International Mammalian Genome Society

17th International Mouse Genome Conference

9-12 November 2003, Braunschweig, Germany


Kusakabe M
1) ANB Tsukuba Institute, 2) Institute for Animal Reproduction

Co-Authors: Inoue J, Aotsuka S, Inoue T, Matsuba K
Institutions: ANB Tsukuba Institute

After the recent genome project was finished, many researchers' interests were moved to the functional genome analysis. Although cDNA micro array and RT-PCR methodologies are very sensitive to detect a lower level of gene expression, they never show us in situ localization of mRNA. Therefore, it is very important not only to know which cell expresses a gene, but also to know when and where they express a gene. For this purpose, in situ hybridization method can be useful tool. Although the functional molecule is not always localized within the producing cell, we can know the histological location of mRNA by in situ hybridization. Therefore, we have tried to develop the large scale in situ hybridization technique.

First, we designed the temperature controllable metal block that installed a metal bar for agitating the hybridization solution. Second, we developed the supportive computer program for the probe preparation. Then, we made a Dig-labeled cRNA probe by this method. Finally, in situ hybridization was carried out using our large scale in situ system. After all reactions have been done, the section was observed by laser microscope. Then, we made a whole image of the section by the tiling of images. Subsequently, a three-dimensional image was reconstructed by these whole images.

In conclusion, it seems that our large scale in situ hybridization system makes us possible to develop the fine histological map of gene expression.

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