Short neuropeptide F receptors regulate feeding and reproduction in Hyphantria cunea

Neuropeptides are endogenous active compounds in the nervous system involved in regulating physiological processes such as growth, development, reproduction, and dynamic balance. In this study, two short neuropeptide F receptors (sNPFRs) were identified in Hyphantria cunea and exhibit different expression patterns among various developmental stages and tissues. The expression levels of sNPFR-A10 and sNPFR-A11 reach peak during the 6th and 1st instar larvae stages, respectively. Both genes are highly expressed in the midgut, Malpighian tubules, and testis. Silencing gene expressions by using small interference RNAs in fifth instar larvae led to lower food intake, decreased growth rate, and reduced glucose and triglyceride content compared to the controls. Furthermore, silencing of sNPFRs in the pupal stage decreased egg production and hatchability, altered the morphology of eukaryotic sperm tracts, and impaired fertility by disrupting eupyrene sperm formation and motility. Molecular docking showed that sNPFRs were activated by the mature peptide sNPF using AlphaFold analysis. These findings demonstrate that sNPFRs are involved in the regulation of reproductive processes and are part of a larger neuropeptide network that integrates feeding, growth and development, and reproductive physiology in H. cunea, highlighting their potential as targets for control strategies.