Fax +41 61 306 12 34 E-Mail karger@karger.ch www.karger.com Review Article Cytogenet Genome Res 2009;126:34–48 DOI: 10.1159/000245905 Fluorescence in situ Hybridization Applied to Domestic Animal Cytogenetics J. Rubes a A. Pinton b A. Bonnet-Garnier b V. Fillon b P. Musilova a K. Michalova c S. Kubickova a A. Ducos b M. Yerle b a Veterinary Research Institute, Brno, Czech Republic; b UMR 444 Génétique Cellulaire, INRA-ENVT, Toulouse, France; c General Faculty Hospital and 1st Faculty of Medicine Charles University, Prague, Czech Republic Molecular Cytogenetics Application in Diagnostics and Research Methods of in situ hybridization in cytogenetics are based on the ability of single-stranded nucleic acids to bind with complementary parts of the target DNA, which is fixed on a microscope slide. Target DNA must be single stranded as well and DNA probe is labeled either ra- dioactively or with some fluorochromes. Preferentially, non-radioactively labeled DNA has been used as probe for in situ hybridization, and localization of probes is per- formed by indirect or direct immunofluorescence. Fluo- rescent signals are detected using a fluorescence micro- scope, equipped with a high-performance CCD (charge coupled device) camera and software specified for chro- mosome and signal analysis. Several fluorochromes can be used for simultaneous evaluation of hybridization sig- nals in one experiment. Different kinds of DNA probes can be used in hybridization experiments such as centro- mere and locus-specific probes, painting probes for whole chromosomes or their parts, or telomere and subtelomere probes. The type of DNA probe most suitable for in situ hybridization is determined by the specificity of the clin- ical diagnosis or conventional cytogenetics. Fluorescence in situ hybridization (FISH) has been de- veloped and extensively used to: Key Words Cattle  Cytogenetics  FISH  Horse  Pig  Poultry Abstract The aim of this article is not to present an exhaustive review of molecular cytogenetics applications in domestic animal species, but more to illustrate the considerable contribution of these approaches in diagnostics and research in econom- ically important species. A short presentation of the main applications of molecular cytogenetics in humans points out the domains in which it has become an essential tool and underlines the specificities attached to this species in com- parison to farm animals. This article is devoted to outlining the current resources available in domestic species and to some examples of fluorescence in situ hybridization applica- tions in the cattle, pig, horse and avian species. From a clini- cal point of view, these examples illustrate the advantages of FISH for the study of chromosomal abnormalities (identi- fication, characterization and estimation of their effects). Other applications of molecular cytogenetics are also illus- trated, particularly ZOO-FISH, an approach which allows the determination of chromosome homologies between spe- cies. Finally, a specific emphasis was placed on the useful- ness of molecular cytogenetics for the analysis of species such as poultry, which harbour a complex karyotype. Copyright © 2009 S. Karger AG, Basel Accepted after revision: July 15, 2009 by M. Schmid Jiri Rubes Veterinary Research Institute Hudcova 70, 621 32 Brno (Czech Republic) Tel. +420 5 3333 1401, Fax +420 5 4121 1229 E-Mail rubes @ vri.cz © 2009 S. Karger AG, Basel 1424–8581/09/1262–0034$26.00/0 Accessible online at: www.karger.com/cgr