在Cre/loxP基因重组小鼠脑内植入“光控开关”建立偏头痛模型

项目名称： 在Cre/loxP基因重组小鼠脑内植入“光控开关”建立偏头痛模型

项目编号： No.81471147

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 于生元

作者单位： 中国人民解放军总医院

项目金额： 80万元

中文摘要： 缺少可靠的动物模型是偏头痛研究的难点。虽然家族性偏瘫型偏头痛（familial hemiplegic migraine, FHM）致病基因敲入（knock-in，KI）小鼠是带来希望的工具，但FHM基因编码的离子通道既在全身广泛分布又有高度组织特异性，多数致病基因的纯合KI小鼠死于胚胎期，而杂合动物的偏头痛表现不明显，造模方法仍需改良。本项目以偏头痛的兴奋/抑制稳态失衡学说为依据，一方面将光敏蛋白引入几种标记不同类型神经元的Cre重组酶小鼠中，通过直接光控特定脑区某类神经元活性，构建偏头痛易感性增强小鼠模型；另一方面利用Cre/loxP基因重组技术培育FHM3型基因条件性敲入小鼠，对其植入光敏蛋白，光控调节其神经元活性来缓解基因突变所致脑内兴奋/抑制稳态失衡，以构建慢性自发性偏头痛小鼠模型。从而为更加深入研究偏头痛发病机制，为新药研发提供更为有效、可靠、更接近偏头痛临床表现的动物模型。

中文关键词： 偏头痛；光遗传学；神经电生理；离子通道；基因重组

英文摘要： The lack of reliable animal models has been the very obstacle of studying the mechanisms of migraine and the effect of acute and phylactic migraine drugs. Although the knock-in mice with the familial hemiplegic migraine genes became the hoping tools for migraine studying, the corresponding ion channels, which distribute all over the system, express specifically in different organs or tissue with different level. Most of the homozygous knock-in mice with FHM mutations die at birth or on postnatal day, while the heterozygous mice with FHM mutations do not exhibit an overt phenotype. It is important and urgent to develop more reliable migraine animal model. In our study, based on the theory that migraine is a disorder of brain excitatory-inhibitory balance, we plan to develop two types of mouse model, involving the combination of optogenetics and Cre/loxP genetic recombination. On one side, we are going to induce bi-stable opsin ChR2-SSFO into two types of neurons in different brain regions of two lines of Cre- recombinase mice, to activate or inactivate neural activity by manipulating the opsins with light in millisecond precision, in order to develop an increasing susceptibility model of migraine. On the other side, a conditional genetic knock-in mouse model with FHM3 mutation (Q1489K) will be developed, involving the loxP recombination technique. A ChR2 （H143R）opsin will be transplanted at the same time, hoping that the opsin will stabilize the excitatory/inhibitory imbalance to prolong the survival life of the mutant mice, which will be a model of chronic spontaneous migraine attacks. We hope that both of the new animal models can be very efficient and powerful for the studying of migraine in the future.

英文关键词： migraine;optogenetics;eletrophysiology;ion channel;gene recombination