For the Media
    • Privacy Policy
    • Contact Us
    • Feedback
  • SfN.org
  • eNeuro
  • The Journal of Neuroscience
  • Neuronline
  • BrainFacts.org

User menu

  • My alerts

Search

  • Advanced search
eNeuro
  • My alerts

eNeuro

Advanced Search

Submit a Manuscript
  • HOME
  • CONTENT
    • Early Release
    • Featured
    • Latest Articles
    • Issue Archive
    • Editorials
    • Research Highlights
  • TOPICS
    • Cognition and Behavior
    • Development
    • Disorders of the Nervous System
    • History, Teaching and Public Awareness
    • Integrative Systems
    • Neuronal Excitability
    • Novel Tools and Methods
    • Sensory and Motor Systems
  • ALERTS
  • FOR AUTHORS
  • EDITORIAL BOARD
  • BLOG
  • ABOUT
    • Overview
    • For the Media
    • Privacy Policy
    • Contact Us
    • Feedback
PreviousNext
Research ArticleResearch Article: New Research, Disorders of the Nervous System

Upregulation of nAChRs and Changes in Excitability on VTA Dopamine and GABA Neurons Correlates to Changes in Nicotine-Reward-Related Behavior

Austin T. Akers, Skylar Y. Cooper, Zachary J. Baumgard, Gabriella P. Casinelli, Alicia J. Avelar and Brandon J. Henderson
eNeuro 28 September 2020, 7 (5) ENEURO.0189-20.2020; DOI: https://doi.org/10.1523/ENEURO.0189-20.2020
Austin T. Akers
Department of Biomedical Sciences, Marshall University, Joan C Edwards School of Medicine, Huntington, WV 25703
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Skylar Y. Cooper
Department of Biomedical Sciences, Marshall University, Joan C Edwards School of Medicine, Huntington, WV 25703
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Skylar Y. Cooper
Zachary J. Baumgard
Department of Biomedical Sciences, Marshall University, Joan C Edwards School of Medicine, Huntington, WV 25703
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Gabriella P. Casinelli
Department of Biomedical Sciences, Marshall University, Joan C Edwards School of Medicine, Huntington, WV 25703
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alicia J. Avelar
Department of Biomedical Sciences, Marshall University, Joan C Edwards School of Medicine, Huntington, WV 25703
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Brandon J. Henderson
Department of Biomedical Sciences, Marshall University, Joan C Edwards School of Medicine, Huntington, WV 25703
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Brandon J. Henderson
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Presence of α6-GFP overlaps with presence of TH. A1–A3, 10× images of an α6-GFP mouse coronal brain slice (bregma, −3.1) immunostained with anti-GFP/Alexa Fluor 488 and anti-TH/Alexa Fluor 647. B, 20× image of neurons in the LVTA displaying overlap of α6-GFP and TH presence. C, 20× (with 5× digital zoom) images of LVTA. D, 20× Images of SNc neurons. Scale bars: 100 μm (A1–A3), 50 μm (B1–B3, D1–D3), and 10 μm (C1–C3). Figure Contributions: Brandon J. Henderson performed the experiments.

  • Figure 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2.

