Comparative karyotype using bidirectional chromosome painting: how and why?
Rat is widely used in biomedical and pharmaceutical research but its genome has been significantly less studied than that of the mouse. This represents a major limitation for studying cytogenetic and molecular mechanisms in the rat model. As Muridae species underwent an intense chromosome evolution it is not possible to directly transpose knowledge of the mouse genome to that of the rat. For establishing a comparative karyotype between rat and mouse, painting probes of both species were prepared by PARM-PCR (Priming Authorizing Random Mismatches PCR) from a low copy number of sorted chromosomes, the mouse and rat specific painting probes being then hybridized on rat and mouse metaphases, respectively. The availability of rodent species chromosome painting probes as well as the information obtained by the comparative karyotype and comparative gene mapping data are of great interest to improve knowledge on species evolution but also to better understand carcinogenesis process, as illustrated by our data concerning the cytogenetic characterization of radon-induced rat lung tumors. Detailed methods for obtaining painting probes by PARM-PCR from sorted mouse and rat chromosomes and for their hybridization in homologous or heterologous conditions are described. Usefulness of chromosome painting is illustrated by the characterization of chromosomal abnormalities in a radon-induced rat lung tumor. Advantages and limitations of this technique as compared to classical cytogenetics, FISH and CGH are discussed.
Guilly
M N
MN
CEA, DSV, DRR, Fontenay-aux-Roses, France. mnguilly@ellebore.saclay.cea.fr
Dano
L
L
de Chamisso
P
P
Fouchet
P
P
Dutrillaux
B
B
Chevillard
S
S
D000818:Animals; D020223:Chromosome Painting / Q000379:methods*; D007621:Karyotyping / Q000379:methods*; D008175:Lung Neoplasms / Q000235:genetics; D051379:Mice / Q000235:genetics*; D051381:Rats / Q000235:genetics*