Characterization and staging of outer plexiform layer development in human retina and retinal organoids-Reference-Cited by-同舟云学术

Characterization and staging of outer plexiform layer development in human retina and retinal organoids

Published:2021-12-01 Issue:23 Volume:148 Page:
ISSN:0950-1991
Container-title:Development
language:en
Short-container-title:

Author:

Prameela Bharathan Sumitha¹²^ORCID,Ferrario Angela¹,Stepanian Kayla¹,Fernandez G. Esteban²^ORCID,Reid Mark W.¹,Kim Justin S.¹,Hutchens Chloe¹,Harutyunyan Narine¹²,Marks Carolyn³,Thornton Matthew E.⁴,Grubbs Brendan H.⁴,Cobrinik David¹²⁵⁶⁷^ORCID,Aparicio Jennifer G.¹²^ORCID,Nagiel Aaron¹²⁵^ORCID

Affiliation:

1. Department of Surgery, The Vision Center, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA

2. The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA

3. Core Center of Excellence in Nano Imaging, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA

4. Maternal-Fetal Medicine Division, Department of Obstetrics and Gynecology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA

5. Department of Ophthalmology, Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA

6. Department of Biochemistry and Molecular Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA

7. Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA

Abstract

ABSTRACT The development of the first synapse of the visual system between photoreceptors and bipolar cells in the outer plexiform layer (OPL) of the human retina is crucial for visual processing but poorly understood. By studying the maturation state and spatial organization of photoreceptors, depolarizing bipolar cells and horizontal cells in the human fetal retina, we establish a pseudo-temporal staging system for OPL development that we term OPL-Stages 0 to 4. This was validated through quantification of increasingly precise subcellular localization of bassoon to the OPL with each stage (P<0.0001). By applying these OPL staging criteria to human retinal organoids (HROs) derived from human embryonic and induced pluripotent stem cells, we observed comparable maturation from OPL-Stage 0 at day 100 in culture up to OPL-Stage 3 by day 160. Quantification of presynaptic protein localization confirmed progression from OPL-Stage 0 to 3 (P<0.0001). Overall, this study defines stages of human OPL development through mid-gestation and establishes HROs as a model system that recapitulates key aspects of human photoreceptor-bipolar cell synaptogenesis in vitro.

Funder

Research to Prevent Blindness

National Eye Institute

Las Madrinas

Knights Templar Eye Foundation

Donald E. and Delia B. Baxter Foundation

Children's Hospital Los Angeles

National Institutes of Health

New York University

Publisher

The Company of Biologists

Subject