Antisense in vivo knockdown of synaptotagmin I and synapsin I by HVJ-liposome mediated gene transfer modulates ischemic injury of hippocampus in opposing ways
Antisense in vivo knockdown of synaptotagmin I and synapsin I by HVJ-liposome mediated gene transfer modulates ischemic injury of hippocampus in opposing ways
复制标题
DOI：
10.1016/s0168-0102(02)00233-x
复制
发表时间：
2003-03-01
影响因子：
2.9
通讯作者：
Saji, M
中科院分区：
文献类型：
--
作者：
Iwakuma, M;Anzai, T;Saji, M
缺血性脑损伤的发病机制是兴奋性毒性，而兴奋性毒性是由谷氨酸受体过度激活和钙超载介导的。为探讨神经末梢释放递质在促进缺血性神经元变性中的作用，我们采用HVJ-脂质体基因转移技术，将突触结合蛋白I（synaptotagmin I）或突触素I（synapsin I）的反义寡核苷酸（ODNs）导入大鼠脑内。在短暂性前脑缺血20 min前4 d，将反义ODNs注射到大鼠侧脑室，单次反义处理可使突触终末递质释放相关蛋白水平持续下调。突触结合蛋白I的反义基因敲除几乎完全阻止了海马CA 1区神经元的缺血性损伤，而突触结合蛋白I的反义基因敲除则明显促进了海马CA 1区锥体神经元的缺血性损伤，并将损伤扩展到相对耐受的CA 2/CA 3区。通过体内敲低突触结合蛋白I或突触蛋白I对缺血性海马损伤的调制表明，从末端释放的递质在缺血性脑损伤的演变中起重要作用，因此通过使用反义ODNs-HVJ-脂质体复合物的递质释放策略对于神经保护疗法是可靠的。(C)2002 Elsevier Science爱尔兰有限公司和日本神经科学学会。All rights reserved.
Neurotransmitter release during and after ischemic event is thought to be involved in excitotoxicity as a pathogenesis for the ischemic brain damage, which is mediated by excessive activation of glutamate receptors and attendant calcium overload. To ascertain the role of transmitter release from nerve terminals in promoting the,ischemic neurodegeneration, we delivered antisense oligodeoxynucleotides (ODNs) to synaptotagmin I or synapsin I into the rat brain by using HVJ-liposome gene transfer technique. The antisense ODNs were injected into the lateralventricle in rats 4 days prior to transient forebrain ischemia of 20 min. With a single antisense treatment, long-lasting downregulation of the transmitter release relating protein levels at overall synaptic terminals was achieved. The antisense in vivo knockdown of synaptotagmin I prevented almost completely the ischemic damage of hippocampal CA1 neurons, while the in vivo knockdown of synapsin I markedly promoted the ischemic damage of CA1 pyramidal neurons and extended the injury to relatively resistant CA2/CA3 region. The modulation of ischemic hippocampal damage by the in vivo knockdown of synaptotagmin I or synapsin I suggests that transmitter release from terminals plays an important role in the evolution of ischemic brain damage and therefore the transmitter release strategy by the use of antisense ODNs-HVJ-liposome complex is reliable for neuroprotective therapies. (C) 2002 Elsevier Science Ireland Ltd and the Japan Neuroscience Society. All rights reserved.