Majhi Sullip Kumar, Rasal Avinash Rambhau, Kushwaha Basdeo, Raizada Sudhir
ICAR-National Bureau of Fish Genetic Resources, Canal Ring Road, Dilkhusa P.O., Lucknow 226002, Uttar Pradesh, India.
ICAR-Central Institute of Freshwater Aquaculture, Kausalyanga, Bhubaneswar-751002, Odisha, India.
Anim Reprod Sci. 2017 Aug;183:77-85. doi: 10.1016/j.anireprosci.2017.05.015. Epub 2017 Jun 3.
Several options have been proposed for eradication of germ cells (GCs) in mammals such as treatment with cytotoxic drugs, irradiation, cold ischemia and hyperthermic treatment. Some of these methods have been also tried in fish but conditions for complete sterilisation of gonads have not been established. Here, we report the production of sterile adult common carp Cyprinus carpio in 10 weeks by the heat and chemical treatments. The cytotoxic drug busulfan (40mg/kg) was intraperitoneally injected into the animals at 2-week intervals (5 doses in total), and they were maintained in water at 38°C between Weeks 1 and 10. The effectiveness of the treatments was assessed using gonadal index, histology, and vasa gene expression. At the end of Week 10, very severe gonadal degeneration was observed in fish treated with the heat-chemical combination, and 100% of male and female fish were devoid of endogenous GCs. The average levels of vasa transcript were 0.01±0.005 and 0.02±0.016 for males and females, respectively. By contrast, high temperature alone caused minor gonadal degeneration and the gene transcript were 0.59±0.131 for male and 0.62±0.13 for female. In Week 20, after the recovery period of 10 weeks at 25°C, the gonadal germ cell did not recover from the sterile condition in any of the sampled individuals. The change in colouration of gonads was an additional useful index of the degree of gonadal sterility.
为根除哺乳动物的生殖细胞(GCs)已提出了几种方法,如使用细胞毒性药物治疗、辐射、冷缺血和高温处理。其中一些方法也已在鱼类中尝试,但尚未确定使性腺完全绝育的条件。在此,我们报告通过热化学处理在10周内培育出不育成年鲤鱼(Cyprinus carpio)。将细胞毒性药物白消安(40mg/kg)以2周的间隔腹腔注射到动物体内(共5剂),并在第1周和第10周期间将它们饲养在38°C的水中。使用性腺指数、组织学和血管生成素基因表达评估处理的有效性。在第10周结束时,观察到经热化学联合处理的鱼性腺发生非常严重的退化,100%的雄鱼和雌鱼均无内源性生殖细胞。雄鱼和雌鱼血管生成素转录本的平均水平分别为0.01±0.005和0.02±0.016。相比之下,单独高温仅导致轻微的性腺退化,雄鱼和雌鱼的基因转录本分别为0.59±0.131和0.62±0.13。在第20周,在25°C下恢复10周后,任何采样个体的性腺生殖细胞均未从不育状态恢复。性腺颜色的变化是性腺不育程度的另一个有用指标。