Generation of juvenile rainbow trout derived from cryopreserved whole ovaries by intraperitoneal transplantation of ovarian germ cells
Generation of juvenile rainbow trout derived from cryopreserved whole ovaries by intraperitoneal transplantation of ovarian germ cells
复制标题
DOI：
10.1016/j.bbrc.2016.08.156
复制
发表时间：
2016-09-23
影响因子：
3.1
通讯作者：
Yoshizaki, Goro
中科院分区：
文献类型：
--
作者：
Lee, Seungki;Katayama, Naoto;Yoshizaki, Goro
鱼类精子的冷冻保存为选择性育种和生物多样性保护提供了实际应用。然而，由于缺乏鱼卵和胚胎的冷冻保存方法，母系遗传的细胞质区室无法成功保存。我们之前开发了一种替代方法，通过将睾丸细胞移植到女性和男性受体中，从冷冻保存的整个睾丸中获取功能性卵子和精子。然而，如果目标鱼有卵巢，以前使用雄性生殖细胞的方法将无效。在这里，我们的目标是通过将卵巢生殖细胞移植到无菌孵化体的腹膜腔中，从冷冻保存的整个卵巢中产生功能性配子。通过测试几种不同的冷冻保护剂，优化了虹鳟鱼卵巢的冷冻保存条件（1.0 M DMSO、0.1 M 海藻糖和 10% 蛋黄）。来自解冻卵巢的卵巢生殖细胞被腹膜内移植到同种异体三倍体幼体中。移植的生殖细胞迁移并融入受体性腺，在那里进行配子发生。冷冻保存长达1185天后，卵巢生殖细胞的移植效率保持稳定。尽管所有未接受移植的三倍体受者均具有功能性不育，但 25 名女性受者中的 5 名和 25 名男性受者中的 7 名在移植后 2.5 年达到性成熟。对所得卵子和精子进行授精，产生了可存活的后代，这些后代表现出橙色体色、绿色荧光和染色体数量等供体特征。因此，该方法是保护濒危鱼类的突破性工具，对于冷冻保存雌鱼遗传资源至关重要。 (C) 2016 Elsevier Inc. 保留所有权利。
Cryopreservation of fish sperm offers the practical applications in the selective breeding and biodiversity conservation. However, because of the lack of cryopreservation methods for fish eggs and embryos, maternally inherited cytoplasmic compartments cannot be successfully preserved. We previously developed an alternative method to derive functional eggs and sperm from cryopreserved whole testis by transplanting testicular cells into female and male recipients. However, if target fish had ovaries, the previous method employing male-derived germ cells would be ineffective. Here, we aimed to generate functional gametes from cryopreserved whole ovaries by transplanting ovarian germ cells into peritoneal cavity of sterile hatchlings. Cryopreservation conditions for rainbow trout ovaries (1.0 M DMSO, 0.1 M trehalose, and 10% egg yolk) were optimized by testing several different cryoprotective agents. Ovarian germ cells from thawed ovaries were intraperitoneally transplanted into allogeneic triploid hatchlings. Transplanted germ cells migrated toward and were incorporated into recipient gonads, where they underwent gametogenesis. Transplantation efficiency of ovarian germ cells remained stable after cryopreservation period up to 1185 days. Although all triploid recipients that did not undergo transplantation were functionally sterile, 5 of 25 female recipients and 7 of 25 male recipients reached sexual maturity at 2.5 years post-transplantation. Inseminating the resultant eggs and sperm generated viable offspring displaying the donor characteristics of orange body color, green fluorescence, and chromosome numbers. This method is thus a breakthrough tool for the conservation of endangered fish species that are crucial to cryopreserve the genetic resources of female fish. (C) 2016 Elsevier Inc. All rights reserved.