关键词:  , 机械敏感通道蛋白；Piezo1；视盘筛板区

Abstract:  Objective To investigate the expression profiles of mechanosensitive channel Piezo family members, Piezo1 and Piezo2, in eyes. Design Experimental study. Participants Nine eyes of 12-week-old adult male C57BL/6N mice, 3 eyes of adult male Piezo1td-Tomato mice, and one paraffin section of a human eyeball were used. Methods RNAscope fluorescence in situ hybridization in the frozen sections of the eyes of the C57BL/6N mice was performed to detect the expression of Piezo1 and Piezo2 mRNA. Quantitative real-time polymerase chain reaction (qPCR) of the eyeball tissues of C57BL/6N mice was performed to detect the relative expression of Piezo1 and Piezo2 mRNA. Western blot of the eyeball tissues of C57BL/6N mice was performed to detect the expression of Piezo1 and Piezo2 proteins. Immunofluorescence staining in the frozen sections of the eyes of Piezo1td-Tomato mice was performed to detect the expression of Piezo1-tdTomato protein. Immunofluorescence staining in the paraffin section of the human eyeball was performed to detect the expression of Piezo1-tdTomato protein. Main Outcome Measures Relative fluorescence intensity and cycle threshold (Ct value). Results RNAscope fluorescence in situ hybridization in the frozen sections of the eyes of the C57BL/6N mice showed that Piezo1 mRNA was expressed in the cornea, iris, ciliary body, lens epithelium, sclera, retina, the neural tissues in the glial laminar region of optic nerve head; and Piezo2 mRNA was expressed in the cornea, iris, and sclera tissues. The qPCR of ocular tissues of the C57BL/6N mice showed that the average Ct value of Piezo1 cDNA was lower than 30 in the cornea, iris, lens, retina, the neural tissues in the glial laminar region of optic nerve head, sclera, and extraocular muscle tissues. The mean Ct value of Piezo2 cDNA was higher than 30 in lens and retinal tissues. Western blot of ocular tissues of the C57BL/6N mice showed that Piezo1 protein was expressed in the cornea, iris, lens, retina, and the neural tissues in the glial laminar region of optic nerve head, sclera, and extraocular muscles. Piezo2 protein is expressed in the cornea, iris, and sclera. Immunofluorescence staining of frozen sections of the eyeball of Piezo1td-Tomato mice showed that Piezo1-tdTomato protein was expressed in the neural tissues in the glial laminar region of optic nerve head. Immunofluorescence staining of the human eyeball paraffin section also showed that Piezo1 protein was expressed in the neural tissues in the laminar region of optic nerve head. Conclusions Piezo1 is widely expressed in the murine eye, including the anterior segment, the posterior segment of the eye, and the optic nerve tissue in the glial lamina area. Piezo1 protein was robustly expressed in the neural tissues in the laminar region of optic nerve head of the human eyeball, which may be an important mechanical sensor that receives and responds to the changes in intraocular pressure, cerebrospinal fluid pressure in the optic nerve sheath space, and the pressure difference between them. (Ophthalmol CHN, 2023, 32: 27-32)

Key words: mechanosensitive channel, Piezo1, laminar region of optic nerve head