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.

类胰岛素雄激素基因（IAG）主要在雄激素腺（AG）中表达，在控制十足类雄性性别分化和维持雄性第二性征方面发挥着至关重要的作用。在本研究中，我们研究了雄性克氏原螯虾的mRNA和microRNA表达谱，以了解注射基于IAG设计的高效siRNA（GsiRNA）后IAG的转录组调控机制。结果显示，多个差异表达基因富集于生殖相关通路，如wnt信号通路、MAPK信号通路、GnRH信号通路。在睾丸（Te）中，注射GsiRNA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，大脑（Br）和腹神经索（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，Br/AN 中五种神经肽基因（甲壳类高血糖激素、色素分散激素、红色素浓缩激素前体、corazonin和促性腺激素释放激素 II 受体亚型 X1）的表达发生显着变化。此外，Te/AG 中高表达的三种卵巢相关 miRNA（miR-263a、miR-263b、miR-133）在注射 GsiRNA 后表现出显着上调。此外，GsiRNA的长期干扰被发现会抑制青少年阶段男性外性征的发育，并延迟成年阶段的发育。这项研究为克氏对虾IAG的分子调控机制和功能提供了有价值的见解。