College of Fisheries, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education/Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, China.
College of Natural and Applied Sciences, University of Guam, Mangilao, GU, USA.
Mar Biotechnol (NY). 2020 Jun;22(3):456-466. doi: 10.1007/s10126-020-09965-4. Epub 2020 Apr 27.
The insulin-like androgenic gland (IAG) gene is well known in male crustacean, and it is a key regulator in male sexual differentiation and maintaining the male sexual characteristic. The neo-female can be produced by silencing the MrIAG (Macrobrachium rosenbergii Insulin-like Androgenic Gland) in male Macrobrachium rosenbergii. This is the first time to use siRNA approach to silenced MrIAG in male M. rosenbergii. In the current study, the optimal injection dosage to achieve sex reversal is 0.5 μg/g body weight. After MrIAG silencing, the expression level of Dmrt11e, Dmrt99b, MRPINK, Mrr, Sxl1, and Sxl2 decreased significantly. As their long-term silencing effect of MrIAG, the dsRNA and siRNA approaches produce three and two individual neo-females, respectively. The neo-female has a wider brood chamber, ovipositing setae, and ovigerous setae, which is resembled normal female. After a long-term silencing with siRNA, most of the germ cells were arrested in spermatocytes stage, but the spermatocytes in control can further developed into spermatozoon. The seminiferous tubules are loosely arranged and the spermatocytes are more than spermatozoon in the 0.5 μg/g body weight treatment dose. This current study suggests a new path to obtain neo-females through siRNA silencing.
胰岛素样雄激素腺（IAG）基因在雄性甲壳动物中广为人知，它是雄性性分化和维持雄性性特征的关键调节剂。通过沉默雄性罗氏沼虾（Macrobrachium rosenbergii Insulin-like Androgenic Gland）的 MrIAG，可以产生新雌性。这是首次使用 siRNA 方法沉默雄性 M. rosenbergii 的 MrIAG。在本研究中，实现性反转的最佳注射剂量为 0.5μg/g 体重。在 MrIAG 沉默后，Dmrt11e、Dmrt99b、MRPINK、Mrr、Sxl1 和 Sxl2 的表达水平显著下降。作为 MrIAG 的长期沉默效果，dsRNA 和 siRNA 方法分别产生了三个和两个新的雌性个体。新雌性具有更宽的生殖腔、产卵刚毛和产卵栉状刚毛，类似于正常雌性。通过 siRNA 长期沉默后，大多数生殖细胞停滞在精母细胞阶段，但对照组中的精母细胞可以进一步发育成精子。精小管排列松散，在 0.5μg/g 体重处理剂量下，精母细胞多于精子。本研究为通过 siRNA 沉默获得新雌性提供了一条新途径。
