Functional analysis of BDNF protein using Proteomics techinology

利用蛋白质组学技术对 BDNF 蛋白进行功能分析

• 批准号: 18500259
• 负责人: KOJIMA Masami
• 金额: $ 2.63万
• 依托单位: National Institute of Advanced Industrial Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18500259/
• 关键词: Neuron; Protein; DNA; Brain; 神経科学; 脂質; 脳・神経

ProBDNF, through activation of p75NTR, exerts functions opposite to those of mature BDNF (mBDNF). Thus, proteolytic conversion of proBDNF to mBDNF determines the direction of BDNF actions. Here we characterize two rare human SNPs (R125M and R127L) that substitute two arginines adjacent to the cleavage site. The SNPs markedly inhibited the endo-proteolytic cleavage of proBDNF. The resulting cleavage-resistant proBDNF facilitated apoptosis of cerebellar granule neurons (CGNs) and suppressed neurite outgrowth of cultured basal forebrain cholinergic neurons (BFCNs). Knock-in mice (BDNF pro/pro) with the arginine substitutions exhibited a smaller cerebellum, enhanced CGN apoptosis and decreased proliferation of CGN precursors. In basal forebrain, inhibition of proBDNF cleavage did not affect BFCNs survival but suppressed cholinergic afferents to the hippocampus. These effects were caused by an increase in proBDNF rather than the decrease in mBDNF, since crossing BDNF pro/pro with p75NTR mutant mice rescued the cellular phenotypes. These results reveal the in vivo function of proBDNF and underscore the importance of proBDNF cleavage.

ProBDNF通过激活p75NTR发挥与成熟BDNF （mBDNF）相反的功能。因此，proBDNF向mBDNF的蛋白水解转化决定了BDNF的作用方向。在这里，我们描述了两个罕见的人类snp (R125M和R127L)，它们替代了靠近切割位点的两个精氨酸。这些snp显著抑制了proBDNF的内蛋白水解裂解。所得抗裂proBDNF促进了小脑颗粒神经元（CGNs）的凋亡，抑制了培养的基底前脑胆碱能神经元（BFCNs）的神经突生长。精氨酸取代的敲入小鼠（BDNF pro/pro）表现出小脑变小、CGN凋亡增强和CGN前体增殖减少。在基底前脑中，抑制proBDNF切割不影响BFCNs的存活，但抑制了向海马的胆碱能传入。这些影响是由proBDNF的增加而不是mBDNF的减少引起的，因为将BDNF pro/pro与p75NTR突变小鼠杂交可以挽救细胞表型。这些结果揭示了proBDNF在体内的功能，并强调了proBDNF切割的重要性。

项目成果

Mutant huntintin impairs the post-Golgi trafficking of brain-derived neurotrophic factor but not its Val66Met polymorphism

突变亨廷顿蛋白会损害脑源性神经营养因子的后高尔基体运输，但不会损害其 Val66Met 多态性

• 发表时间: 2006
• 期刊: The Journal of Neuroscience 26
• 作者: Daniel;d.T. et al.
• 通讯作者: d.T. et al.

Brain-derive d neurotrophic factor (BDNF) stimulates cholesterol biosynthesis and accumulates presynaptic protems in lipid rafbs:A novel role for BDNF in synapse development

脑源性神经营养因子 (BDNF) 刺激胆固醇生物合成并在脂质 RAFB 中积累突触前蛋白质：BDNF 在突触发育中的新作用

• 发表时间: 2007
• 期刊: J Neumsci 27
• 作者: Suzuki S;.Kiyosue.K;Hazama;S;Ogura A;Kashihara M;Hara T;Koshmizu H;Kojma M
• 通讯作者: Kojma M