Molecular Characterization, Signaling Activity, and Expression Profiles of a Novel Prolactin Splice Variant (PRL-S)...

Intense broodiness restricts egg production in Zhedong White Geese, with prolactin (PRL) being a key regulator. This study aimed to elucidate the role of PRL gene alternative splice variants in regulating broodiness in Zhedong White Geese. A total of 20 Zhedong White Geese at 400 days of age were selected and divided into two groups of 10 based on their physiological states: the laying period and the brooding period. Through bioinformatics and molecular biology approaches, PRL-L and PRL-S were identified and characterized. The results showed that PRL-S lacked a signal peptide and the first 57 amino acids at the N-terminus, leading to the disappearance of its first α-helix structure. Functional validation demonstrated that the recombinant PRL-S protein, prepared using a prokaryotic expression system, possessed physiological activity, including receptor binding and the activation of downstream signaling pathways. Furthermore, Real-time PCR and Western blot analyses revealed that the expression of both splice variants in the hypothalamic-pituitary-ovarian axis exhibited significant spatiotemporal specificity and was closely associated with reproductive states. This study revealed the molecular characteristics, in vitro functional activity, and expression patterns of PRL-S, providing new insights into the regulatory mechanisms of PRL.