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腺病毒和腺相关病毒载体介导的大鼠脑心血管控制区域神经元基因转移。

Adenoviral and adeno-associated viral vectors-mediated neuronal gene transfer to cardiovascular control regions of the rat brain.

机构信息

School of Biotechnology, Southern Medical University, Guangzhou, China.

出版信息

Int J Med Sci. 2013;10(5):607-16. doi: 10.7150/ijms.5700. Epub 2013 Mar 20.

DOI:10.7150/ijms.5700
PMID:23569423
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3619099/
Abstract

Viral vectors have been utilized extensively to introduce genetic material into the central nervous system. In order to investigate gene functions in cardiovascular control regions of rat brain, we applied WPRE (woodchuck hepatitis virus post-transcriptional regulatory element) enhanced-adenoviral (Ad) and adeno-assoicated virus (AAV) type 2 vectors to mediate neuronal gene delivery to the paraventricular nucleus of the hypothalamus, the nucleus tractus solitarius and the rostral ventrolateral medulla, three important cardiovascular control regions known to express renin-angiotensin system (RAS) genes. Ad or AAV2 harboring an enhanced green fluorescent protein (EGFP) reporter gene or the angiotensin type 2 receptor gene were microinjected into these brain regions in adult rats. Our results demonstrated that both AAV2 and Ad vectors elicited long-term neuronal transduction in these regions. Interestingly, we found that the WPRE caused expression of GFP driven by the synapsin1 promoter in pure glial cultures or co-cultures of neurons and glia derived from rat hypothalamus and brainstem. However, in rat paraventricular nucleus WPRE did not cause expression of GFP in glia. This demonstrates the potential use of these vectors in studies of physiological functions of certain genes in the cardiovascular control regions of the brain.

摘要

病毒载体已被广泛用于将遗传物质导入中枢神经系统。为了研究大鼠脑心血管控制区的基因功能，我们应用 WPRE（土拨鼠肝炎病毒转录后调控元件）增强型腺病毒（Ad）和腺相关病毒（AAV）2 型载体将神经元基因递送至下丘脑室旁核、孤束核和延髓头端腹外侧区，这三个已知表达肾素-血管紧张素系统（RAS）基因的重要心血管控制区。携带增强型绿色荧光蛋白（EGFP）报告基因或血管紧张素Ⅱ型受体基因的 Ad 或 AAV2 被微注射到成年大鼠的这些脑区。我们的结果表明，AAV2 和 Ad 载体在这些区域均能引起长期的神经元转导。有趣的是，我们发现 WPRE 可引起源自大鼠下丘脑和脑干的神经元和神经胶质共培养物或纯神经胶质培养物中突触素 1 启动子驱动的 GFP 的表达。然而，在大鼠室旁核中，WPRE 并未引起神经胶质中 GFP 的表达。这表明这些载体可用于研究脑心血管控制区某些基因的生理功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/d55f22193a26/ijmsv10p0607g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/54768538b0bf/ijmsv10p0607g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/ad4f89b13377/ijmsv10p0607g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/f9e759fe6922/ijmsv10p0607g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/65904f5e680d/ijmsv10p0607g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/2a0ae8552bbc/ijmsv10p0607g06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/d55f22193a26/ijmsv10p0607g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/54768538b0bf/ijmsv10p0607g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/ad4f89b13377/ijmsv10p0607g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/f9e759fe6922/ijmsv10p0607g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/65904f5e680d/ijmsv10p0607g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/2a0ae8552bbc/ijmsv10p0607g06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/643f/3619099/d55f22193a26/ijmsv10p0607g09.jpg

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