Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET

User menu

  • My alerts
  • Log in
  • My Cart

Search

  • Advanced search
Molecular Pharmacology
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET
  • My alerts
  • Log in
  • My Cart
Molecular Pharmacology

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Special Sections
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Submit
  • Visit molpharm on Facebook
  • Follow molpharm on Twitter
  • Follow molpharm on LinkedIn
Research ArticleArticle

Optical Recording Reveals Novel Properties of GSK1016790A-Induced Vanilloid Transient Receptor Potential Channel TRPV4 Activity in Primary Human Endothelial Cells

Michelle N. Sullivan, Michael Francis, Natalie L. Pitts, Mark S. Taylor and Scott Earley
Molecular Pharmacology September 2012, 82 (3) 464-472; DOI: https://doi.org/10.1124/mol.112.078584
Michelle N. Sullivan
Vascular Physiology Research Group, Department of Biomedical Sciences (M.N.S., N.L.P., S.E.) and Department of Biology, Colorado State University, Fort Collins, Colorado (N.L.P.); and Department of Physiology, University of South Alabama College of Medicine, Mobile, Alabama (M.F., M.S.T.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael Francis
Vascular Physiology Research Group, Department of Biomedical Sciences (M.N.S., N.L.P., S.E.) and Department of Biology, Colorado State University, Fort Collins, Colorado (N.L.P.); and Department of Physiology, University of South Alabama College of Medicine, Mobile, Alabama (M.F., M.S.T.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Natalie L. Pitts
Vascular Physiology Research Group, Department of Biomedical Sciences (M.N.S., N.L.P., S.E.) and Department of Biology, Colorado State University, Fort Collins, Colorado (N.L.P.); and Department of Physiology, University of South Alabama College of Medicine, Mobile, Alabama (M.F., M.S.T.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mark S. Taylor
Vascular Physiology Research Group, Department of Biomedical Sciences (M.N.S., N.L.P., S.E.) and Department of Biology, Colorado State University, Fort Collins, Colorado (N.L.P.); and Department of Physiology, University of South Alabama College of Medicine, Mobile, Alabama (M.F., M.S.T.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Scott Earley
Vascular Physiology Research Group, Department of Biomedical Sciences (M.N.S., N.L.P., S.E.) and Department of Biology, Colorado State University, Fort Collins, Colorado (N.L.P.); and Department of Physiology, University of South Alabama College of Medicine, Mobile, Alabama (M.F., M.S.T.)
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Abstract

Critical functions of the vascular endothelium are regulated by changes in intracellular [Ca2+]. Endothelial dysfunction is tightly associated with cardiovascular disease, and improved understanding of Ca2+ entry pathways in these cells will have a significant impact on human health. However, much about Ca2+ influx channels in endothelial cells remains unknown because they are difficult to study using conventional patch-clamp electrophysiology. Here we describe a novel, highly efficient method for recording and analyzing Ca2+-permeable channel activity in primary human endothelial cells using a unique combination of total internal reflection fluorescence microscopy (TIRFM), custom software-based detection, and selective pharmacology. Our findings indicate that activity of the vanilloid (V) transient receptor potential (TRP) channel TRPV4 can be rapidly recorded and characterized at the single-channel level using this method, providing novel insight into channel function. Using this method, we show that although TRPV4 protein is evenly distributed throughout the plasma membrane, most channels are silent even during maximal stimulation with the potent TRPV4 agonist N-((1S)-1-{[4-((2S)-2-{[(2,4-dichlorophenyl)sulfonyl]amino}-3-hydroxypropanoyl)-1-piperazinyl]carbonyl}-3-methylbutyl)-1-benzothiophene-2-carboxamide (GSK1016790A). Furthermore, our findings indicate that GSK1016790A acts by recruiting previously inactive channels, rather than through increasing elevation of basal activity.

Footnotes

  • ↵Embedded Image The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.

