Application of the Whole-cell Patch Clamp Combined with Single-cellRT-PCR in an NMDA-induced Glaucoma Mouse Model of the G-proteinPathway in Retinal ON-type Bipolar Cells
Objective: To introduce the application of a retinal whole-cell patch clamp combined with a single-cell reverse transcription polymerase chain reaction in retinal single-cell component analysis and to investigate the effects of N-methyl-D-aspartate (NMDA) on G-protein related mRNA expression by ON-type bipolar cells of the retina. Methods: In this experimental study, twenty 3-5 weeks old C57BL/6J mice were randomly and equally divided into two groups. They then received a 3 μl intravitreal injection of 30 nmol/L NMDA for the induced-glaucoma group or 10 nmol/L phosphate-buffered saline (PBS) for the control group.Twenty-four hours later, ON-type bipolar cells were collected by patch clamp electrode, and RT-PCR was used to determine the relative expression of mRNAs for protein kinase C alpha (PKCα), transient receptor potential melastatin 1 (TRPM1), and G-protein alpha subunit (Gαo). The two groups were compared by Mann-Whitney tests. Results: After intravitreal injection of NMDA, the relative mRNA expression in ONtype bipolar cells of PKCα (0.536±0.339), TRPM1 (0.337±0.271), and Gαo (4.36±1.83) were significantly different from the control eyes (U=0, P=0.001; U=0, P=0.002; U=36, P=0.002). Conclusion: NMDAinduced glaucoma model may affect the signal transduction of G protein-coupled channel of retinal ON bipolar cells.
高青,沈雨濛,娄翔峰,罗雪,卢苇,沈吟. 全细胞膜片钳结合单细胞RT-PCR技术在NMDA诱导的小鼠青光眼模型视网膜双极细胞G蛋白通路研究中的应用[J]. 中华眼视光学与视觉科学杂志, 2017, 19(8): 462-467.
Qing Gao,Yumeng Shen,Xiangfeng Lou,Xue Luo,Wei Lu,Yin Shen. Application of the Whole-cell Patch Clamp Combined with Single-cellRT-PCR in an NMDA-induced Glaucoma Mouse Model of the G-proteinPathway in Retinal ON-type Bipolar Cells. Chinese Journal of Optometry Ophthalmology and Visual science, 2017, 19(8): 462-467. DOI: DOI:10.3760/cma.j.issn.1674-845X.2017.08.003
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