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[Display omitted] • IGFBP-rp expression was assessed in the swimming crab Portunus trituberculatus. • PtIGFBP-rp and PtIAG share similarity in expression patterns during AG development. • PtIGFBP-rp may be involved in IAG signaling network by direct or indirect regulation of PtIAG gene. The insulin-like growth factor/insulin-like polypeptide (IGF/ILP) signaling is vital for growth, physiological metabolism, development, and reproduction. Insulin-like growth factor-binding protein (IGFBP) is involved in the insulin signaling pathway in both vertebrates and invertebrates and is critical for various physiology functions. Herein, we cloned and characterized the full-length cDNA of IGFBP-rp in the swimming crab, Portunus trituberculatus (PtIGFBP-rp). The deduced amino acid sequence of PtIGFBP-rp was found to contain three key domains (insulin-like binding (IB) domain, the kazale-type serine protease inhibitor (KAZAL) domain, and the immunoglobulin-like C2 (IGc2) domain). Results showed that PtIGFBP-rp shared the same expression pattern as P. trituberculatus insulin androgenic gland hormone (PtIAG) transcripts during the embryonic larval, juvenile crab stage and the androgenic gland (AG) developmental cycle. Moreover, PtIGFBP-rp transcripts were also present in high abundance in hepatopancreas, muscle, and androgenic glands. The regulatory relationship between PtIGFBP-rp and PtIAG was investigated by RNA interference and co-localization assays, which showed a co-localization relationship and feedback regulation between them. Bilateral eye stalk ablation (ESA) increased the expression of PtIGFBP-rp in the AG at 7 d after surgery. These results demonstrate the involvement of PtIGFBP-rp in the signaling regulatory network of IAG in P. trituberculatus. [ABSTRACT FROM AUTHOR]

Abstract Similar to the role of juvenile hormone (JH) in insects, methyl farnesoate (MF), the unepoxidized form of JH III, regulates many developmental processes in crustaceans, such as molting and reproduction. We have previously showed that the JH receptor, Methoprene-tolerant (Met), which is also a candidate receptor for MF, might be involved in the MF-mediated vitellogenesis in the swimming crab Portunus trituberculatus. In this study, the role of Kruppel homolog 1 (Kr-h1), a transcription factor downstream Met in JH signaling, was further investigated. The deduced protein of Pt-Kr-h1 contained seven repeats of zinc finger motifs, similar to Kr-h1s from other crustacean species, but differing from the eight zinc finger motifs found in insect Kr-h1s. MF treatment in vitro induced the expression of Pt-Kr-h1 in hepatopancreas but not ovary, which is similar to the MF-responsive pattern of Pt-Met as previously reported. Moreover, the expression of Pt-Kr-h1 decreased significantly after treating with Pt-Met dsRNA, strongly indicating that the Pt-Kr-h1 might be involved in the Met-mediated MF signaling pathway. RNAi of Pt-Met and Pt-Kr-h1 both led to a decrease in vitellogenin (Vg) expression, and the reduction cannot be rescued by adding MF, suggesting the regulation of vitellogenesis by MF may act through Met and Kr-h1. These results would help to enhance the current understanding of the regulatory mechanism of MF signaling, and provide a vital resource for further research into the evolution of hormonal pathways in arthropods. Highlights • Seven zinc finger motifs may be a common characteristic for crustacean Kr-h1s. • Kr-h1 is a MF-induced gene in P. trituberculatus. • MF may stimulate the exogenous vitellogenesis through Met and Kr-h1. [ABSTRACT FROM AUTHOR]