Neural Regeneration Research ›› 2025, Vol. 20 ›› Issue (12): 3635-3648.doi: 10.4103/NRR.NRR-D-23-02083

Previous Articles    

Single-cell RNA sequencing reveals the heterogeneity and interactions of immune cells and Müller glia during zebrafish retina regeneration

Hui Xu1, 2, *, #, Lining Cao1, #, Yuxi Chen2 , Cuiping Zhou2 , Jie Xu2 , Zhuolin Zhang2 , Xiangyu Li2 , Lihua Liu1 , Jianfeng Lu1, *   

  1. 1 Shanghai YangZhi Rehabilitation Hospital (Shanghai Sunshine Rehabilitation Center), Frontier Science Center for Stem Cell Research, School of Life Sciences and Technology, Tongji University, Shanghai, China;  2 Key Lab of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, NMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology Products, Nantong University, Nantong, Jiangsu Province, China
  • Online:2025-12-15 Published:2025-03-17
  • Contact: Hui Xu, PhD, huixu82@126.com or huixu@ntu.edu.cn; Jianfeng Lu, PhD, lu.jianfeng@tongji.edu.cn.
  • Supported by:
    The study was supported by the National Natural Science Foundation of China, Nos. 81970820 (to HX), 31771644 (to JL), 31930068 (to JL), 82371176 (to JL), 81801331 (to LC); National Key Research and Development Project of China. Nos. 2017YFA0104100 (to JL), 2017YFA0701304 (to HX); Shanghai Yangzhi Rehabilitation Hospital (Shanghai Sunshine Rehabilitation Center) Talent Introduction Plan, No. KYPT202204 (to LC); and the Fundamental Research Funds for the Central Universities, No. 22120230292 (to JL).

PDF

13

Abstract: Inflammation plays a crucial role in the regeneration of fish and avian retinas. However, how inflammation regulates Müller glia (MG) reprogramming remains unclear. Here, we used single-cell RNA sequencing to investigate the cell heterogeneity and interactions of MG and immune cells in the regenerating zebrafish retina. We first showed that two types of quiescent MG (resting MG1 and MG2) reside in the uninjured retina. Following retinal injury, resting MG1 transitioned into an activated state expressing known reprogramming genes, while resting MG2 gave rise to rod progenitors. We further showed that retinal microglia can be categorized into three subtypes (microglia-1, microglia-2, and proliferative) and pseudotime analysis demonstrated dynamic changes in microglial status following retinal injury. Analysis of cell–cell interactions indicated extensive crosstalk between immune cells and MG, with many interactions shared among different immune cell types. Finally, we showed that inflammation activated Jak1–Stat3 signaling in MG, promoting their transition from a resting to an activated state. Our study reveals the cell heterogeneity and crosstalk of immune cells and MG in zebrafish retinal repair, and may provide valuable insights into future mammalian retina regeneration.

Key words: immune cells, inflammation, Jak1–Stat3 signaling, microglia, Müller glia, regeneration, reprogramming, retina, single-cell RNAseq, zebrafish