Neural Regeneration Research ›› 2025, Vol. 20 ›› Issue (3): 900-912.doi: 10.4103/NRR.NRR-D-23-01557

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Postnatal development of rat retina: a continuous observation and comparison between the organotypic retinal explant model and in vivo development

Baoqi Hu1, 2, #, Rui Wang2, #, Hanyue Zhang3, Xiou Wang1, Sijia Zhou1, Bo Ma1, Yan Luan3, Xin Wang2, Xinlin Chen3, Zhichao Zhang3, *, Qianyan Kang1, *   

  1. 1Department of Ophthalmology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi Province, China; 2Department of Ophthalmology, The First Affiliated Hospital of Northwest University, Xi’an, Shaanxi Province, China; 3Institute of Neurobiology, Xi’an Jiaotong University Health Science Center, Xi’an, Shaanxi Province, China
  • Online:2025-03-15 Published:2024-06-26
  • Contact: Zhichao Zhang, PhD, zhangzhichao@xjtu.edu.cn; Qianyan Kang, PhD, kangqy@mail.xjtu.edu.cn.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, Nos. 81901156 (to ZZ), 82271200 (to ZZ), 82171308 (to XC); the Fundamental Research Funds for the Central Universities, No. xzy012022035 (to ZZ); the Natural Science Foundation of Shaanxi Province, Nos. 2021JM-261 (to QK), 2023-YBSF-303 (to ZZ); and Traditional Chinese Medicine Project of Shaanxi Province, No. 2019-ZZ-JC047 (to QK).

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Abstract: The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures. However, the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies. Thus, we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina. In this study, we showed that postnatal retinal explants undergo normal development, and exhibit a consistent structure and timeline with retinas in vivo. Initially, we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells. We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin, respectively. Ki-67- and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis, and exhibited a high degree of similarity in abundance and distribution between groups. Additionally, we used Ceh-10 homeodomain-containing homolog, glutamate-ammonia ligase (glutamine synthetase), neuronal nuclei, and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells, Müller glia, mature neurons, and microglia, respectively. The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas. Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development. The findings confirm the accuracy and credibility of this model and support its use for long-term, systematic, and continuous observation.

Key words: bipolar cells, differentiation, in vivo, microglia, Müller glia, organotypic retinal explant culture, postnatal retina development, proliferation, retinal progenitor cells