Stimulus-triggered enhancement of chilling tolerance in zebrafish embryos.

BACKGROUND: Cryopreservation of zebrafish embryos is still an unsolved problem despite market demand and massive efforts to preserve genetic variation among numerous existing lines. Chilled storage of embryos might be a step towards developing successful cryopreservation, but no methods to date have worked. METHODS: In the present study, we applied a novel strategy to improve the chilling tolerance of zebrafish embryos by introducing a preconditioning hydrostatic pressure treatment to the embryos. In our experiments, 26-somites and Prim-5 stage zebrafish embryos were chilled at 0°C for 24 hours after preconditioning. Embryo survival rate, ability to reach maturation and fertilizing capacity were tested. RESULTS: Our results indicate that applied preconditioning technology made it possible for the chilled embryos to develop normally until maturity, and to produce healthy offspring as normal, thus passing on their genetic material successfully. Treated embryos had a significantly higher survival and better developmental rate, moreover the treated group had a higher ratio of normal morphology during continued development. While all controls from chilled embryos died by 30 day-post-fertilization, the treated group reached maturity (~90-120 days) and were able to reproduce, resulting in offspring in expected quantity and quality. CONCLUSIONS: Based on our results, we conclude that the preconditioning technology represents a significant improvement in zebrafish embryo chilling tolerance, thus enabling a long-time survival. Furthermore, as embryonic development is arrested during chilled storage this technology also provides a solution to synchronize or delay the development.

Mitochondrial DNA variability in Gyimesi Racka and Turcana sheep breeds.

The current knowledge and documentation on the origins and relationship between Gyimesi Racka reared in Hungary and the Romanian Turcana is rather controversial. Lack of information and scientific reliable proofs for the divergent theories found in the two countries motivated us to implement a trial using molecular methods to assess the genetic distance and diversity in the two breeds. Hair follicles were collected from Gyimesi Racka (2 phenotypes) and from Turcana (6 ecotypes). The 599 bp segment of the D-loop region of the mitochondrial DNA was sequenced. Altogether, 42 haplotypes were identified, while 23 were found in both populations. Populations were highly diverse according to the haplotype and nucleotide diversity indices. AMOVA analysis showed that most of the variation was observed within populations (98%), indicating a weak genetic structure between the two breeds. Animals were grouped into seven groups based on their phenotype; however genetic distances among them were also low. Tajima's D, Fu's Fs, goodness-of-fit statistics, mismatch distribution and network analysis suggested recent demographic expansion. Current comprehensive mtDNA study indicates that there is very low level of genetic differentiation between the Gyimesi Racka and Turcana populations therefore they are de facto one trans-boundary breed.