    α4-mCherryα6-GFP mice reveal multiple subtypes of nAChRs on VTA pDA neurons. A, Schematic of target mouse brain region (bottom) and sample 10× image of a mouse coronal brain slice at approximately bregma −3.1 mm (top, no immunofluorescence was used here). Scale bar: 250 μm. B, Sample images of control and nicotine-treated VTA DA neurons, set to the same intensity scale, from α4-mCherryα6-GFP mice used in a CPP assay. Scale bar: 10 μm. C, Male and female mice were administered intraperitoneal injections of saline or 0.5 mg/kg nicotine in a CPP assay [n = 14 (8 males and 6 females) and 25 (14 males and 11 females) for saline and nicotine, respectively]. D1–D3, RID of α4α6*, α4*, and α6* nAChRs in saline-treated and nicotine-treated mice (from CPP assays). Individual dots represent the RID of individual mice [n = 11 (6 males and 5 females) and 19 (12 males and 7 females) for saline and nicotine, respectively]. For each mouse, 41–71 putative LVTA DA neurons were imaged. E1, Representative image of a putative VTA DA neuron in a brain slice (bregma, −3.1) from an α6-GFP mouse (scale bars: 20 μm). E2, Representative voltage-clamp recording of putative LVTA DA neurons during a 10-s puff of 300 and 500 nm nicotine. Blue bar indicates duration of nicotine puff and red dotted line represents baseline before nicotine puff. All data are mean ± SEM; *p < 0.05, **p < 0.05, ***p < 0.005; unpaired, two-tailed t test. Exact p values are given in Results. Figure Contributions: Austin T. Akers, Zachary J. Baumgard, Skylar Y. Cooper, Gabriella P. Casinelli, Alicia J. Avelar, and Brandon J. Henderson performed the experiments and analyzed the data.

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3.

    Upregulation of α4α6* and α4* nAChRs in VTA pDA neurons correlates with nicotine reward-related behavior. A1, C1, Representative merged images of LVTA DA neurons in a α4-mCherryα6-GFP brain slice. Scale bar, 10 μm. In nicotine-treated mice, changes in nAChR RID was correlated to CPP score for α4α6* nAChRs, α4* nAChRs, and α6* nAChRs for male (A2, A3, A4) and female (B1, B2, B3) mice. Linear fits (red line) with 95% confidence intervals (dotted red lines). In saline-treated mice, changes in nAChR RID was correlated to CPP score for α4α6* nAChRs (C2), α4* nAChRs (C3) and α6* nAChRs (C4). Linear fits (red line) with 95% confidence intervals (dotted red lines) were applied using Graphpad Prism software. Nicotine correlations used 7 male and 8 female α4-mCherryα6-GFP mice and the saline correlations used 5 α4-mCherryα6-GFP mice (3 males and 2 females). For each mouse, 36–71 neurons were imaged.Figure Contributions: Austin T. Akers, Zachary J. Baumgard, and Brandon J. Henderson performed the experiments and analyzed the data.

  • Figure 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4.

    Upregulation of α4* nAChRs on SNr and VTA putative GABAergic neurons may not correlate with nicotine reward-related behavior. A1, B1, and C1, Representative images of neurons in the LVTA (A1), SNr (B1), or dentate gyrus (C) in a α4-mCherryα6-GFP brain slice. Scale bar, 10 μm (A1 and B1), 250 μm (C1), and 25 μm (C2, C3). A2, B2, RID of saline- and nicotine-treated mice from CPP assays for SNr GABA neurons or VTA GABA neurons. In A2 and B2 the n of individual male and female mice are indicated by individual dots. In nicotine-treated mice, changes in nAChR RID was correlated to CPP score for α4* nAChRs in SNr GABA neurons or VTA GABA neurons. In A3, A4, B3, and B4 the n of individual male and female mice are indicated by individual dots. D1, RID/area of saline- and nicotine-treated mice from CPP assays for dentate gyrus (n = 6, 3 male and 3 female). D2, Changes in nAChR RID/area was correlated to CPP score for α4* nAChRs in the dentate gyrus. Linear fits (red line) with 95% confidence intervals (dotted red lines) were applied using Graphpad Prism software. Data is Mean ± SEM; *p = 0.05, **p = 0.01; unpaired, two-tailed t test. Figure Contribution: Austin T. Akers, Zachary J. Baumgard, Skylar Y. Cooper, Alicia J. Avelar, and Brandon J. Henderson performed the experiments and analyzed the data.

  • Figure 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 5.

