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作 者:陈静[1,2] 孙羽[2] 孟艳[3] 高殿帅[2]
机构地区:[1]徐州医学院形态学实验教学中心 [2]徐州医学院神经生物学与解剖学教研室,徐州221000 [3]首都医科大学北京宣武医院,北京100069
出 处:《神经解剖学杂志》2014年第1期70-74,共5页Chinese Journal of Neuroanatomy
基 金:国家自然科学基金(31040035/C090202);江苏高校优势学科建设工程资助项目(PAPD)
摘 要: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 effects of lipid rafts in the interaction between GDNF and its membrane recep- tors. Methods: MN9D cells and SD rats were randomly divided into three groups: GDNF group, PBS group, Control group. Samples of three groups were treated with two ways : to disintegrate the lipid rafts on the cell membrane or not. Through co-immunoprecipitation and western blotting to detect the binding of GDNF or its ligand-binding receptor GFRα1 to four receptors (RET, NCAM140, integrinβ1, N-cadherin) before and after the lipid rafts were disintegrated. Results: Before the lipid rafts on the cell membrane were disintegrated, GDNF could bind to four receptors resulting in co-immuno- precipitation. After the lipid rafts were disintegrated, GDNF only could bind to RET and N-cadherin result in co-immuno- precipitation, which were highest of GDNF group. In addition, GFRα1 only could bind to RET resulting in co-immuno- precipitation. Conclusion: Our study strongly suggests the receptor-mediated biological effects of GDNF can take place on the platform of lipid rafts which play a substantial role in the interaction between GDNF and its receptors.
关 键 词:脂筏 GDNF 膜蛋白 RET NCAM140 INTEGRINΒ1 N-CADHERIN
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