Tissue-Specific IAG Interactome in Cherax quadricarinatus Gonads

Area of Science:

• * Molecular endocrinology
• * Crustacean biology
• * Reproductive biology

Background:

• * The insulin-like androgenic gland (IAG) hormone is crucial for sex differentiation in decapod crustaceans.
• * The molecular mechanisms underlying IAG's gonadal functions are not well understood.
• * Characterizing IAG interactions is key to understanding crustacean reproductive development.

Purpose of the Study:

• * To identify proteins interacting with IAG in the gonads of Cherax quadricarinatus.
• * To elucidate the tissue-specific molecular networks of IAG in male and female reproductive tissues.
• * To explore the functional implications of IAG interactions in sex differentiation and maturation.

Main Methods:

• * Protein pull-down assays were used to capture IAG-interacting proteins from androgenic gland (AG), testis, and ovary.
• * Gene Ontology (GO) and KEGG pathway enrichment analyses were performed.
• * High-confidence interactors were identified using structure prediction and reliability thresholds.
• * AlphaFold3 was used to predict binding sites and energies for key candidates.

Main Results:

• * 279 candidate IAG-interacting proteins were identified, with the AG showing the most interactors, enriched in metabolic pathways.
• * 32 high-confidence interactors were determined.
• * Interactions with UBA1, HSP, and HKMT in male tissues suggest potential epigenetic roles for IAG.
• * Interactions with DICER1 and CK2 in the ovary suggest post-transcriptional and post-translational inhibition of female maturation.

Conclusions:

• * The IAG interactome is tissue-specific, highlighting complex regulatory networks in crustacean gonads.
• * IAG plays a dual role: regulating metabolism in the AG and modulating post-transcriptional/post-translational events in the ovary.
• * Findings offer new insights into crustacean sex development and potential targets for sex-control strategies.