Captive maturation studies in Penaeus monodon by GIH silencing using constitutively expressed long hairpin RNA
Captive maturation studies in Penaeus monodon by GIH silencing using constitutively expressed long hairpin RNA
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DOI：
10.1016/j.aquaculture.2015.06.036
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发表时间：
2015-11-01
期刊：
影响因子：
4.5
通讯作者：
Chaudhari, Aparna
中科院分区：
文献类型：
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作者：
Das, Rekha;Krishna, Gopal;Chaudhari, Aparna
斑节对虾的圈养成熟和产卵目前是通过单侧眼柄切除诱导的，这种方法可以去除眼柄分泌的性腺抑制激素(GIH)。然而，由于眼柄分泌的其他荷尔蒙的意外去除，单侧眼柄切除会给育雏鸟带来生理上的并发症。在这里，我们研究了长发夹RNA沉默GIH对斑节对虾生殖生理的影响。我们观察到雄激素(AGH)转录本在雄性中的表达增加了3-5倍，但对雌性的卵黄原蛋白的表达没有影响。然而，在去势的动物中，成熟指示基因的转录在两性中都出现了积极的影响，并超过了沉默处理的效率。在GIH沉默和去的雄性的组织切片中都有精子生成活跃的迹象，但在雌性的组织水平上没有变化。我们还观察到，在实验结束时，GIH沉默的动物的蜕皮率和相关死亡率显著更高。我们的结果表明，眼柄切除仍然是诱导斑节对虾成熟的最有效的技术。虽然GIH沉默可能是一种潜在的替代眼柄消融的方法，但通过使用组织特异性和/或可诱导的启动子来表达lhRNA，还需要进一步的研究来提高其效率。同样明显的是，雌性需要相当大的体型才能成功进行这些干预。相关的声明诱导圈养成熟一直是对虾产业面临的挑战。自然采集的亲鱼丰度低、价格高，常常阻碍种子生产。天然亲鱼的采集也存在WSSV感染的固有问题。此外，除非生殖周期关闭，否则这种高度优先的物种的遗传改良是困难的。由于所有这些原因，重要的是研究该物种的生殖生理和内分泌学，以及如何操纵它以诱导圈养成熟。本研究利用RNA干扰机制沉默性腺抑制激素基因，以观察其对成熟的影响。(C)2015爱思唯尔B.V.保留所有权利。
Captive maturation and spawning in Penaeus monodon are currently induced by unilateral eyestalk ablation, a method that removes the gonad-inhibiting hormone (GIH) secreted from the eyestalk. However, unilateral eyestalk ablation creates physiological complications in the brooder due to the unintended removal of other hormones secreted by the eyestalks. Here we studied the effect of gih silencing by long hairpin RNA on the reproductive physiology of P. monodon. We observed 3-5 fold enhanced expression of the androgenic gland hormone (AGH) transcript in males, but no effect on vitellogenin expression in females. In the destalked animals, however, positive effect on the maturation indicator transcripts was seen in both the sexes, and surpassed the efficiency of the silencing treatment. There were indications of active spermatogenesis in histological sections of both the gih silenced and destalked males, but no changes at histological level in the females. We also observed a significantly higher rate of molting and associated mortality in the gih silenced animals at the end of the experiment. Our results suggest that eyestalk ablation is still the most efficient technique to induce maturation in P. monodon. Although gih silencing could be a potential alternative to eyestalk ablation, further research is needed to enhance its efficiency over eyestalk ablation by using tissue specific and/or inducible promoter for lhRNA expression. It is also evident that females need to be fairly large in size for these interventions to succeed.Statement of relevanceInduction of captive maturation has been a challenge for shrimp industry. The seed production is often hampered by the low abundance and high price of naturally collected broodstock. Collection of natural brooders also has the intrinsic problem of WSSV infection. Also, genetic improvement of this highly preferred species is difficult unless the reproductive cycle is closed. For all these reasons it is important to study the reproductive physiology and endocrinology of this species and ways to manipulate it in order to induce captive maturation. The present study utilizes the RNA interference mechanism to silence the gonad-inhibiting hormone gene to see its effect on maturation. (C) 2015 Elsevier B.V. All rights reserved.