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D Li, QJ Zhang; Proteomic Study on the Retinas of RD, RDS and C3B Mice as Well as Reconfirming of the Differentially Expressed Proteins by Using RT-PCT . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3629.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To analyze the proteomic differential expression in the retinas of rd, rds mice as compared with that of normal C3B mice. Methods: Retinal proteins were collected from rd, rds mice and C3B mice respectively at P7 (postnatal day 7), P12, P21 and P37. The proteins were separated by using 2-DE. Differentially expressed proteins were excised from the gels and in-gel digested by trypsin, and then subjected to peptide analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF). The corresponding mRNA for those differentially expressed proteins were analyzed by using RT-PCR. Results: A lot of differentially expressed proteins were found between rd, rds and C3B mice in 2-DE. Four protein spots were indicated as MGC, PSD95, GRB14 and HPCAL1/NECD. All the 5 genes showed transcriptional changes through RT-PCR analysis. MGC’s protein increased at P12 and then decreased. MGC mRNA in rds retina was higher at P7, P12 and P21 than that in C3B, but was lower at P37. While MGC mRNA in rd was the same as that in C3B at all four ages. PSD95’s protein increased along its development. PSD95 mRNA in rds mice was higher at P7 and P12 than that in C3B, but was lower at P21 and P37. PSD95 mRNA in rd was the same as that in C3B at all four ages. The changes of MGC and PSD95 mRNA at P37 in rds coincided with its 2-DE result. We also have identified one protein as HPCAL1 or NECD. The RT-PCR results of HPCAL1 support its this protein is HPCAL1 but not NECD. The changes of GRB14 mRNA in rds don’t coincide with the proteinaceous changes in 2-DE. Conclusion: MGC, PSD95 and HPCAL1 were found to be expressed differentially, which implys that they may involve in the course of RP.
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