Induction of germ cell-deficiency in grass puffer by dead end 1 gene knockdown for use as a recipient in surrogate production of tiger puffer
Induction of germ cell-deficiency in grass puffer by dead end 1 gene knockdown for use as a recipient in surrogate production of tiger puffer
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DOI：
10.1016/j.aquaculture.2020.735385
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发表时间：
2020-09
期刊：
影响因子：
4.5
通讯作者：
H. Yoshikawa;Yasuko Ino;K. Kishimoto;Hayato Koyakumaru;Taiju Saito;M. Kinoshita;Y. Yoshiura
中科院分区：
文献类型：
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作者：
H. Yoshikawa;Yasuko Ino;K. Kishimoto;Hayato Koyakumaru;Taiju Saito;M. Kinoshita;Y. Yoshiura
代理亲鱼技术通过将生殖细胞移植到代理鱼中来产生供体衍生的配子。该技术是改善具有理想遗传性状的水产养殖品种的建立和管理的一种有前景的方法。虎河豚（Takifugu rubripes）是日本最受欢迎的水产养殖鱼类之一。然而，亲鱼体型较大，需要很长时间才能达到性成熟。目前的研究在草鲀（T. alboplumbeus）中建立了一种利用基因敲除的绝育技术，草鲀体型较小，成熟时间是虎鲀的一半，并评估了使用生殖细胞缺陷的草鲀作为受体来有效产生供体来源的虎鲀配子的可能性。在草鲀中鉴定出了具有两个转录变异体的Dead end 1。将针对转录变体的反义吗啉寡核苷酸显微注射到受精卵中产生的畸形体显示出生殖细胞缺陷。生殖细胞缺陷的突变体在形态和基因表达模式方面表现出性别二态性。从虎河豚睾丸中制备的睾丸生殖细胞被腹腔内移植到吗啡孵化的幼体中。这些受体正常成熟，并产生与内源配子分开的功能性供体衍生配子。因此，使用仅产生供体衍生配子的生殖细胞缺陷受体可以提高代孕生产的效率，并可能加速虎河鲀水产养殖的育种过程。
The surrogate broodstock technique produces donor-derived gametes via transplantation of germ cells into surrogate fish. The technique is a promising approach for improving the establishment and management of aquaculture strains with desirable genetic traits. The tiger puffer (Takifugu rubripes) is one of the most popular aquaculture fishes in Japan. However, brood stocks have large body sizes and require a long time to reach sexual maturity. The current study established a sterilization technique using gene knockdown in the grass puffer (T. alboplumbeus), which has a small body size and matures in half the time taken by the tiger puffer, and assessed the possibility of using germ cell-deficient grass puffers as recipients to efficiently produce donor-derived tiger puffer gametes.Dead end 1,which has two transcribed variants, was identified in the grass puffer. Morphants resulting from the microinjection of antisense morpholino oligonucleotides against the transcribed variants, into fertilized eggs, showed germ cell deficiency. Germ cell-deficient morphants exhibited sexual dimorphism with respect to their morphological and gene expression patterns. Testicular germ cells prepared from the testes of tiger puffer were intraperitoneally transplanted into morphant hatchlings. These recipients matured normally and produced functional donor-derived gametes separately from endogenous gametes. Thus, the use of germ cell-deficient recipients that produce only donor-derived gametes improves the efficiency of surrogate production and may accelerate the breeding process in tiger puffer aquaculture.