研究目的
To analyze the expression changes of PACAP and its receptors in the retina after optic nerve crush (ONC) in time and space to provide basis for further research.
研究成果
The spatiotemporal expression of PACAP and its high-affinity receptor PAC1R were remarkably changed after ONC, mainly expressed in the ganglion cell layer of the retina, suggesting their upregulation may play a vital role in RGC death after ONC.
研究不足
The study focused on the spatiotemporal expression changes of PACAP and its receptors in a rodent ONC model, which may not fully replicate human glaucoma conditions. Further studies are needed to elucidate the role of PACAP and PAC1R in RGC apoptosis after ONC.
1：Experimental Design and Method Selection:
The study involved western blot analysis, RT-PCR analysis, and immunofluorescence to examine the expression changes of PACAP and its receptors in the retina after ONC.
2：Sample Selection and Data Sources:
Male Sprague-Dawley rats were used, with ONC performed in the left eye and the right eye as a self-control.
3：List of Experimental Equipment and Materials:
Included RIPA lysis buffer, polyvinylidene difluoride filter membranes, primary antibodies for PACAP, PAC1R, VPAC1R, VPAC2R, and Brn3a, among others.
4：Experimental Procedures and Operational Workflow:
Involved ONC surgery, collection of retinas at predetermined time points, protein and RNA extraction, western blotting, RT-PCR, and immunofluorescence staining.
5：Data Analysis Methods:
Optical density from western blot bands was normalized by GAPDH level, and RT-PCR data were analyzed using the 2-ΔΔCq method.
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