研究目的
Investigating the roles of retinal microglial cells after activation during the pathogenesis of retinitis pigmentosa (RP) and exploring potential therapeutic targets.
研究成果
During the pathogenesis of RP by MNU, microglial cells exhibit a prominent morphology change, and Müller cells can promote activated microglia infiltration via increased secretion of CX3CL1 and upregulation of CX3CR1 expression in retinal microglial cells. This finding highlights a potential therapeutic target aimed at regulating the microglial response.
研究不足
The primary microglial cells were not directly isolated from the MNU-induced RP rat retinal tissue due to current microglia extract techniques from the retinal tissue of living animal models. The exact molecular mechanism underlying the promotion of chemotaxis initiation in retinal microglial cells by Müller cells was not investigated.
1：Experimental Design and Method Selection:
The study used Sprague Dawley rats to induce RP by intraperitoneal injection of N-methyl-N-nitrosourea (MNU). Retinal histology and electroretinography (ERG) recordings were used to evaluate the effects of MNU on the retinas at serial time points.
2：Sample Selection and Data Sources:
Sixty 10-week-old male Sprague Dawley rats were used. Retinal sections were prepared for histological and immunohistochemical analysis.
3：List of Experimental Equipment and Materials:
Equipment included a Leica TCS SP8 microscope for confocal microscopy, ERG recording equipment, and standard laboratory equipment for histological and biochemical analyses. Materials included MNU, primary antibodies (anti-Iba1, anti-glutamine synthetase), and secondary antibodies.
4：Experimental Procedures and Operational Workflow:
Rats were injected with MNU or saline (control). At specified time points, retinal histology and ERG recordings were performed. Immunofluorescence and Western blot analysis were conducted to assess microglial activation and Müller cell interaction.
5：Data Analysis Methods:
Data were analyzed using one-way ANOVA and Tukey's Honestly Significant Difference test. Confocal microscopy images were analyzed for co-localization using Pearson’s correlation coefficient.
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