中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2025, Vol. 20 ›› Issue (3): 900-912.doi: 10.4103/NRR.NRR-D-23-01557

• 原著:视神经损伤修复保护与再生 • 上一篇    

大鼠视网膜的生后发育：器官型视网膜外植体模型与体内发育的比较

  

  • 出版日期:2025-03-15 发布日期:2024-06-26

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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摘要：

虽然器官型视网膜外植体培养已经建立了十多年，与体内实验和细胞培养相比具有一系列独特的优势，但由于缺乏对体内视网膜发育和器官型视网膜外植体培养进行系统和持续的比较，使得该模型在出生后视网膜发育研究中存在争议。因此，实验旨在通过与体内视网膜的比较，验证将该模型用于出生后视网膜发育研究的可行性。实验发现，出生后视网膜外植体可以正常发育，其结构和时间轴与体内视网膜一致。首先，实验利用 SOX2 和 PAX6 免疫染色来识别视网膜祖细胞。然后，分别用 Ki-67和 DCX 免疫染色法检测细胞的增殖和迁移。这些细胞在体内和外植体中都显示出在出生后视网膜形成过程中同步衰退，在丰度和分布上表现出高度的相似性。此外，实验还采用 Chx10，Gl Syn，NeuN 和 Iba1 免疫染色法分别检测了双极细胞、Müller胶质细胞、成熟神经元和小胶质细胞。这些细胞类型出现的时间和空间模式在体内和外植体中表现出显著的一致性。这些结果表明，器官型视网膜外植体培养模型与体内出生后早期视网膜的发育过程高度一致，可以进行长期、系统和连续的观察。此研究提高了该模型在的出生后视网膜发育研究中应用的准确性和可信度。

https://orcid.org/0000-0001-7990-101X (Zhichao Zhang); https://orcid.org/0000-0002-6744-658X (Qianyan Kang)

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