INSERM E9935 Hôpital Robert Debré 48 Boulevard Sérurier 75019 Paris, France
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presenter: Michel Simonneau
Michel Simonneau, Fabien Guimiot, Christophe Mas, Hassan Al Halabiah, Jean-Marie
Moalic, Béatrice Levacher, Francine Bourgeois
Neurogénétique, INSERM E9935, Hôpital Robert Debré, 48 Boulevard Sérurier 75019 Paris, France
Although the number and level of expression of the entire set of genes expressed
in a given eukaryotic cell type can be assessed by high-thoughput differential
gene-expression technologies (see for instance, Velculescu et al., 1999), the
identity of a large number of the transcripts remains unknown.
Our group is working on the identification of novel genes involved in proliferation
and migration of stem cells in the embryonic future cortex (dorsal telencephalon),
which occur at early stages of embryonic development (embryonic days 10, in
the mouse). To identify such genes, we used differential display RT-PCR, which
can detect rare transcripts and generate fingerprints of mRNA populations.
We report three set of experiments:
(i) analysis of genes involved in the proliferation of stem cells by comparing proliferating cells to embryonic postmitotic cells from embryonic telencephalon;
(ii) analysis of genes involved in the migration of neuroblasts by comparing transcripts from transgenic embryos having mutations in genes involved in migration of neuroblasts with those from normal embryos;
(iii) analysis of genes regulated by a 570 kb YAC transgene from human chromosome 21 (125F7; a line generated in E.M. Rubins laboratory) which contains four Down syndrome critical genes (TTC3, DYRK1A, DSCR5, DSCR3).
More than half the differential transcripts identified in experiments (i) and (ii) have unknown functions. Some of them have no database match. We describe the characterisation of some of these transcripts.
As differential display RT-PCR allows multiple comparison, a developmental profile can be obtained for low-abundance transcripts which is not attainable by northern blotting. Furthermore, the sensitivity of the RT-PCR technique allows is discussed for the detection of small variations in low-abundance transcripts at the difference of microarray technologies.
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