Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Ethanol inhibits gap-junctional coupling between P19 cells.

Kirsten Wentlandt1, Moshe Kushnir, Christian C G Naus

  • 1Toronto Western Research Institute, University Health Network, Toronto, Ontario, Canada.

Alcoholism, Clinical and Experimental Research
|September 15, 2004
PubMed
Summary

This study investigated how ethanol exposure affects gap junctions in P19 cells. Gap junctions are structures that allow direct communication between cells. Researchers found that ethanol exposure significantly reduced the ability of these cells to communicate through gap junctions. This inhibition occurred after both short- and long-term ethanol exposure and did not recover after a 24-hour withdrawal period. The study also found that ethanol selectively reduced levels of a specific protein, Cx43, in the cell membrane. These results suggest that ethanol can disrupt intercellular communication, which may have implications for understanding alcohol's effects on cellular processes.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Clinical implications of frailty in hospitalized patients with pulmonary arterial hypertension.

Chronic respiratory disease·2025
Same author

Cardiorespiratory cross-frequency coupling biomarker for sudden unexpected death in epilepsy.

Epilepsia·2025
Same author

Inhibition of proprotein convertase SKI-1 prevents blood vessel alteration after stroke.

Nature cardiovascular research·2025
Same author

Delta-fast ripple coupling suppression: designing a brain-mimetic stimulation paradigm for seizure abolishment.

Frontiers in neuroscience·2025
Same author

Patterns of Health Care Delivery Among Adults With Heart Failure in the Last Year of Life: A Retrospective Population-Based Study.

Journal of the American Heart Association·2025
Same author

Engaging patients, family caregivers and healthcare providers to develop metrics tailored to a palliative care population: a content validity process.

Journal of patient-reported outcomes·2025

Area of Science:

  • Cell biology and intercellular communication
  • Toxicology and alcohol effects on cellular function
  • Developmental biology and P19 cell models

Background:

Intercellular communication through gap junctions is vital for tissue coordination and development. These junctions enable direct transfer of ions and small molecules between adjacent cells. While their role in physiological processes is well established, the impact of alcohol on gap junction function remains poorly understood. Prior studies have not fully explored how ethanol exposure affects gap junctional communication in cultured cells. This gap motivated researchers to investigate the effects of ethanol on gap junctions in a specific cell line. The P19 cell model is commonly used for studying developmental processes and cellular responses. Ethanol's influence on these cells has not been thoroughly examined in prior literature. This study builds upon existing knowledge of gap junctions by focusing on their function and protein expression after ethanol exposure. No prior work had resolved how long-term ethanol exposure alters gap junctional communication in this model system.

Keywords:
gap junction communicationethanol effects on cellsconnexin 43P19 cell culture

Frequently Asked Questions

Ethanol exposure inhibits gap-junctional communication by reducing dye coupling by more than 50% in P19 cells.

Researchers used the seeding technique with calcein-stained donor cells and measured dye transfer via confocal microscopy and flow cytometry.

The 24-hour withdrawal period tested whether ethanol effects on gap junctions were reversible, but no recovery was observed.

Western blot analysis measured changes in connexin protein levels, specifically Cx26 and Cx43, after ethanol exposure.

Related Experiment Videos

Purpose Of The Study:

This study aimed to determine how ethanol exposure affects gap junctional communication in P19 cells. Researchers wanted to assess both functional and protein-level changes in gap junctions after ethanol treatment. The specific problem addressed is the lack of understanding about how alcohol impacts intercellular communication in cultured cells. The motivation stems from the need to explore ethanol's effects on developmental processes and tissue coordination. The study tested whether ethanol exposure alters dye coupling and connexin protein levels. Researchers also wanted to determine if these effects persist after a withdrawal period. The goal was to provide insights into alcohol's impact on cellular communication mechanisms. This work contributes to understanding how ethanol may disrupt normal cellular interactions.

Main Methods:

The study used P19 cell cultures to investigate ethanol's effects on gap junctions. Researchers applied 20 or 40 mM ethanol for 24 or 48 hours and assessed recovery after a 24-hour withdrawal. Gap-junctional communication was measured using the seeding technique with fluorescent dye. Donor cells stained with calcein were placed on unstained monolayers. Confocal microscopy and flow cytometry quantified dye transfer between cells. Western blot and immunoblot analyses assessed connexin protein levels. Gel electrophoresis separated proteins for further analysis. Antibodies specific to Cx26 and Cx43 were used to detect these proteins. The experimental design allowed researchers to evaluate both functional and molecular changes in gap junctions.

Main Results:

All ethanol treatment regimens reduced dye coupling by more than 50%. This inhibition occurred after 24 and 48 hours of exposure to 20 or 40 mM ethanol. The 24-hour withdrawal period did not restore gap-junctional communication. Exposure to 20 mM ethanol for 48 hours did not change Cx26 protein levels. However, ethanol significantly decreased Cx43 protein levels in cultured P19 cells. This effect was observed in the membrane fraction of the cells. The strongest finding was the selective reduction in Cx43 expression after chronic ethanol exposure. These results suggest that ethanol can inhibit gap junction function in this cell line.

Conclusions:

The authors propose that ethanol exposure inhibits gap-junctional communication in P19 cells. This inhibition was observed after both short- and long-term ethanol exposure. The 24-hour withdrawal period did not reverse the effects of ethanol. Ethanol exposure did not alter Cx26 protein levels in this study. However, Cx43 protein levels were significantly reduced after chronic ethanol exposure. This reduction occurred specifically in the membrane fraction of the cells. The findings suggest that ethanol may selectively affect certain connexin proteins. These results support the hypothesis that alcohol can disrupt intercellular communication mechanisms.

Ethanol did not change Cx26 levels but significantly reduced Cx43 protein levels in the membrane fraction of P19 cells.

The findings suggest ethanol can selectively inhibit gap-junctional communication, potentially disrupting normal cellular coordination.