PARIS, an optogenetic method for functionally mapping gap junctions

  1. Ling Wu
  2. Ao Dong
  3. Liting Dong
  4. Shi-Qiang Wang
  5. Yulong Li  Is a corresponding author
  1. Peking University School of Life Sciences, China

Abstract

Cell-cell communication via gap junctions regulates a wide range of physiological processes by enabling the direct intercellular electrical and chemical coupling. However, the in vivo distribution and function of gap junctions remain poorly understood, partly due to the lack of non-invasive tools with both cell-type specificity and high spatiotemporal resolution. Here we developed PARIS (pairing actuators and receivers to optically isolate gap junctions), a new fully genetically encoded tool for measuring the cell-specific gap junctional coupling (GJC). PARIS successfully enabled monitoring of GJC in several cultured cell lines under physiologically relevant conditions and in distinct genetically defined neurons in Drosophila brain, with ~10-sec temporal resolution and sub-cellular spatial resolution. These results demonstrate that PARIS is a robust, highly sensitive tool for mapping functional gap junctions and study their regulation in both health and disease.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files

Article and author information

Author details

  1. Ling Wu

    State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3921-5626
  2. Ao Dong

    State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2821-9528
  3. Liting Dong

    Peking-Tsinghua Center for Life Sciences, Peking University School of Life Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8396-374X
  4. Shi-Qiang Wang

    State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Yulong Li

    State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China
    For correspondence
    yulongli@pku.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9166-9919

Funding

National Natural Science Foundation of China (Projects 31371442)

  • Yulong Li

National Natural Science Foundation of China (Projects 31671118)

  • Yulong Li

National Natural Science Foundation of China (Projects 31630035)

  • Shi-Qiang Wang

Ministry of Science and Technology of the People's Republic of China (Grant 2015CB856402)

  • Yulong Li

Ministry of Science and Technology of the People's Republic of China (Grant 2016YFA0500401)

  • Shi-Qiang Wang

Beijing Brain Initiation (Z181100001518004)

  • Yulong Li

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2019, Wu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 7,909
    views
  • 1,272
    downloads
  • 33
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Ling Wu
  2. Ao Dong
  3. Liting Dong
  4. Shi-Qiang Wang
  5. Yulong Li
(2019)
PARIS, an optogenetic method for functionally mapping gap junctions
eLife 8:e43366.
https://doi.org/10.7554/eLife.43366

Share this article

https://doi.org/10.7554/eLife.43366

Further reading

    1. Neuroscience
    Andrea Sattin, Chiara Nardin ... Tommaso Fellin
    Research Advance

    Two-photon (2P) fluorescence imaging through gradient index (GRIN) lens-based endoscopes is fundamental to investigate the functional properties of neural populations in deep brain circuits. However, GRIN lenses have intrinsic optical aberrations, which severely degrade their imaging performance. GRIN aberrations decrease the signal-to-noise ratio (SNR) and spatial resolution of fluorescence signals, especially in lateral portions of the field-of-view (FOV), leading to restricted FOV and smaller number of recorded neurons. This is especially relevant for GRIN lenses of several millimeters in length, which are needed to reach the deeper regions of the rodent brain. We have previously demonstrated a novel method to enlarge the FOV and improve the spatial resolution of 2P microendoscopes based on GRIN lenses of length <4.1 mm (Antonini et al., 2020). However, previously developed microendoscopes were too short to reach the most ventral regions of the mouse brain. In this study, we combined optical simulations with fabrication of aspherical polymer microlenses through three-dimensional (3D) microprinting to correct for optical aberrations in long (length >6 mm) GRIN lens-based microendoscopes (diameter, 500 µm). Long corrected microendoscopes had improved spatial resolution, enabling imaging in significantly enlarged FOVs. Moreover, using synthetic calcium data we showed that aberration correction enabled detection of cells with higher SNR of fluorescent signals and decreased cross-contamination between neurons. Finally, we applied long corrected microendoscopes to perform large-scale and high-precision recordings of calcium signals in populations of neurons in the olfactory cortex, a brain region laying approximately 5 mm from the brain surface, of awake head-fixed mice. Long corrected microendoscopes are powerful new tools enabling population imaging with unprecedented large FOV and high spatial resolution in the most ventral regions of the mouse brain.

    1. Neuroscience
    Yafen Li, Yixuan Lin ... Antao Chen
    Research Article

    Concurrent verbal working memory task can eliminate the color-word Stroop effect. Previous research, based on specific and limited resources, suggested that the disappearance of the conflict effect was due to the memory information preempting the resources for distractors. However, it remains unclear which particular stage of Stroop conflict processing is influenced by working memory loads. In this study, electroencephalography (EEG) recordings with event-related potential (ERP) analyses, time-frequency analyses, multivariate pattern analyses (MVPAs), and representational similarity analyses (RSAs) were applied to provide an in-depth investigation of the aforementioned issue. Subjects were required to complete the single task (the classical manual color-word Stroop task) and the dual task (the Sternberg working memory task combined with the Stroop task), respectively. Behaviorally, the results indicated that the Stroop effect was eliminated in the dual-task condition. The EEG results showed that the concurrent working memory task did not modulate the P1, N450, and alpha bands. However, it modulated the sustained potential (SP), late theta (740–820 ms), and beta (920–1040 ms) power, showing no difference between congruent and incongruent trials in the dual-task condition but significant difference in the single-task condition. Importantly, the RSA results revealed that the neural activation pattern of the late theta was similar to the response interaction pattern. Together, these findings implied that the concurrent working memory task eliminated the Stroop effect through disrupting stimulus-response mapping.