    Decreases in VTA pDA neuron firing frequency correlates with reward-related behavior. A, Schematic of target neurons within the LVTA (target bregma, −3.1). B1, B2, Representative images of VTA pDA neurons in DIC (B1) and GFP (B2) imaging modes. Scale bar: 20 μm. Mice used in CPP assays (C) were used to measure firing frequency of pDA neurons in the VTA (D, E). D, Representative cell-attached recordings of VTA pDA neuron baseline firing frequency. E, Mean VTA pDA neuron firing frequency in mice treated with saline or nicotine in CPP assays. Data are mean ± SEM, dots represent data from individual mice (n = 8–9 per condition). F, Reward-related behavior (CPP Score) was correlated to baseline firing frequency of LVTA pDA neurons; *p < 0.05, **p < 0.01; unpaired t test. Exact p values are given in text. Figure Contributions: Brandon J. Henderson performed the experiments and analyzed the data.

  • Figure 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 6.

    Increase in VTA pGABA neuron firing frequency correlates with reward-related behavior. A, Schematic of target pGABA neurons within the LVTA (target bregma, −3.1). B1, B2, Representative images of VTA pGABA neurons in DIC (B1) and GFP (B2) imaging modes. Scale bar: 20 μm. C, Representative cell-attached recordings of VTA pGABA neuron baseline firing frequency from mice assigned to saline or 0.5 mg/kg nicotine CPP cohorts. D, Mean VTA pGABA neuron firing frequency in mice treated with saline or nicotine in CPP assays. Data are mean ± SEM E, Reward-related behavior (CPP Score) were correlated to baseline firing frequency of VTA pGABA neurons; **p < 0.01 with; unpaired t test. Exact p values are given in text. Figure Contributions: Brandon J. Henderson performed the experiments and analyzed the data.

