標題：Sysmex Partec 倍性分析儀三倍體斑馬魚的生產(chǎn)與代孕繁殖應用
Abstract:
We report for first time comparison of two approaches for zebrafish triploid production using cold shock and heat shock treatment. Subsequently, produced triploid zebrafish were used as a recipients for intraperitoneal transplantation of ovarian and testicular cells originating from vas:EGFP strain in order to verify their suitability for surrogate reproduction. Heat shock treatment was far more effective evaluated as success rate of triploid production and viability in comparison to cold shock. Triploids were produced with up to 100% efficiency in particular females. As expected, all triploids were males. Subsequently, germ cells transplantation revealed that triploids are suitable surrogate hosts. Production of donor-derived sperm was achieved in 23% and 16% of triploids transplanted by testicular and ovarian cells, respectively. Success of the transplantation was confirmed by positive GFP signal detected in gonads of dissected fish and stripped sperm. Germline transmission was confirmed by fertilization tests followed by PCR analysis of embryos. Reproductive success of germline chimera triploids evaluated as fertilization rate and progeny development was comparable to control groups.
Keywords: zebrafish, chromosome manipulation, triploid, germ cell transplantation, surrogate
reproduction
摘要：
我們首次報道了采用冷休克與熱休克處理兩種方法生產(chǎn)斑馬魚三倍體的對比研究。隨后，將所獲三倍體斑馬魚作為受體，用于腹腔移植來自vas:EGFP品系的卵巢與睪丸細胞，以驗證其作為代孕載體的適用性。評估結(jié)果顯示，熱休克處理在三倍體生產(chǎn)成功率及存活率方面遠優(yōu)于冷休克處理。部分雌性個體的三倍體生產(chǎn)效率高達100%。不出所料，所有三倍體均為雄性。進一步的生殖細胞移植實驗表明，三倍體斑馬魚適合作為代孕載體。
在睪丸和卵巢細胞移植的三倍體中，分別有23%和16%實現(xiàn)了供體精子的產(chǎn)生。通過在解剖的魚和剝離的精子的性腺中檢測到陽性GFP信號，證實了移植的成功。通過受精試驗和胚胎PCR分析證實了種系傳播。通過受精率和后代發(fā)育評估，種系嵌合體三倍體的繁殖成功率與對照組相當。
關鍵詞：Sysmex Partec 倍性分析儀，斑馬魚、染色體操作、三倍體、生殖細胞移植、代孕
