Nephronectin-Integrin α8 signaling is required for proper migration of periocular neural crest cells during chick corneal development

  1. Justin Ma
  2. Lian Bi
  3. James Spurlin
  4. Peter Lwigale  Is a corresponding author
  1. Rice University, United States

Abstract

During development, cells aggregate at tissue boundaries to form normal tissue architecture of organs. However, how cells are segregated into tissue precursors remains largely unknown. Cornea development is a perfect example of this process whereby neural crest cells aggregate in the periocular region prior to their migration and differentiation into corneal cells. Our recent RNA-Seq analysis identified upregulation of Nephronectin (Npnt) transcripts during early stages of corneal development where its function has not been investigated. We found that Npnt mRNA and protein are expressed by various ocular tissues including the migratory periocular neural crest (pNC), which also express the integrin alpha 8 (Itga8) receptor. Knockdown of either Npnt or Itga8 attenuated cornea development, whereas overexpression of Npnt resulted in cornea thickening. Moreover, overexpression of Npnt variants lacking RGD binding sites did not affect corneal thickness. Neither the knockdown or augmentation of Npnt caused significant changes in cell proliferation, suggesting that Npnt directs pNC migration into the cornea. In vitro analyses showed that Npnt promotes pNC migration from explanted periocular mesenchyme, which requires Itga8, focal adhesion kinase (FAK) and Rho kinase (ROCK). Combined, these data suggest that Npnt augments cell migration into the presumptive cornea ECM by functioning as a substrate for Itgα8-positive pNC cells.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 2-7.

Article and author information

Author details

  1. Justin Ma

    Department of Biosciences, Rice University, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Lian Bi

    Department of Biosciences, Rice University, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. James Spurlin

    Department of Biosciences, Rice University, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Peter Lwigale

    Department of Biosciences, Rice University, Houston, United States
    For correspondence
    lwigale@rice.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1799-4905

Funding

National Eye Institute (EY031381)

  • Peter Lwigale

National Eye Institute (EY022158)

  • Peter Lwigale

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

Ethics

Animal experimentation: This study was conducted in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. Fertilized chick embryos incubated between 1 to 17 days were handled according to the approved institutional animal care and use committee (IACUC) protocol (#IACUC-20-190) of Rice University.

Reviewing Editor

  1. Carole LaBonne, Northwestern University, United States

Version history

  1. Received: September 29, 2021
  2. Preprint posted: October 13, 2021 (view preprint)
  3. Accepted: March 2, 2022
  4. Accepted Manuscript published: March 3, 2022 (version 1)
  5. Version of Record published: March 11, 2022 (version 2)

Copyright

© 2022, Ma 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.

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  1. Justin Ma
  2. Lian Bi
  3. James Spurlin
  4. Peter Lwigale
(2022)
Nephronectin-Integrin α8 signaling is required for proper migration of periocular neural crest cells during chick corneal development
eLife 11:e74307.
https://doi.org/10.7554/eLife.74307

Share this article

https://doi.org/10.7554/eLife.74307

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