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腺相关病毒 2/9 介导 Cre 重组酶在小鼠初级传入神经中的表达。

Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents.

机构信息

Département de pharmacologie-physiologie, Université de Sherbrooke, Sherbrooke, Québec, Canada.

Institut de pharmacologie de Sherbrooke, Université de Sherbrooke, Sherbrooke, Québec, Canada.

出版信息

Sci Rep. 2018 May 9;8(1):7321. doi: 10.1038/s41598-018-25626-y.

Abstract

Genetically-modified animal models have significantly increased our understanding of the complex central nervous system circuits. Among these models, inducible transgenic mice whose specific gene expression can be modulated through a Cre recombinase/LoxP system are useful to study the role of specific peptides and proteins in a given population of cells. In the present study, we describe an efficient approach to selectively deliver a Cre-GFP to dorsal root ganglia (DRG) neurons. First, mice of different ages were injected in both hindpaws with a recombinant adeno-associated virus (rAAV2/9-CBA-Cre-GFP). Using this route of injection in mice at 5 days of age, we report that approximately 20% of all DRG neurons express GFP, 6 to 8 weeks after the infection. The level of infection was reduced by 50% when the virus was administered at 2 weeks of age. Additionally, the virus-mediated delivery of the Cre-GFP was also investigated via the intrathecal route. When injected intrathecally, the rAAV2/9-CBA-Cre-GFP virus infected a much higher proportion of DRG neurons than the intraplantar injection, with up to 51.6% of infected lumbar DRG neurons. Noteworthy, both routes of injection predominantly transduced DRG neurons over spinal and brain neurons.

摘要

基因修饰动物模型极大地提高了我们对复杂中枢神经系统回路的理解。在这些模型中,通过 Cre 重组酶/LoxP 系统可调节特定基因表达的诱导型转基因小鼠,可用于研究特定肽和蛋白质在特定细胞群体中的作用。在本研究中,我们描述了一种有效方法,可将 Cre-GFP 选择性递送至背根神经节 (DRG) 神经元。首先,不同年龄的小鼠在双后足中注射重组腺相关病毒 (rAAV2/9-CBA-Cre-GFP)。通过在 5 天大的小鼠中使用这种注射途径,我们报告说,在感染后 6 到 8 周,约 20%的所有 DRG 神经元表达 GFP。当病毒在 2 周龄时给药时,感染水平降低了 50%。此外,还通过鞘内途径研究了病毒介导的 Cre-GFP 传递。鞘内注射时,rAAV2/9-CBA-Cre-GFP 病毒感染的 DRG 神经元比例远高于足底内注射,感染的腰椎 DRG 神经元比例高达 51.6%。值得注意的是,两种注射途径主要转导 DRG 神经元,而不是脊髓和脑神经元。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cdc/5943452/1d410ea4b6bb/41598_2018_25626_Fig1_HTML.jpg

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