Tables

  • Figures
    • View popup
    Table 1

    Stats table

    FigureFigure panelStatistical testConditionsResults
    2 C Two-way ANOVA interactionF(1,35) = 0.016
    p = 0.90
    Two-way ANOVA sex factorF(1,35) = 0.02
    p = 0.89
    Two-way ANOVA drug factorF(1,35) = 23.9
    p = 0.00002
    Two-way ANOVA means comparisonMale mice (CPP, saline vs nicotine)p = 0.0011
    Two-way ANOVA means comparisonFemale mice (CPP, saline vs nicotine)p = 0.0064
    D1 Two-way ANOVA interactionα4α6* nAChR upregulationF(1,23) = 0.45
    p = 0.51
    Two-way ANOVA sex factorα4α6* nAChR upregulationF(1,25) = 0.12
    p = 0.73
    Two-way ANOVA drug factorα4α6* nAChR upregulationF(1,25) = 19.0
    p = 0.0002
    Two-way ANOVA means comparisonMale mice (α4α6* RID, saline vs nicotine)p = 0.019
    Two-way ANOVA means comparisonFemale mice (α4α6* RID, saline vs nicotine)p = 0.0054
    D2 Two-way ANOVA interactionα4* nAChR upregulationF(1,25) = 0.74
    p = 0.40
    Two-way ANOVA sex factorα4* nAChR upregulationF(1,25) = 0.30
    p = 0.59
    Two-way ANOVA drug factorα4* nAChR upregulationF(1,25) = 15.5
    p = 0.0005
    Two-way ANOVA means comparisonMale mice (α4* RID, saline vs nicotine)p = 0.05
    Two-way ANOVA means comparisonFemale mice (α4* RID, saline vs nicotine)p = 0.008
    D3 Two-way ANOVA interactionα6* nAChR upregulationF(1,25) = 0.32
    p = 0.58
    Two-way ANOVA sex factorα6* nAChR upregulationF(1,25) = 0.87
    p = 0.36
    Two-way ANOVA drug factorα6* nAChR upregulationF(1,25) = 0.086
    p = 0.77
    Two-way ANOVA means comparisonMale mice (α6* RID, saline vs nicotine)p = 0.99
    Two-way ANOVA means comparisonFemale mice (α6* RID, saline vs nicotine)p = 0.99
    3 A2 Linear regressionMale mice, CPP score vs VTA pDA α4α6* nAChR upregulationR2 = 0.67
    F(1,5) = 10.1
    p = 0.025
    A3 Linear regressionMale mice, CPP score vs VTA pDA α4* nAChR upregulationR2 = 0.83
    F(1,5) = 24.4
    p = 0.004
    A4 Linear regressionMale mice, CPP score vs VTA pDA α6* nAChR upregulationR2 = 0.74
    F(1,5) = 14.3
    p = 0.013
    B1 Linear regressionFemale mice, CPP score vs VTA pDA α4α6* nAChR upregulationR2 = 0.82
    F(1,6) = 28.2
    p = 0.002
    B2 Linear regressionFemale mice, CPP score vs VTA pDA α4* nAChR upregulationR2 = 0.81
    F(1,6) = 25.6
    p = 0.002
    B3 Linear regressionFemale mice, CPP score vs VTA pDA α6* nAChR upregulationR2 = 0.07
    F(1,6) = 0.47
    p = 0.52
    C2 Linear regressionCPP score vs VTA pDA α4α6* nAChR upregulationR2 = 0.08
    F(1,3) = 0.24
    p = 0.65
    C3 Linear regressionCPP score vs VTA pDA α4* nAChR upregulationR2 = 0.08
    F(1,3) = 0.26
    p = 0.64
    C4 Linear regressionCPP score vs VTA pDA α6* nAChR upregulationR2 < 0.01
    F(1,3) = 0.01
    p = 0.94
    4 A2 Two-way ANOVA interactionα4* nAChR upregulationF(1,27) = 0.99
    p = 0.33
    Two-way ANOVA sex factorα4* nAChR upregulationF(1,27) = 2.86
    p = 0.10
    Two-way ANOVA drug factorα4* nAChR upregulationF(1,27) = 17.1
    p = 0.0003
    Two-way ANOVA means comparisonMale mice (α4* RID, saline vs nicotine)p = 0.0012
    Two-way ANOVA means comparisonFemale mice (α4* RID, saline vs nicotine)p = 0.05
    A3 Linear regressionMale CPP score vs SNr GABA α4* nAChR upregulationR2 = 0.55
    F(1,6) = 7.39
    p = 0.04
    A4 Linear regressionFemale CPP score vs SNr GABA α4* nAChR upregulationR2 = 0.56
    F(1,6) = 6.45
    p = 0.05
    B2 Two-way ANOVA interactionα4* nAChR upregulationF(1,24) = 0.43
    p = 0.52
    Two-way ANOVA sex factorα4* nAChR upregulationF(1,24) = 0.38
    p = 0.55
    Two-way ANOVA drug factorα4* nAChR upregulationF(1,24) = 14.0
    p = 0.001
    Two-way ANOVA means comparisonMale mice (α4* RID, saline vs nicotine)p = 0.04
    Two-way ANOVA means comparisonFemale mice (α4* RID, saline vs nicotine)p = 0.013
    B3 Linear regressionMale CPP score vs SNr GABA α4* nAChR upregulationR2 < 0.001
    F(1,6) < 0.001
    p = 0.99
    B4 Linear regressionFemale CPP score vs SNr GABA α4* nAChR upregulationR2 = 0.075
    F(1,6) = 0.40
    p = 0.55
    D1 Unpaired t testDentate Gyrus α4* nAChR upregulation, saline vs nicotinep = 0.047
    D2 Linear regressionCPP score vs dentate gyrus α4* nAChR upregulationR2 = 0.02
    F(1,4) = 0.09
    p = 0.78
    5 C Unpaired t testCPP score, saline vs nicotinep = 0.026
    E Unpaired t testVTA pDA neuron firing frequency, saline vs nicotinep = 0.0094
    F Linear regressionCPP Score vs VTA pDA neuron firing frequencyR2 = 0.65
    F(1,16) = 11.2
    p = 0.016
    6 D Unpaired t testVTA pGABA neuron firing frequency, saline vs nicotinep = 0.0056
    E Linear regressionCPP Score vs VTA pGABA neuron firing frequencyR2 = 0.77
    F(1,16) = 20.3
    p = 0.004
    • View popup
    Table 2

    Immunofluorescence

    LVTASNc
    Number of TH+768184
    Number of GFP+644136
    % of GFP+ to TH+83.9%73.9%
    • All GFP+ neurons overlapped with TH+ neurons.

