Acronym CRYOSPERM
Category
Marine Biotechnology
Title Development of tools to analyse sperm quality in fish: implications for cryobanking Programme National Programme
Instrument (FP6)
Contact Type (FP7)
Strand (Interreg)
NA
Theme (FP7)
Activity Area (FP6)
Regional Area (Interreg)
Action (COST)
NA
Specific Programme (FP7)
NA
Funding source National Coordinator Elsa Alexandra Martins / Silva Cabrita Coordinator email elsa.cabrita@icman.csic.es
Coordinator institution
CCMAR - Center of Marine Sciences (Portugal)
Institutions involved
FCT - Foundation for Science and Technology (Portugal) ,
Start year 2008 End year 2011 Funding (€) € 189,775 Website https://www.fct.pt/apoios/projectos/consulta/vglobal_projecto?idProjecto=64533&idElemConcurso=860 Summary Cryopreservation is a technique of significant interest. Sperm cryobanking has been used in conservation programs, genetic improvement of wild and captive populations, and is used in the selection of specific fish strain lines of commercial and conservation interest. The project aims to develop and standardize protocols for the cryopreservation of sperm from two important commercial species: turbot (Scophthalmus maximus) and seabass (Dicentrarchus labrax). Sperm cryopreservation from these species would allow for the establishment a selective breeding program and, from a production point a view, the use of cryopreserved sperm would reduce the costs of maintaining male broodstock out off reproductive season and would guarantee the availability of sperm whenever needed. Standardization of protocols requires a thorough and exhaustive evaluation of sperm quality. Therefore, the development of tools to characterize sperm quality should be available for widespread use for quality control and for the validation of freezing protocols. Cryopreservation can damage cells, affecting plasma membrane, organelles (in particular the mitochondrion) and chromatin. Damage is caused either by mechanical forces due to ice crystal formation, both within the cells and in the external medium or due to osmotic stress to which cells are exposed during cryoprotectant exposure and subsequent freezing/thawing procedures. The project aims to resolve these problems with 5 main objective. The first objective is to determine sperm quality in turbot and seabass and to select a group of parameters that allow the characterization of sperm quality in these two species. This will allow for the establishment of cryopreservation protocols for each species where several cryoprotectants, additives and extender solutions will be checked for the protection of all spermatozoa functions. These objectives will be achieved following the development of tools to evaluate sperm quality after freezing, identifying the principal type of damage produced at membrane, mitochondrion and DNA level. The conservation of each cell structure as well as its functionality will depend on the viability of the cryopreservation protocol. Most of the damage observed in spermatozoa after freezing/thawing can compromise sperm fertilization ability; however there is some evidence that cryopreservation can produce sublethal damage, not compromising fertility but affecting embryonic development. Thus, the last objective of the project will be the development of molecular tools for the identification and quantification of DNA damage in cryopreserved samples. In this objective, separated in two tasks, we intend to characterize the type of DNA damage using several techniques (Comet assay, TUNEL, SCSA, QPCR) and correlate with fertilizing and embryo development ability of DNA-damaged spermatozoa. The members of the team belong to two groups from CCMAR- the Aquaculture group, with significant experience in broodstock management and fish rearing, cryopreservation techniques and analysis of cryodamage and the Molecular Biology group with huge expertise in molecular tools. The joint efforts of both groups will allow the development of the project.
Keywords
Turbot;
Fish reproduction;
Seabass;
Flatfish;
Restocking;
Genetic;
Fish;
Marine Region
76
Not associated to marine areas
0
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