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使用LacZ报告基因对小鼠Rhobtb3进行表达分析及敲除品系的初步表征。

Expression analysis of mouse Rhobtb3 using a LacZ reporter and preliminary characterization of a knockout strain.

作者信息

Lutz Julia, Grimm-Günter Eva-Maria S, Joshi Pooja, Rivero Francisco

机构信息

Centre for Cardiovascular and Metabolic Research, Hull York Medical School, University of Hull, Cottingham Road, HU6 7RX, Hull, UK.

出版信息

Histochem Cell Biol. 2014 Nov;142(5):511-28. doi: 10.1007/s00418-014-1235-9. Epub 2014 Jun 13.

DOI:10.1007/s00418-014-1235-9
PMID:24923387
Abstract

RhoBTB3 is an atypical member of the Rho family of small GTPases. It localizes at the Golgi apparatus and endosomes and is involved in vesicle trafficking and in targeting proteins for degradation in the proteasome. Previous studies using Northern blot analysis showed that Rhobtb3 is ubiquitously expressed in adult mice, but expression is particularly high in brain, heart and uterus. The gene is also expressed between embryonic days 11.5 and 17.5. To investigate the specific cell types that express this gene across tissues, both in the embryo and in the adult organism, we have made use of a gene trap mouse strain that expresses the LacZ gene under the transcriptional control of the endogenous Rhobtb3 promoter. Histochemical detection of β-galactosidase expression revealed a profile characterized by nearly ubiquitous expression of Rhobtb3 in the embryo, but with particularly high levels in bone, cartilage, all types of muscle, testis and restricted areas of the nervous system. In the adult, expression persists at much lower levels in cardiac muscle, the tunica media of blood vessels and cartilage and at high levels in the seminiferous tubules. A general preliminary characterization of this gene trap mouse strain revealed reduced viability, a postnatal growth defect and reduced testis size. Our results should pave the way for future studies aimed at investigating the roles of RhoBTB3 in tissue development and in cardiac, vascular and testicular function.

摘要

RhoBTB3是小GTP酶Rho家族的一个非典型成员。它定位于高尔基体和内体,参与囊泡运输以及将蛋白质靶向蛋白酶体进行降解。先前使用Northern印迹分析的研究表明,Rhobtb3在成年小鼠中普遍表达,但在脑、心脏和子宫中表达特别高。该基因在胚胎期11.5至17.5天之间也有表达。为了研究在胚胎和成年生物体的各种组织中表达该基因的特定细胞类型,我们利用了一种基因捕获小鼠品系,该品系在内源性Rhobtb3启动子的转录控制下表达LacZ基因。β-半乳糖苷酶表达的组织化学检测显示,Rhobtb3在胚胎中几乎普遍表达,但在骨骼、软骨、所有类型的肌肉、睾丸和神经系统的特定区域中表达水平特别高。在成年个体中,心肌、血管中膜和软骨中的表达水平持续较低,而在生精小管中表达水平较高。对这种基因捕获小鼠品系的一般初步表征显示其活力降低、出生后生长缺陷和睾丸尺寸减小。我们的结果应为未来旨在研究RhoBTB3在组织发育以及心脏、血管和睾丸功能中的作用的研究铺平道路。

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Rho family GTPases.Rho 家族 GTP 酶。
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