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 and Rural Affair, Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
School of Life Sciences, Huzhou University, Huzhou, 313000, Zhejiang, China.
Mar Biotechnol (NY). 2023 Dec;25(6):983-996. doi: 10.1007/s10126-023-10254-z. Epub 2023 Oct 13.
The insulin-like androgenic gland hormone gene (IAG), primarily expressed in the androgenic gland (AG), plays a crucial role in controlling male sex differentiation and maintaining male secondary sexual characteristics in decapods. In this study, we investigated the mRNA and microRNA expression profiles of male Procambarus clarkii to understand the transcriptomic regulatory mechanism of IAG after the injection of an efficient siRNA (GsiRNA) designed based on IAG. The results revealed that several differentially expressed genes were enriched in reproduction-related pathways, such as the wnt signaling pathway, MAPK signaling pathway, and GnRH signaling pathway. In the testis (Te), the injection of GsiRNA led to the up-regulation of many ovary-related genes and down-regulation of testis-related genes. Moreover, the brain (Br) and abdominal nerve cord (AN) appeared to be involved in the regulation of IAG, with numerous differentially expressed genes found in Br and AN. Notably, the expression of five neuropeptide genes, Crustacean hyperglycemic hormone, pigment-dispersing hormone, red pigment concentrating hormone precursor, corazonin, and gonadotropin-releasing hormone II receptor isoform X1 in Br/AN, was significantly changed. Additionally, three ovary-related miRNAs (miR-263a, miR-263b, miR-133) highly expressed in Te/AG showed significant up-regulation after GsiRNA injection. Furthermore, the long-term interference of GsiRNA was found to inhibit the development of male external sexual characteristics during the juvenile stage and delay it during the adult stage. This research provides valuable insights into the molecular regulatory mechanism and function of IAG in P. clarkii.
胰岛素样雄激素腺激素基因（IAG）主要在雄激素腺（AG）中表达，在控制十足目动物雄性性别分化和维持雄性第二性征方面发挥着关键作用。在这项研究中，我们研究了雄性克氏原螯虾的 mRNA 和 microRNA 表达谱，以了解基于 IAG 设计的高效 siRNA（GsiRNA）注射后 IAG 的转录调控机制。结果表明，几个差异表达基因富集在与生殖相关的途径中，如 wnt 信号通路、MAPK 信号通路和 GnRH 信号通路。在睾丸（Te）中，GsiRNA 的注射导致许多卵巢相关基因的上调和睾丸相关基因的下调。此外，脑（Br）和腹部神经索（AN）似乎参与了 IAG 的调节，在 Br 和 AN 中发现了许多差异表达的基因。值得注意的是，五种神经肽基因，甲壳动物高血糖素、色素分散激素、红色素浓缩激素前体、心皮质素和促性腺激素释放激素 II 受体 X1 的表达在 Br/AN 中发生了显著变化。此外，在 Te/AG 中高度表达的三种卵巢相关 miRNA（miR-263a、miR-263b、miR-133）在 GsiRNA 注射后表现出明显的上调。此外，发现 GsiRNA 的长期干扰会抑制雄性外部生殖特征在幼体阶段的发育，并在成年阶段延迟。这项研究为 IAG 在 P. clarkii 中的分子调控机制和功能提供了有价值的见解。
