Retrograde and anterograde labeling of cerebellar afferent projection by the injection of recombinant adenoviral vectors into the mouse cerebellar cortex
Retrograde and anterograde labeling of cerebellar afferent projection by the injection of recombinant adenoviral vectors into the mouse cerebellar cortex
复制标题
通过将重组腺病毒载体注射到小鼠小脑皮质中来逆行和顺行标记小脑传入投射
DOI：
10.1007/s004290050105
复制
发表时间：
1997
期刊：
影响因子：
--
通讯作者：
H. Okado
中科院分区：
文献类型：
--
作者：
T. Terashima;A. Miwa;Y. Kanegae;I. Saito;H. Okado
摘要：腺病毒载体最近被认为是将基因传递到各种组织的高效系统。我们证明，通过腺病毒引入神经末梢的报告基因可用于逆行标记细胞体，然后在体内顺行标记受感染神经元的轴突和神经末梢。我们将携带大肠杆菌β-半乳糖苷酶基因（lacZ）的复制缺陷型重组腺病毒注射到成年小鼠的小脑皮质中。逆行标记的第一个证据是在感染后 2 天获得的，当时桥脑核和网状被盖核中的神经元微弱表达 β-半乳糖苷酶，而在感染后 3 天，所有小脑前核中的神经元（已知投射到小脑皮质）均以类似高尔基体的方式被 X-gal 强烈染色。 lacZ基因产物的顺行运输在感染后3天被识别； β-半乳糖苷酶阳性轴突源自逆行标记神经元的体细胞或树突，穿过小脑中脚或下脚，进入小脑。感染后 8 天在注射侧和感染后 14 天在对侧识别出顺行标记的苔藓末端。 β-半乳糖苷酶表达持续长达两个月，标记细胞总数随着时间的推移而减少。在注射后直至感染后 58 天的任何时间点，我们在与小脑皮质突触相连的细胞核中均未发现任何顺行或逆行突触标记的迹象。
Abstract Adenoviral vectors have recently been recognized as highly efficient systems for gene delivery into various tissues. We show that a reporter gene introduced into nerve terminals via an adenovirus can be used to label cell bodies retrogradely and then label the axons and nerve terminals of the infected neurons anterogradely in vivo. We injected a replication-defective recombinant adenovirus carrying the E. coli β-galactosidase gene (lacZ) into the cerebellar cortex of the adult mouse. The first evidence of retrograde labeling was obtained at 2 days after the infection when neurons in the pontine nuclei and the reticulotegmental nucleus of the pons weakly expressed β-galactosidase, and at 3 days post-infection when neurons in all precerebellar nuclei, known to project to the cerebellar cortex, were strongly stained with X-gal in a Golgi-like manner. Anterograde transport of lacZ gene products was recognized at 3 days post-infection; β-galactosidase-positive axons arose from somata or dendrites of retrogradely labeled neurons, passed through the middle or inferior cerebellar peduncles, and entered the cerebellum. Anterogradely labeled mossy terminals were recognized on the injection side at 8 days post-infection, and on the contralateral side at 14 days post-infection. β-Galactosidase expression persisted for up to two months, with a decrease in the total number of labeled cells over time. We could not find any signs of anterograde or retrograde transsynaptic labeling in the nuclei synaptically linked to the cerebellar cortex at any time point after injection up to 58 days post-infection.
均匀的橄榄小脑传导时间是大鼠小脑中浦肯野细胞复合体尖峰同步性的基础。
DOI：
10.1113/jphysiol.1993.sp019857
发表时间：
1993
期刊：
The Journal of physiology
影响因子：
--
作者：
Sugihara,I;Lang,EJ;Llinás,R
通讯作者：
Llinás,R