  • This work was supported by the National Institutes of Health National Heart, Lung, and Blood Institute [Grant R01-HL091905] (to S.E.), [Grant R01-HL085887] (to M.S.T.).

  • Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.

    http://dx.doi.org/10.1124/mol.112.078584.

  • ABBREVIATIONS:

    TRP
    transient receptor potential
    TIRFM
    total internal reflection fluorescence microscopy
    TRPV
    transient receptor potential vanilloid
    GSK1016790A
    N-((1S)-1-{[4-((2S)-2-{[(2,4-dichlorophenyl)sulfonyl]amino}-3-hydroxypropanoyl)-1-piperazinyl]carbonyl}-3-methylbutyl)-1-benzothiophene-2-carboxamide
    ROI
    region of interest
    RT
    reverse transcription
    PCR
    polymerase chain reaction
    CPA
    cyclopiazonic acid
    4α-PDD
    4α-phorbol 12,13-didecanoate
    HC-067047
    2-methyl-1-[3-(4-morpholinyl)propyl]-5-phenyl-N-[3-(trifluoromethyl)phenyl]-1H-pyrrole-3-carboxamide
    11,12-EET
    11,12-epoxyeicosatrienoic acid.

  • Received March 7, 2012.
  • Accepted June 11, 2012.
  • Copyright © 2012 The American Society for Pharmacology and Experimental Therapeutics
View Full Text

MolPharm articles become freely available 12 months after publication, and remain freely available for 5 years. 

Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page. 

 

  • Click here for information on institutional subscriptions.
  • Click here for information on individual ASPET membership.

 

Log in using your username and password

Forgot your user name or password?

Purchase access

You may purchase access to this article. This will require you to create an account if you don't already have one.
PreviousNext
Back to top

In this issue

Molecular Pharmacology: 82 (3)
Molecular Pharmacology
Vol. 82, Issue 3
1 Sep 2012
  • Table of Contents
  • Table of Contents (PDF)
  • About the Cover
  • Index by author
  • Editorial Board (PDF)
  • Front Matter (PDF)
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Molecular Pharmacology 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.
Optical Recording Reveals Novel Properties of GSK1016790A-Induced Vanilloid Transient Receptor Potential Channel TRPV4 Activity in Primary Human Endothelial Cells
(Your Name) has forwarded a page to you from Molecular Pharmacology
(Your Name) thought you would be interested in this article in Molecular Pharmacology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Research ArticleArticle

TRPV Channel Activity in Endothelial Cells

Michelle N. Sullivan, Michael Francis, Natalie L. Pitts, Mark S. Taylor and Scott Earley
Molecular Pharmacology September 1, 2012, 82 (3) 464-472; DOI: https://doi.org/10.1124/mol.112.078584

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero

Share
Research ArticleArticle

TRPV Channel Activity in Endothelial Cells

Michelle N. Sullivan, Michael Francis, Natalie L. Pitts, Mark S. Taylor and Scott Earley
Molecular Pharmacology September 1, 2012, 82 (3) 464-472; DOI: https://doi.org/10.1124/mol.112.078584
Reddit logo Twitter logo Facebook logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Authorship Contributions
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Cysteine151 in Keap1 Drives CDDO-Me Pharmacodynamic Action
  • Allosteric Modulation of Metabotropic Glutamate Receptor 1
  • Mechanism of Selective Action of Paraherquamide A
Show more Articles

Similar Articles

Advertisement
  • Home
  • Alerts
Facebook   Twitter   LinkedIn   RSS

Navigate

  • Current Issue
  • Fast Forward by date
  • Fast Forward by section
  • Latest Articles
  • Archive
  • Search for Articles
  • Feedback
  • ASPET

More Information

  • About Molecular Pharmacology
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Drug Metabolism and Disposition
  • Journal of Pharmacology and Experimental Therapeutics
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-0111 (Online)

Copyright © 2023 by the American Society for Pharmacology and Experimental Therapeutics