Enteroendocrine peptides regulate feeding behavior via controlling intestinal contraction of the silkworm Bombyx mori

PLoS One. 2019 Jul 1;14(7):e0219050. doi: 10.1371/journal.pone.0219050. eCollection 2019.

Abstract

Our previous study demonstrated that predominant feeding inhibitory effects were found in the crude extracts of foregut and midgut of the silkworm Bombyx mori larvae. To address the entero-intestinal control crucial for the regulation of insect feeding behavior, the present study identified and functionally characterized feeding inhibitory peptides from the midgut of B. mori larvae. Purification and structural analyses revealed that the predominant inhibitory factors in the crude extracts were allatotropin (AT) and GSRYamide after its C-terminal sequence. In situ hybridization revealed that AT and GSRYamide were expressed in enteroendocrine cells in the posterior and anterior midgut, respectively. Receptor screening using Ca2+-imaging technique showed that the B. mori neuropeptide G protein-coupled receptor (BNGR)-A19 and -A22 acted as GSRYamide receptors and BNGR-A5 acted as an additional AT receptor. Expression analyses of these receptors and the results of the peristaltic motion assay indicated that these peptides participated in the regulation of intestinal contraction. Exposure of pharynx and ileum to AT and GSRYamide inhibited spontaneous contraction in ad libitum-fed larvae, while exposure of pharynx to GSRYamide did not inhibit contraction in non-fed larvae, indicating that the feeding state changed their sensitivity to inhibitory peptides. These different responses corresponded to different expression levels of their receptors in the pharynx. In addition, injection of AT and GSRYamide decreased esophageal contraction frequencies in the melamine-treated transparent larvae. These findings strongly suggest that these peptides exert feeding inhibitory effects by modulating intestinal contraction in response to their feeding state transition, eventually causing feeding termination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx / cytology
  • Bombyx / genetics
  • Bombyx / physiology*
  • Enteroendocrine Cells / physiology
  • Feeding Behavior / physiology*
  • Genes, Insect
  • Insect Hormones / genetics
  • Insect Hormones / physiology
  • Insect Proteins / genetics
  • Insect Proteins / physiology
  • Intestines / cytology
  • Intestines / physiology
  • Larva / genetics
  • Larva / physiology
  • Models, Biological
  • Muscle Contraction / physiology
  • Neuropeptides / genetics
  • Neuropeptides / physiology
  • Oligopeptides / genetics
  • Oligopeptides / physiology
  • Phylogeny
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / physiology
  • Signal Transduction

Substances

  • Insect Hormones
  • Insect Proteins
  • Neuropeptides
  • Oligopeptides
  • Receptors, G-Protein-Coupled
  • allatotropin

Grants and funding

This study was partially supported by the Japan Society for the Promotion of Science (JSPS) (https://www.jsps.go.jp/) KAKENHI (Grant Number 16K15066) to SN and grants from the Slovak Research and Development Agency (SRDA) (http://www.apvv.sk/) (APVV-14-0556, APVV-16-0395 and VEGA 2/0080/18) and the European Regional Development Fund (ERDF) (http://ec.europa.eu/regional_policy/en/funding/erdf/) (ITMS: 26240220044) to DZ and LR. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.