Production and use of triploid zebrafish for surrogate reproduction

Abstract

12We report for first time comparison of two approaches for zebrafish triploid production using cold 13 shock and heat shock treatment. Subsequently, produced triploid zebrafish were used as a recipients 14 for intraperitoneal transplantation of ovarian and testicular cells originating from vas:EGFP strain in 15 order to verify their suitability for surrogate reproduction. Heat shock treatment was far more 16 effective evaluated as success rate of triploid production and viability in comparison to cold shock. 17Triploids were produced with up to 100% efficiency in particular females. As expected, all triploids 18 were males. Subsequently, germ cells transplantation revealed that triploids are suitable surrogate 19 hosts. Production of donor-derived sperm was achieved in 23% and 16% of triploids transplanted 20 by testicular and ovarian cells, respectively. Success of the transplantation was confirmed by 21 positive GFP signal detected in gonads of dissected fish and stripped sperm. Germline transmission 22