Abstract

Diuretic hormones (DH) related to the vertebrate Corticotropin Releasing Factor (CRF) have been identified in diverse insect species. In the migratory locust, Locusta migratoria, the CRF-like DH (CRF/DH) is localized in the same neurosecretory cells as the Ovary Maturating Parsin (OMP), a neurohormone that stimulates oocyte growth, vitellogenesis and hemolymph ecdysteroid levels in adult female locusts. In this study, we investigated whether CRF-like DH can influence feeding and reproduction in the desert locust, Schistocerca gregaria. We identified two highly similar S. gregaria CRF-like DH precursor cDNAs, each of which also encodes an OMP isoform. Alignment with other insect CRF-like DH precursors shows relatively high conservation of the CRF/DH sequence while the precursor region corresponding to OMP is not well conserved. Quantitative real-time RT-PCR revealed that the precursor transcripts mainly occur in the central nervous system and their highest expression level was observed in the brain. Injection of locust CRF/DH caused a significantly reduced food intake, while RNAi knockdown stimulated food intake. Therefore, our data indicate that CRF-like DH induces satiety. Furthermore, injection of CRF/DH in adult females retarded oocyte growth and caused lower ecdysteroid titers in hemolymph and ovaries, while RNAi knockdown resulted in opposite effects. The observed effects of CRF/DH may be part of a wider repertoire of neurohormonal activities, constituting an integrating control system that affects food intake and excretion, as well as anabolic processes like oocyte growth and ecdysteroidogenesis, following a meal. Our discussion about the functional relationship between CRF/DH and OMP led to the hypothesis that OMP may possibly act as a monitoring peptide that can elicit negative feedback effects.

Highlights

  • Insect diuretic neuropeptides stimulate primary urine secretion in the Malpighian tubules, the excretory organs

  • When the Corticotropin Releasing-Factor (CRF)-like diuretic hormone (DH) and Ovary Maturating Parsin (OMP) sequences from L. migratoria and S. gregaria are compared (Figure 2B; for S. gregaria the isoform most resembling to the Locmi-OMP was taken), we find a 100% identity for both CRF/DH sequences, while the OMP sequences show about 85% identity with most observed mutations being conservative

  • S. gregaria OMP-DH precursor transcript sequences In the present paper, we describe the cloning of two highly similar OMP-DH-precursor cDNAs in Schistocerca gregaria

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Summary

Introduction

Insect diuretic neuropeptides stimulate primary urine secretion in the Malpighian tubules, the excretory organs. Locmi-OMP is a neurohormone of 65 amino acids (6.9 kDa), which stimulates oocyte maturation and vitellogenesis when daily injected in young adult female L. migratoria [19,20] These OMP injections seemed to accelerate the occurrence of circulating ecdysteroids in the hemolymph [20,21]. In the recently reported S. gregaria EST (‘‘Expressed Sequence Tags’’) database, the 59 regions of two OMP-encoding precursor transcripts, each encoding one of the two large Schgr-OMP isoforms, were found [24] By completing both sequences with 39 Rapid Amplification of cDNA Ends (39RAcE), we identified two full precursor sequences and discovered that each of these encode an Schgr-OMP isoform, and contain the sequence of the locust CRF-like diuretic hormone. We were especially interested in the possible effects on oocyte growth and ecdysteroid levels, since it previously was described that OMP injection affects these parameters

Materials and Methods
Quantitative real-time PCR
Results
Discussion
Full Text
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