Absence of Connexin 43 results in smaller retinas and arrested, depolarized retinal progenitor cells in human retinal organoids

Lin Cheng; Matthew R Cring; David A Wadkins; Markus H Kuehn

doi:10.1093/stmcls/sxac017

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Absence of Connexin 43 results in smaller retinas and arrested, depolarized retinal progenitor cells in human retinal organoids

Journal article

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Absence of Connexin 43 results in smaller retinas and arrested, depolarized retinal progenitor cells in human retinal organoids

Lin Cheng, Matthew R Cring, David A Wadkins and Markus H Kuehn

Stem cells (Dayton, Ohio), Vol.40(6), pp.592-604

03/09/2022

DOI: 10.1093/stmcls/sxac017

PMID: 35263762

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Abstract

The development of the vertebrate retina relies on complex regulatory mechanisms to achieve its characteristic layered morphology containing multiple neuronal cell types. While connexin 43 (CX43) is not expressed by mature retinal neurons mutations in its gene GJA1 are associated with microphthalmia and low vision in patients. To delineate how lack of CX43 affects retinal development, GJA1 was disrupted in human induced pluripotent stem cells (hiPSCs) (GJA1-/-) using CRISPR/Cas9 editing, and these were subsequently differentiated into retinal organoids. GJA1-/- hiPSCs do not display defects in self-renewal and pluripotency, but the resulting organoids are smaller with a thinner neural retina and decreased abundance of many retinal cell types. CX43-deficient organoids express lower levels of the neural marker PAX6 and the retinal progenitor cell (RPC) markers PAX6, SIX3, and SIX6. Conversely, expression of the early neuroectoderm markers SOX1 and SOX2 remains high in GJA1-/- organoids throughout their development. Lack of CX43 results in an increased population of CHX10-positive RPCs that are smaller, disorganized, do not become polarized, and possess a limited ability to commit to retinal fate specification. Our data indicate that lack of CX43 causes a developmental arrest in RPCs that subsequently leads to pan-retinal defects and stunted ocular growth.

specification

GJA1

CX43

retinal organoids

retinal progenitor cells

oculodentodigital dysplasia syndrome

Details

Title: Subtitle: Absence of Connexin 43 results in smaller retinas and arrested, depolarized retinal progenitor cells in human retinal organoids
Creators: Lin Cheng - Center for the Prevention and Treatment of Visual Loss, Veterans Affairs Medical Center, Iowa City, IA, USA
Matthew R Cring - University of Iowa
David A Wadkins - Center for the Prevention and Treatment of Visual Loss, Veterans Affairs Medical Center, Iowa City, IA, USA
Markus H Kuehn - University of Iowa
Resource Type: Journal article
Publication Details: Stem cells (Dayton, Ohio), Vol.40(6), pp.592-604
DOI: 10.1093/stmcls/sxac017
PMID: 35263762
NLM abbreviation: Stem Cells
eISSN: 1549-4918
Grant note: name: Center Grant, award: I01 RX000123; DOI: 10.13039/100001818, name: Research to Prevent Blindness; name: University Department of Ophthalmology and Visual Sciences; DOI: 10.13039/100000738, name: U.S. Department of Veterans Affairs; name: United States Government
Language: English
Date published: 03/09/2022
Academic Unit: Ophthalmology and Visual Sciences
Record Identifier: 9984227001602771

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