G蛋白偶联受体56基因敲除抑制少突胶质前体细胞成熟

发布时间：2018-05-27 04:34

本文选题：G蛋白偶联受体 + 胼胝体　；参考：《中国病理生理杂志》2014年03期

【摘要】：目的:探讨G蛋白偶联受体56(GPR56)基因敲除对小鼠脑胼胝体内轴突髓鞘化和少突胶质前体细胞(OPCs)成熟的影响。方法:筛选出GPR56基因杂合型(GPR56+/-)和敲除型(GPR56-/-)小鼠36只,分为GPR56+/-和GPR56-/-组,每组18只。每组根据小鼠出生后时间分为出生后7 d(P7)、14 d(P14)、21 d(P21)和28d(P28)4个亚组。应用FluoroMyelin染色观察P14、P21和P28 GPR56+/-和GPR56-/-小鼠脑胼胝体内髓鞘形成。用电镜观察P28 GPR56+/-和GPR56-/-小鼠胼胝体内轴突髓鞘化,比较髓鞘的厚度。用荧光免疫组化染色观察P7GPR56+/-和GPR56-/-小鼠胼胝体内血小板源性生长因子α受体阳性(PDGF-αR+)细胞(即OPCs)的数量。用原位杂交监测P28 GPR56+/-和GPR56-/-小鼠胼胝体内髓鞘蛋白脂质蛋白阳性(PLP+)细胞数。用出生后1 d的GPR56+/-和GPR56-/-小鼠脑皮质做体外OPCs培养并诱导其分化成熟,观察pro-oligodendroblast、immature oligodendrocyte和mature oligodendrocyte阶段O4+细胞百分比。结果:与GPR56+/-小鼠比较,在P14、P21和P28GPR56-/-小鼠脑胼胝体中髓鞘的形成明显减少。电镜见P28 GPR56-/-小鼠脑胼胝体内髓鞘化轴突的数量明显减少,髓鞘g-ratio值变大,髓鞘厚度变薄。荧光免疫组化和原位杂交结果显示P7 GPR56+/-和GPR56-/-小鼠胼胝体内PDGF-aR+细胞数量无差异,但P28 GPR56+/-小鼠胼胝体内PLP+细胞数明显多于P28 GPR56-/-小鼠。体外细胞培养结果显示在pro-oligodendroblast阶段GPR56-/-O4+细胞百分比明显多于GPR56+/-O4+细胞,在immature oligodendrocyte和mature oligodendrocyte阶段GPR56-/-O4+细胞百分比明显少于GPR56+/-O4+细胞。结论:GPR56蛋白可能参与了脑白质轴突髓鞘化和OPCs的成熟。
[Abstract]:Aim: to investigate the effects of G protein-coupled receptor 56 (GPR56) knockout on axonal myelination and oligodendrocyte precursor cell maturation in the corpus callosum of mice. Methods: a total of 36 GPR56 gene heterozygous (GPR56 / -) and knockout GPR56 /-(-) mice were selected and divided into GPR56 / R and GPR56-r- groups with 18 in each group. Each group was divided into 4 subgroups according to the postnatal time: P7, P7, P14, P14, P21, and P28, respectively. FluoroMyelin staining was used to observe the formation of myelin in the corpus callosum of P14 P21 and P28 GPR56 r-and GPR56-r- mice. The myelination of axons in the corpus callosum of P28 GPR56 r- and GPR56-r- mice was observed by electron microscope and the thickness of myelin sheath was compared. The number of platelet-derived growth factor 伪 receptor positive PDGF- 伪 R cells in the corpus callosum of P7GPR56 r- and GPR56-r- mice was observed by fluorescence immunohistochemical staining. In situ hybridization was used to monitor the number of myelin lipid protein positive cells in the corpus callosum of P28 GPR56 r- and GPR56-r- mice. In vitro OPCs culture and differentiation and maturation were conducted in the cerebral cortex of GPR56 / and GPR 56-r- mice on day 1 after birth. The percentage of O4 cells in pro-oligodendroblastus immature oligodendrocyte and mature oligodendrocyte stage was observed. Results: compared with GPR56 r-mice, the formation of myelin sheath in the corpus callosum of P14 P21 and P28 GPR56-/-mice was significantly decreased. Electron microscope showed that the number of myelinated axons in the corpus callosum of P28 GPR56-r- mice decreased significantly, the g-ratio value of myelin sheath became larger and the thickness of myelin sheath became thinner. The results of fluorescence immunohistochemistry and in situ hybridization showed that there was no difference in the number of PDGF-aR cells in the corpus callosum between P7 GPR56 r- and GPR56-r- mice, but the number of PLP cells in the corpus callosum of P28 GPR56 /-mice was significantly higher than that of P28 GPR56-r- mice. The results of cell culture in vitro showed that the percentage of GPR56-/-O4 cells at pro-oligodendroblast stage was significantly higher than that of GPR56 / O4 cells, and the percentage of GPR56-/-O4 cells at immature oligodendrocyte and mature oligodendrocyte stage was significantly lower than that of GPR56 / O4 cells. Conclusion the protein of GPR56 may be involved in myelinization of white matter and maturation of OPCs.
【作者单位】：广东省人民医院广东省医学科学院急危重症医学部;
【基金】：国家自然科学基金资助项目(No.81271329)
【分类号】：R363

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