Recent advances in fish cytogenetics have enhanced the interest in chromosome analysis in both fundamental (systematics and comparative genomics among fishes and other vertebrate groups) and applied (aquaculture, conservation and response to pollutants, whole genome sequencing of model fish species) research. Although the genomic material, the chromosomes, is basically the same in the various organisms, experience has clearly shown that fish chromosomes have to be handled with specific protocols.
In laboratories around the world, traditional cytogenetic techniques and molecular cytogenetics have seen a vast improvement but the technical details and protocols are often not published in scientific journals because of constraints in format and size and therefore they are not accessible to the whole scientific community. These shortcomings are very well known to fish cytogeneticists and the idea to prepare a specific manual was first discussed during the first "Workshop of Fish Cytogenetics" held in Concarneau (France), in 1992. Resulting from the above workshop, a draft compilation of the techniques and protocols in fish cytogenetics available at that time had been produced and distributed privately, but never published. There does not exist, at present, any published handbook or manual specifically dealing with fish cytogenetics.
This book fills this void by bringing together and updating the main techniques so that the traditional methods can be standardized. Moreover, this book includes novel developments or improvements; some of the protocols provided here are practically unknown to the scientific community and even now unpublished.
Table of Contents
Teleost Fish Handling and Transport under Reduced Stress Conditions. Storage of Karyotyped Voucher Specimens and their Molecular Identification. Direct Chromosome Preparation from Freshwater Teleost Fishes. Direct Mitotic Chromosome Preparations from Chondrichthyan Tissues. Mitotic Chromosome Preparations of Freshwater Stingrays. Direct Chromosome Preparation from Regenerating Fish Fin Tissue. Direct Chromosome Preparations from Embryos and Larvae. Establishment of Sturgeon Primary Cell Lines. Teleost Fish Lymphocyte Culture. Rapid Fibroblast Culture for Teleost Fish Karyotyping. Cephalic Kidney and Spleen Cell Culture in Antarctic Teleosts. Meiotic Surface-spread and Immunodetection for Synaptonemal Complex Proteins in Teleost fish. C-banding. Sequential Chromosome Banding in Fishes. Standard Fluorescence in Situ Hybridization Procedures. Characterization of Fish Genomes by GISH and CGH. Fish Chromosome Microdissection and its Utilization in the Synthesis of Fluorescent in situ Hybridization Probes. Microdissection and Chromosome Painting. Extending DNA Fibers and Fiber-FISH. In vivo Micronucleus Test for Detecting Response of Teleost Fish to Environmental Genotoxic Exposure.
Catherine Ozouf-Costaz, Eva Pisano, Fausto Foresti, Lurdes Foresti de Almeida Toledo