Back to top

In this issue

eneuro: 7 (5)
eNeuro
Vol. 7, Issue 5
September/October 2020
  • Table of Contents
  • Index by author
Email

Thank you for sharing this eNeuro article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Upregulation of nAChRs and Changes in Excitability on VTA Dopamine and GABA Neurons Correlates to Changes in Nicotine-Reward-Related Behavior
(Your Name) has forwarded a page to you from eNeuro
(Your Name) thought you would be interested in this article in eNeuro.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Print
View Full Page PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Citation Tools
Upregulation of nAChRs and Changes in Excitability on VTA Dopamine and GABA Neurons Correlates to Changes in Nicotine-Reward-Related Behavior
Austin T. Akers, Skylar Y. Cooper, Zachary J. Baumgard, Gabriella P. Casinelli, Alicia J. Avelar, Brandon J. Henderson
eNeuro 28 September 2020, 7 (5) ENEURO.0189-20.2020; DOI: 10.1523/ENEURO.0189-20.2020

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Respond to this article
Share
Upregulation of nAChRs and Changes in Excitability on VTA Dopamine and GABA Neurons Correlates to Changes in Nicotine-Reward-Related Behavior
Austin T. Akers, Skylar Y. Cooper, Zachary J. Baumgard, Gabriella P. Casinelli, Alicia J. Avelar, Brandon J. Henderson
eNeuro 28 September 2020, 7 (5) ENEURO.0189-20.2020; DOI: 10.1523/ENEURO.0189-20.2020
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Significance Statement
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgments
    • Footnotes
    • References
    • Synthesis
    • Author Response
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Keywords

  • excitability
  • nicotine
  • nicotinic receptor
  • reward
  • upregulation

Responses to this article

Respond to this article

Jump to comment:

No eLetters have been published for this article.

Related Articles

Cited By...

More in this TOC Section

Research Article: New Research

  • Using cortical neuron markers to target cells in the dorsal cochlear nucleus
  • Parvalbumin Interneurons Are Differentially Connected to Principal Cells in Inhibitory Feedback Microcircuits along the Dorsoventral Axis of the Medial Entorhinal Cortex
  • Traumatic brain injury broadly affects GABAergic signaling in dentate gyrus granule cells
Show more Research Article: New Research

Disorders of the Nervous System

  • Using cortical neuron markers to target cells in the dorsal cochlear nucleus
  • Parvalbumin Interneurons Are Differentially Connected to Principal Cells in Inhibitory Feedback Microcircuits along the Dorsoventral Axis of the Medial Entorhinal Cortex
  • Traumatic brain injury broadly affects GABAergic signaling in dentate gyrus granule cells
Show more Disorders of the Nervous System

Subjects

  • Disorders of the Nervous System
  • Home
  • Alerts
  • Visit Society for Neuroscience on Facebook
  • Follow Society for Neuroscience on Twitter
  • Follow Society for Neuroscience on LinkedIn
  • Visit Society for Neuroscience on Youtube
  • Follow our RSS feeds

Content

  • Early Release
  • Current Issue
  • Latest Articles
  • Issue Archive
  • Blog
  • Browse by Topic

Information

  • For Authors
  • For the Media

About

  • About the Journal
  • Editorial Board
  • Privacy Policy
  • Contact
  • Feedback
(eNeuro logo)
(SfN logo)

Copyright © 2021 by the Society for Neuroscience.
eNeuro eISSN: 2373-2822

The ideas and opinions expressed in eNeuro do not necessarily reflect those of SfN or the eNeuro Editorial Board. Publication of an advertisement or other product mention in eNeuro should not be construed as an endorsement of the manufacturer’s claims. SfN does not assume any responsibility for any injury and/or damage to persons or property arising from or related to any use of any material contained in eNeuro.