TY - JOUR
T1 - Ipsilateral and contralateral retinal ganglion cells express distinct genes during decussation at the optic chiasm
AU - Wang, Qing
AU - Marcucci, Florencia
AU - Cerullo, Isadora
AU - Mason, Carol
N1 - Funding Information:
Supported by NIH National Institutes of Health grants R01 EY012736 and EY015290 (CM) and T32 GM07367 and EY013933 (QW) and Fight for Sight (FM).
Funding Information:
Received June 18, 2016; accepted November 8, 2016; First published November 17, 2016. The authors declare no competing financial interests. Author contributions: QW designed experiments, collected and analyzed data, and wrote the paper; FM collected data for the Foxd1 KO experiments and contributed to writing; IC assisted with the in situ expression studies; CM designed research and wrote the paper with QW. Supported by NIH grants R01 EY012736 and EY015290 (CM) and T32 GM07367 and EY013933 (QW) and Fight for Sight (FM). Qing Wang’s present address: Department of Ophthalmology, Jules Stein Eye Institute, University of California at Los Angeles, Los Angeles, CA 90095. Acknowledgments: We thank Takaaki Kuwajima for help with retrograde labeling experiments, Mika Melikyan for mouse breeding and genotyping, Alexandra Rebsam (INSERM, Paris) for advice on gene profiling experiments, Fiona Doetsch and lab for advice on microarray analysis, Paola Arlotta (Harvard) for advice in the early stages of ISH probe design, and Jane Dodd and members of the Mason lab for discussion and comments on the manuscript. Correspondence should be addressed to either of the following: Carol Mason, Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University. 14-509 P&S Bldg., 630 W. 168th St., New York, NY 10032. E-mail: cam4@columbia.edu; or Qing Wang, UCLA Stein Eye Institute, 100 Stein Plaza, Los Angeles, CA 90095. E-mail: qwang@jsei.ucla.edu. DOI:http://dx.doi.org/10.1523/ENEURO.0169-16.2016 Copyright ©2016 Wang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
Publisher Copyright:
© 2016 Wang et al.
PY - 2016/11/17
Y1 - 2016/11/17
N2 - The increasing availability of transcriptomic technologies within the last decade has facilitated high-throughput identification of gene expression differences that define distinct cell types as well as the molecular pathways that drive their specification. The retinal projection neurons, retinal ganglion cells (RGCs), can be categorized into distinct morphological and functional subtypes and by the laterality of their projections. Here, we present a method for purifying the sparse population of ipsilaterally projecting RGCs in mouse retina from their contralaterally projecting counterparts during embryonic development through rapid retrograde labeling followed by fluorescence-activated cell sorting. Through microarray analysis, we uncovered the distinct molecular signatures that define and distinguish ipsilateral and contralateral RGCs during the critical period of axonal outgrowth and decussation, with more than 300 genes differentially expressed within these two cell populations. Among the differentially expressed genes confirmed through in vivo expression validation, several genes that mark “immaturity” are expressed within postmitotic ipsilateral RGCs. Moreover, at least one complementary pair, Igf1 and Igfbp5, is upregulated in contralateral or ipsilateral RGCs, respectively, and may represent signaling pathways that determine ipsilateral versus contralateral RGC identity. Importantly, the cell cycle regulator cyclin D2 is highly expressed in peripheral ventral retina with a dynamic expression pattern that peaks during the period of ipsilateral RGC production. Thus, the molecular signatures of ipsilateral and contralateral RGCs and the mechanisms that regulate their differentiation are more diverse than previously expected.
AB - The increasing availability of transcriptomic technologies within the last decade has facilitated high-throughput identification of gene expression differences that define distinct cell types as well as the molecular pathways that drive their specification. The retinal projection neurons, retinal ganglion cells (RGCs), can be categorized into distinct morphological and functional subtypes and by the laterality of their projections. Here, we present a method for purifying the sparse population of ipsilaterally projecting RGCs in mouse retina from their contralaterally projecting counterparts during embryonic development through rapid retrograde labeling followed by fluorescence-activated cell sorting. Through microarray analysis, we uncovered the distinct molecular signatures that define and distinguish ipsilateral and contralateral RGCs during the critical period of axonal outgrowth and decussation, with more than 300 genes differentially expressed within these two cell populations. Among the differentially expressed genes confirmed through in vivo expression validation, several genes that mark “immaturity” are expressed within postmitotic ipsilateral RGCs. Moreover, at least one complementary pair, Igf1 and Igfbp5, is upregulated in contralateral or ipsilateral RGCs, respectively, and may represent signaling pathways that determine ipsilateral versus contralateral RGC identity. Importantly, the cell cycle regulator cyclin D2 is highly expressed in peripheral ventral retina with a dynamic expression pattern that peaks during the period of ipsilateral RGC production. Thus, the molecular signatures of ipsilateral and contralateral RGCs and the mechanisms that regulate their differentiation are more diverse than previously expected.
KW - Cell specification
KW - Contralateral
KW - Decussation
KW - Ipsilateral
KW - Optic chiasm
KW - Retinal ganglion cell
UR - http://www.scopus.com/inward/record.url?scp=85014736036&partnerID=8YFLogxK
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U2 - 10.1523/ENEURO.0169-16.2016
DO - 10.1523/ENEURO.0169-16.2016
M3 - Article
C2 - 27957530
AN - SCOPUS:85014736036
SN - 2373-2822
VL - 3
JO - eNeuro
JF - eNeuro
IS - 6
M1 - e0169
ER -