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Nucleic Acids Res February 1, 2008; 36 (2): 387-92.

Intracellular expression profiles measured by real-time PCR tomography in the Xenopus laevis oocyte.

Sindelka R , Jonák J , Hands R , Bustin SA , Kubista M .

Real-time PCR tomography is a novel, quantitative method for measuring localized RNA expression profiles within single cells. We demonstrate its usefulness by dissecting an oocyte from Xenopus laevis into slices along its animal-vegetal axis, extracting its RNA and measuring the levels of 18 selected mRNAs by real-time RT-PCR. This identified two classes of mRNA, one preferentially located towards the animal, the other towards the vegetal pole. mRNAs within each group show comparable intracellular gradients, suggesting they are produced by similar mechanisms. The polarization is substantial, though not extreme, with around 5% of vegetal gene mRNA molecules detected at the animal pole, and around 50% of the molecules in the far most vegetal section. Most animal pole mRNAs were found in the second section from the animal pole and in the central section, which is where the nucleus is located. mRNA expression profiles did not change following in vitro fertilization and we conclude that the cortical rotation that follows fertilization has no detectable effect on intracellular mRNA gradients.

PubMed ID: 18039714
PMC ID: PMC2241880
Article link: Nucleic Acids Res

Species referenced: Xenopus laevis
Genes referenced: ctnnb1 dazl dvl2 foxh1.2 gdf1 otx1 pou5f3.3 tcf3 vegt wnt11

Article Images: [+] show captions
References [+] :
Bengtsson, Gene expression profiling in single cells from the pancreatic islets of Langerhans reveals lognormal distribution of mRNA levels. 2005, Pubmed