单位：[1]Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, TX 75390[2]McDermott Center for Human Growth and Development, University of Texas Southwestern Medical Center, Dallas, TX 75390[3]Department of Bioinformatics, University of Texas Southwestern Medical Center, Dallas, TX 75390[4]Department of Population and Data Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390[5]Center for Hypothalamic Research and Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390[6]Center for the Genetics of Host Defense, University of Texas Southwestern Medical Center, Dallas, TX 75390[7]Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China 430030.华中科技大学同济医学院附属同济医院眼科[8]Department of Ophthalmology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China 430030.[9]Department of Ophthalmology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China 530021.[10]Shenzhen Key Laboratory of Ophthalmology, Shenzhen Eye Hospital, Shenzhen University School of Medicine, Shenzhen, Guangdong, China 518040.
Retinal disease and loss of vision can result from any disruption of the complex pathways controlling retinal development and homeostasis. Forward genetics provides an excellent tool to find, in an unbiased manner, genes that are essential to these processes. Using NethylNnitrosourea mutagenesis in mice in combination with a screening protocol using optical coherence tomography (OCT) and automated meiotic mapping, we identified 11 mutations presumably causative of retinal phenotypes in genes previously known to be essential for retinal integrity. In addition, we found multiple statistically significant genephenotype associations that have not been reported previously and decided to target one of these genes, Sfxn3 (encoding sideroflexin-3), using CRISPR/Cas9 technology. We demonstrate, using OCT, light microscopy, and electroretinography, that two Sfxn3(-/)mouse lines developed progressive and severe outer retinal degeneration. Electron microscopy showed thinning of the retinal pigment epithelium and disruption of the external limiting membrane. Using singlecell RNA sequencing of retinal cells isolated from C57BL/6J mice, we demonstrate that Sfxn3 is expressed in several bipolar cell subtypes, retinal ganglion cells, and some amacrine cell subtypes but not significantly in M?ller cells or photoreceptors. In situ hybridization confirmed these findings. Furthermore, pathway analysis suggests that Sfxn3 may be associated with synaptic homeostasis. Importantly, electron microscopy analysis showed disruption of synapses and synaptic ribbons in the outer plexiform layer of Sfxn3/ mice. Our work describes a previously unknown requirement for Sfxn3 in retinal function.
基金：
NIHUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [1R01EY022652, U19AI100627, R01AI125581]; National Eye Institute Visual Science Core Grant [EY020799]; Research to Prevent BlindnessResearch to Prevent Blindness (RPB); Patricia and Col. William Massad Retina Research Fund; David M. Crowley Foundation
第一作者单位：[1]Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, TX 75390[7]Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China 430030.
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推荐引用方式(GB/T 7714)：
Bo Chen,Bogale Aredo,Yi Ding,et al.Forward genetic analysis using OCT screening identifies Sfxn3 mutations leading to progressive outer retinal degeneration in mice[J].PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA.2020,117(23):12931-12942.doi:10.1073/pnas.1921224117/-/DCSupplemental.
APA：
Bo Chen,Bogale Aredo,Yi Ding,Xin Zhong,Yuanfei Zhu...&Rafael L. Ufret-Vincenty.(2020).Forward genetic analysis using OCT screening identifies Sfxn3 mutations leading to progressive outer retinal degeneration in mice.PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,117,(23)
MLA：
Bo Chen,et al."Forward genetic analysis using OCT screening identifies Sfxn3 mutations leading to progressive outer retinal degeneration in mice".PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 117..23(2020):12931-12942
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