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目的:探讨脂筏对于GDNF与其膜受体相互作用的影响。方法:MN9D细胞和SD大鼠随机分成三组:GDNF组、PBS组、空白对照组。两种方式处理三组样品:破坏和不破坏细胞膜上的脂筏。通过免疫共沉淀和免疫印迹检测破坏脂筏前后GDNF及其配体结合受体GFRα1与四种膜受体(RET,NCAM140,integrinβ1和N-cadherin)之间的结合情况。结果:细胞膜上的脂筏未被破坏前,四种膜受体都能够与GDNF结合发生免疫共沉淀。脂筏被破坏后,GDNF只能与RET和N-cadherin结合发生免疫共沉淀,并且这种结合在GDNF组中最多;而GFRα1只能和RET结合发生免疫共沉淀。结论:膜受体介导的GDNF生物学作用均可发生在脂筏这一平台上,脂筏在GDNF与其受体相互作用中扮演着重要角色。
Objective: To investigate the effect of lipid raft on the interaction between GDNF and its membrane receptors. Methods: MN9D cells and SD rats were randomly divided into three groups: GDNF group, PBS group and blank control group. Three sets of samples were processed in two ways: destroy and not destroy lipid rafts on the cell membrane. Co-immunoprecipitation and immunoblotting were used to detect the binding of GDNF and its ligand-binding receptor GFRα1 to four membrane receptors (RET, NCAM140, integrinβ1 and N-cadherin) before and after lipid raft disruption. RESULTS: All four membrane receptors were co-immunoprecipitated with GDNF before the lipid rafts on the cell membrane were destroyed. After the lipid raft was destroyed, GDNF could only co-immunoprecipitate with RET and N-cadherin, and this binding was the most in GDNF group. However, GFRα1 could only co-immunoprecipitate with RET. CONCLUSION: Membrane receptor-mediated biological effects of GDNF can occur on the lipid raft platform. Lipid rafts play an important role in the interaction between GDNF and its receptor.