Gut single-nucleus RNA sequencing reveals that NPF receptor regulates feeding behavior through mitochondrial energy...

The insect gastrointestinal tract not only facilitates nutrient digestion and absorption but also serves as an endocrine regulatory center that integrates peripheral and central cues to feeding behavior. Neuropeptide signaling plays a key role in this process, coordinating appetite, digestion, and metabolic homeostasis. Here, we investigated the functional role of the neuropeptide F receptor (NPFR) in feeding regulation and intestinal energy metabolism in Spodoptera frugiperda. Quantitative RT-PCR and immunohistochemistry revealed that both NPF and NPFR are abundantly expressed in larval gut tissues, suggesting their involvement in local feeding control. Functional assays demonstrated that NPFR knockout larvae (NPFR^-/-) exhibited significantly reduced food intake and body weight compared with wild-type (WT) individuals. Single-nucleus RNA sequencing (snRNA-seq) analysis further revealed that NPFR^-/- enteroendocrine cells displayed pronounced downregulation of oxidative phosphorylation (OXPHOS) genes, including COX1, COX2, ATP6, and CYTB, indicating mitochondrial dysfunction and impaired energy metabolism. Together, these results establish NPFR as a key regulator linking gut-derived neuropeptide signaling to metabolic energy balance and feeding behavior. This study provides the first integrative evidence connecting neuropeptide signaling with intestinal mitochondrial function in a major Lepidopteran pest, offering new molecular insights into insect adaptation and identifying NPFR as a promising target for environmentally sustainable pest management strategies.