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目的:以PPE-GFP转基因小鼠为研究工具,观察绿色荧光蛋白(GFP)阳性的脑啡肽(ENK)能神经元与钙结合蛋白D28K(CB)、钙视网膜蛋白(CR)和小白蛋白(PV)等钙结合蛋白(CaBPs)成员在视网膜的分布及共存情况。方法:利用免疫组织化学和免疫荧光双标染色的方法。结果:GFP阳性的ENK能细胞主要分布在视网膜内核层内缘,少量分布在节细胞层。所有的GFP阳性细胞均与神经元标志物NSE共存,但不与星形胶质细胞标志物GFAP共存。GFP与CB、CR和PV均有部分共存,其中GFP/CB共存神经元占GFP阳性细胞的8.65%,占CB阳性细胞的5.84%;GFP/CR共存神经元占GFP阳性细胞的18.18%,占CR阳性细胞的14.28%,且共存细胞仅见于内核层;GFP/PV共存细胞占GFP阳性细胞的68.75%,占PV阳性细胞的91.67%,共存细胞主要位于内核层,少量见于节细胞层。结论:ENK能神经元在视网膜内具有板层特异性的分布特点和与钙结合蛋白成员有不同的共存模式,上述结果为深入研究小鼠视网膜ENK能神经元的功能意义提供了形态学依据。
OBJECTIVE: To investigate the relationship between enkephalin (ENK) -negative neurons and calcium-binding protein D28K (CB), calcium retinal protein (CR) and small albumin in PPE-GFP transgenic mice. (PV) and other calcium-binding protein (CaBPs) members in the retina of the distribution and coexistence. Methods: Immunohistochemistry and immunofluorescence double-stained staining method. Results: The GFP-positive ENK cells were mainly located in the inner edge of the inner retinal layer, a small amount of distribution in the cell layer. All GFP positive cells coexisted with the neuronal marker NSE but did not coexist with the astrocyte marker GFAP. GFP and CB, CR and PV partially coexisted. GFP / CB co-existing neurons accounted for 8.65% of GFP-positive cells, accounting for 5.84% of CB-positive cells; GFP / CR coexisting neurons accounted for 18.18% of GFP- CR cells were 14.28%, and the coexisting cells were only found in the inner nuclear layer. The number of GFP / PV coexisting cells accounted for 68.75% of the GFP positive cells, accounting for 91.67% of the PV positive cells. The coexisting cells mainly located in the inner nuclear layer and a few in the ganglion cell layer. CONCLUSIONS: ENK neurons have platelet-specific distribution in the retina and different coexistence patterns with calcium-binding protein members. These results provide morphological evidence for further study on the functional significance of ENK neurons in mouse retina.