Spatial expression profiles in the Xenopus laevis oocytes measured with qPCR tomography

被引：20

作者：

Sindelka, Radek ^{[2
]}

Sidova, Monika ^{[1
,3
]}

Svec, David ^{[1
]}

Kubista, Mikael ^{[1
,4
]}

机构：

[1] Acad Sci Czech Republic, Inst Biotechnol, Gene Express Lab, Prague, Czech Republic

[2] Whitehead Inst, Cambridge, MA USA

[3] Charles Univ Prague, Fac Sci, Dept Cell Biol, CR-11636 Prague 1, Czech Republic

[4] TATAA Bioctr AB, S-41103 Gothenburg, Sweden

来源：

METHODS | 2010年 / 51卷 / 01期

关键词：

Xenopus; Oocyte; qPCR tomography; Expression profiling; RNA LOCALIZATION; CELL FATE; ROTATION; EMBRYOS; TIME; AXIS; EGG;

D O I：

10.1016/j.ymeth.2009.12.011

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

qPCR tomography was developed to study mRNA localization in complex biological samples that are embedded and cryo-sectioned. After total RNA extraction and reverse transcription, the spatial profiles of mRNAs and other functional RNAs were determined by qPCR. The Xenopus laevis oocyte was selected as model, because of its large size (more than 1 mm) and large amount of total RNA (similar to 5 mu g). Fifteen sections along the animal-vegetal axis were cut and prepared for quantification of 31 RNA targets using the high-throughput real-time RT-PCR (qPCR) BioMark (TM) platform. mRNAs were found to have two localization patterns, animal/central or vegetal. Because of the high resolution in sectioning, it was possible to distinguish two subgroups of the vegetal gene patterns: germ plasm determinant pattern and profile of other vegetal genes. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：87 / 91

页数：5

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