CAJ | 학술논문

目的：探讨G蛋白偶联受体56（G-protein coupled receptor 56，GPR56）对小鼠脑胼胝体白质内轴突发育和髓鞘化的影响。方法筛选出 Gpr56基因杂合型（Gpr56＋/-）和敲除型（Gpr56-/-）小鼠64只，随机数字法分为2组：Gpr56＋/-和Gpr56-/-组，每组32只。每组根据小鼠出生后时间分为出生后7 d、14 d、21 d、28 d （P7 d、P14 d、P21 d、P28 d）四个亚组。应用免疫组化染色和Western Blot方法监测神经纤维丝蛋白（NF-200）、髓鞘蛋白脂质蛋白（PLP）在出生后P7 d、P14 d、P21 d、P28 d 的 Gpr56＋/-和 Gpr56-/-小鼠脑胼胝体白质中的表达；用 P1d Gpr56＋/-和Gpr56-/-小鼠皮质做体外神经元培养，观察两组神经元轴突的长短；用电镜观察P28d Gpr56＋/-和Gpr56-/-小鼠胼胝体白质内轴突髓鞘化，比较轴突直径的大小。结果与Gpr56＋/-小鼠比较，NF-200和 PLP蛋白在P14 d、P21 d、P28 d 的Gpr56-/-小鼠脑胼胝体白质中的表达明显下降。Gpr56-/-神经元轴突明显短于Gpr56＋/-神经元。电镜见P28d Gpr56-/-小鼠脑胼胝体白质内髓鞘化轴突的数量明显减少，轴突直径明显变小。结论 GPR56蛋白分子可能参与了脑白质轴突轴突发育和髓鞘化。
목적：탐토G단백우련수체56（G-protein coupled receptor 56，GPR56）대소서뇌변지체백질내축돌발육화수초화적영향。방법사선출 Gpr56기인잡합형（Gpr56＋/-）화고제형（Gpr56-/-）소서64지，수궤수자법분위2조：Gpr56＋/-화Gpr56-/-조，매조32지。매조근거소서출생후시간분위출생후7 d、14 d、21 d、28 d （P7 d、P14 d、P21 d、P28 d）사개아조。응용면역조화염색화Western Blot방법감측신경섬유사단백（NF-200）、수초단백지질단백（PLP）재출생후P7 d、P14 d、P21 d、P28 d 적 Gpr56＋/-화 Gpr56-/-소서뇌변지체백질중적표체；용 P1d Gpr56＋/-화Gpr56-/-소서피질주체외신경원배양，관찰량조신경원축돌적장단；용전경관찰P28d Gpr56＋/-화Gpr56-/-소서변지체백질내축돌수초화，비교축돌직경적대소。결과여Gpr56＋/-소서비교，NF-200화 PLP단백재P14 d、P21 d、P28 d 적Gpr56-/-소서뇌변지체백질중적표체명현하강。Gpr56-/-신경원축돌명현단우Gpr56＋/-신경원。전경견P28d Gpr56-/-소서뇌변지체백질내수초화축돌적수량명현감소，축돌직경명현변소。결론 GPR56단백분자가능삼여료뇌백질축돌축돌발육화수초화。
Objective To determine the likelihood of G-protein coupled receptor 56 (GPR56 ) induces axonal development and myelination in the corpus callosum of mouse brain.Methods A total of 64 Gpr56 +/-and Gpr56 -/-mice were selected and randomly divided into two groups:Gpr56 +/-group (n=32)and Gpr56 -/-group (n=32).According to number of days after birth,each group was further divided into 4 subgroups including P7d,P14d,P21d and P28d subgroups.Levels of neurofilament-200 (NF -200)and proteolipid protein (PLP ) of myelin basic protein in corpus callosum were measured with immunohistochemistry staining and Western blot in P7d、P14d、P21d、P28d Gpr56 +/- and Gpr56 -/-mice.Gpr56 +/-and Gpr56 -/-neurons were cultured using P1 d Gpr56 +/-and Gpr56 -/-mouse brain.The lengths of Gpr56 +/- and Gpr56 -/-neuronal axon were measured and compared with Image J software. Axonal myelination in the corpus callosum of mouse brain in each group was observed under electronic microscopy and the axonal diameters between subgroups were compared.Results The levels of NF-200 and PLP in the corpus callosum in P7d、P14d、P21d、P28d Gpr56 -/-mice decreased significantly compared with Gpr56 +/- mice.The length of Gpr56 -/-neuronal axon was shortened compared with Gpr56 +/-neuronal axon.The number of myelinated axons was obviously reduced in the corpus callosum in P28d Gpr56 -/-mice.The diameter of axon in the corpus callosum of P28d Gpr56 +/-mouse is longer than that of P28d Gpr56 -/-mouse. Conclusions GPR56 may be involved in axonal development and myelination in the corpus callosum of mouse brain.