4.4 Article

Molecular cloning, characterization and functional analysis of two neuropeptide F genes from the oriental river prawn (Macrobrachium nipponense)

Publisher

ELSEVIER SCIENCE INC
DOI: 10.1016/j.cbpa.2020.110844

Keywords

Macrobrachium nipponense; Neuropeptide F; Ovary maturation; mRNA expression; RNA interference

Funding

  1. National Key R&D Progrom of China [2018YFD0901303]
  2. earmarked fund for Jiangsu Agricultural Industry Technology System
  3. Central Publicinterest Scientific Institution Basal Research Fund CAFS [2019JBFM04, 2020TD36]
  4. New cultivar breeding Major Project of Jiangsu province [PZCZ201745]
  5. China Agriculture Research System-48 (CARS-48)

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In invertebrates, two neuropeptide F genes, Mn-NPF1 and Mn-NPF2, were identified in Macrobrachium nipponense, with their expression correlated with ovarian maturation. Mn-NPF1 was positively correlated, while Mn-NPF2 had opposing expression patterns. Both neuropeptides were found to be involved in the seasonal regulation of reproduction, with Mn-NPF2 playing a significant role in inhibiting ovarian maturation.
In invertebrates, neuropeptide F (NPF) has many regulatory functions, similar to NPY, its homologous peptide. In this study, two neuropeptide F genes were identified in Macrobrachium nipponense: Mn-NPF1 and Mn-NPF2. Mn-NPF2 shared the same amino acid sequence with Mn-NPF1, except for a 37 amino acid insert in the middle of the NPF region. The quantitative-PCR (qPCR) results indicated that Mn-NPF1 expression was positively correlated with ovarian maturation, whereas Mn-NPF2 had opposing expression patterns. Both Mn-NPFs were poorly expressed at early embryonic stages, but enhanced expression levels were observed up to day 10 after hatching, when the gonads began to differentiate. Ovary in situ hybridization (ISH) analyses showed that both Mn-NPFs were present at all stages, but were differentially localized to distinct compartments. Temperature gradient studies showed that both Mn-NPFs were implicated in the seasonal regulation of reproduction. A double-stranded (ds) RNA-Mn-NPF2 injection led to a significant 38.5% increase in the vitellogenin (VG) transcript (P < 0.05). These results demonstrated that Mn-NPF2 plays an important role in inhibiting ovarian maturation.

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