Cell-type specific expression of oxytocin and vasopressin genes: an experimental odyssey.
Cell-type specific expression of oxytocin and vasopressin genes: an experimental odyssey.
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DOI：
10.1111/j.1365-2826.2011.02236.x
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发表时间：
2012-04
影响因子：
3.2
通讯作者：
Gainer H
中科院分区：
文献类型：
--
作者：
Gainer H
视上核（SON）是研究细胞特异性基因表达的一个特别好的模型，因为它含有两种不同的神经元表型，催产素（OXT）和加压素（AVP）合成大细胞神经元（MCNs）。发现MCN的数量大致相等，并且在SON中约97%的MCN群体中选择性表达OXT或AVP基因。一个尚未解决的问题是，以确定什么机制是负责高度选择性调节的细胞类型特异性表达的OXT和AVP基因的MCNs。以前试图解决这个问题使用各种生物信息学和分子方法，其中包括使用异源细胞系研究推定的顺式元件中的OXT和AVP基因，和使用的OXT和/或AVP转基因在转基因啮齿动物。来自所有上述研究的数据鉴定了OXT外显子I上游<0.6kbp和AVP外显子I上游约3 kb的区域分别足以产生OXT和AVP基因的细胞特异性表达，但未能鉴定负责MCN特异性基因表达的特异性顺式结构域。一种替代的实验方法来进行启动子缺失分析在体内，即使用立体定位病毒载体基因转移到SON中，以进一步解剖的顺式元件中的OXT和AVP基因，将在这里描述。该体内方法使用表达OXT启动子缺失构建体的腺相关病毒（AAV）载体，并利用增强型绿色荧光蛋白（EGFP）作为报告基因。将AAV构建体立体定位注射到SON上方的大鼠脑中，注射后2周处死大鼠并测定EGFP表达。使用这种方法，已经可以鉴定OXT和AVP基因启动子中转录起始位点（TSS）上游的特定区域，其负责赋予SON中OXT和AVP基因表达的细胞类型特异性。
The supraoptic nucleus (SON) is a particularly good model for the study of cell-specific gene expression since it contains two distinct neuronal phenotypes, the oxytocin (OXT) and vasopressin (AVP) synthesizing magnocellular neurons (MCNs). The MCNs are found in approximately equal numbers and selectively express either the OXT or the AVP gene in about 97% of the MCN population in the SON. An unresolved issue has been to determine what mechanisms are responsible for the highly selective regulation of the cell-type specific expression of OXT and AVP genes in the MCNs. Previous attempts to address this question used various bioinformatic and molecular approaches, which included using heterologous cell lines to study the putative cis-elements in the OXT and AVP genes, and the use of OXT and/or AVP transgenes in transgenic rodents. The data from all of the above studies identified a region <0.6kbp upstream of OXT exon I and about 3kb upstream of AVP exon I as being sufficient to produce cell-specific expression of the OXT and AVP genes, respectively, but failed to identify the specific cis-domains responsible for the MCN-specific gene expression. An alternative experimental approach to perform promoter deletion analysis in vivo, that is to use stereotaxic viral vector gene transfer into the SON in order to further dissect the cis-elements in the OXT and AVP genes, will be described here. This in-vivo method uses Adeno-Associated Viral (AAV) vectors expressing OXT-promoter deletion constructs and utilizes the enhanced green fluorescent protein (EGFP) as the reporter. The AAV constructs are stereotaxically injected into the rat brain above the SON and 2 weeks post injection the rats are sacrificed and assayed for EGFP expression. Using this method it has been possible to identify specific regions upstream of the transcription start site (TSS) in the OXT and AVP gene promoters which are responsible for conferring the cell-type specificity of the OXT and AVP gene expression in the SON.
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影响因子：
2.4
作者：
de Backer, Marijke W. A.;Fitzsimons, Carlos P.;Adan, Roger A. H.
通讯作者：
Adan, Roger A. H.
影响因子：
4.8
作者：
ANG, HL;UNGEFROREN, H;MURPHY, D
通讯作者：
MURPHY, D
影响因子：
12.4
作者：
Bienemann, AS;Martin-Rendon, E;Uney, JB
通讯作者：
Uney, JB
影响因子：
4
作者：
Garza, Jacob C.;Kim, Chung Sub;Lu, Xin-Yun
通讯作者：
Lu, Xin-Yun
影响因子：
7.8
作者：
GAINER, H;SARNE, Y;BROWNSTEIN, MJ
通讯作者：
BROWNSTEIN, MJ