Lentivirus-mediated transgene delivery to the hippocampus reveals sub-field specific differences in expression.
Lentivirus-mediated transgene delivery to the hippocampus reveals sub-field specific differences in expression.
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DOI：
10.1186/1471-2202-10-2
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发表时间：
2009-01-13
期刊：
影响因子：
2.4
通讯作者：
Fitzsimons CP
中科院分区：
文献类型：
--
作者：
van Hooijdonk LW;Ichwan M;Dijkmans TF;Schouten TG;de Backer MW;Adan RA;Verbeek FJ;Vreugdenhil E;Fitzsimons CP
成年海马齿状回颗粒细胞层是由不同年龄、不同形态和不同电生理特性的神经元组成的异质结构。逆转录病毒载体已被广泛用于转导颗粒细胞层的细胞，并在完整的脑环境中研究其固有特性。此外，基于慢病毒的载体也被用来将转基因输送到复制和非复制细胞，如中枢神经系统的有丝分裂后神经元。然而，很少有研究致力于解决这些广泛使用的载体在体内对海马细胞的适用性。因此，本研究的目的是广泛地表征在海马齿状回通过VSVg伪型慢病毒载体在体内有效转导的细胞类型。在本研究中，我们使用水泡性口炎病毒G糖蛋白假型慢病毒载体在小鼠海马区表达了来自三个不同启动子的EGFP。与慢病毒转导CA1区锥体细胞相比，我们确定了齿状回颗粒细胞层中转基因表达的亚区特异性差异。此外，我们对这些慢病毒载体转导的细胞类型进行了鉴定，结果表明，它们主要针对存在于颗粒下层和颗粒细胞层更多未成熟层的神经元前体细胞和未成熟神经元。我们的观察表明，不同类型的海马细胞对慢病毒转导的通透性存在内在差异。特别是，我们首次发现，颗粒细胞层的成熟神经元不表达慢病毒传递的转基因，尽管在其他类型的海马细胞中成功表达。因此，在海马颗粒细胞中，只有成体产生的神经元是慢病毒介导的转基因转移的靶点。这些特性使慢病毒载体成为体内在小鼠海马神经祖细胞、未成熟神经元和成体生成的神经元中过度表达或敲除基因的极佳系统。
In the adult hippocampus, the granule cell layer of the dentate gyrus is a heterogeneous structure formed by neurons of different ages, morphologies and electrophysiological properties. Retroviral vectors have been extensively used to transduce cells of the granule cell layer and study their inherent properties in an intact brain environment. In addition, lentivirus-based vectors have been used to deliver transgenes to replicative and non-replicative cells as well, such as post mitotic neurons of the CNS. However, only few studies have been dedicated to address the applicability of these widespread used vectors to hippocampal cells in vivo. Therefore, the aim of this study was to extensively characterize the cell types that are effectively transduced in vivo by VSVg-pseudotyped lentivirus-based vectors in the hippocampus dentate gyrus. In the present study we used Vesicular Stomatitis Virus G glycoprotein-pseudotyped lentivirual vectors to express EGFP from three different promoters in the mouse hippocampus. In contrast to lentiviral transduction of pyramidal cells in CA1, we identified sub-region specific differences in transgene expression in the granule cell layer of the dentate gyrus. Furthermore, we characterized the cell types transduced by these lentiviral vectors, showing that they target primarily neuronal progenitor cells and immature neurons present in the sub-granular zone and more immature layers of the granule cell layer. Our observations suggest the existence of intrinsic differences in the permissiveness to lentiviral transduction among various hippocampal cell types. In particular, we show for the first time that mature neurons of the granule cell layer do not express lentivirus-delivered transgenes, despite successful expression in other hippocampal cell types. Therefore, amongst hippocampal granule cells, only adult-generated neurons are target for lentivirus-mediated transgene delivery. These properties make lentiviral vectors excellent systems for overexpression or knockdown of genes in neuronal progenitor cells, immature neurons and adult-generated neurons of the mouse hippocampus in vivo.
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影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
2.5
作者：
ALTMAN, J;BAYER, SA
通讯作者：
BAYER, SA
影响因子：
4.2
作者：
Jakobsson, J;Ericson, C;Lundberg, C
通讯作者：
Lundberg, C
DOI：
10.1073/pnas.0601992103
发表时间：
2006-05-23
影响因子：
11.1
作者：
Encinas, Juan M.;Vaahtokari, Anne;Enikolopov, Grigori
通讯作者：
Enikolopov, Grigori
影响因子：
5.1
作者：
Bonci, D;Cittadini, A;Condorelli, G
通讯作者：
Condorelli, G