CRISPR knockouts reveal an endogenous role for ancient neuropeptides in regulating developmental timing in a sea anemone

Elife. 2018 Sep 18:7:e39742. doi: 10.7554/eLife.39742.

Abstract

Neuropeptides are evolutionarily ancient peptide hormones of the nervous and neuroendocrine systems, and are thought to have regulated metamorphosis in early animal ancestors. In particular, the deeply conserved Wamide family of neuropeptides-shared across Bilateria (e.g. insects and worms) and its sister group Cnidaria (e.g. jellyfishes and corals)-has been implicated in mediating life-cycle transitions, yet their endogenous roles remain poorly understood. By using CRISPR-Cas9-mediated reverse genetics, we show that cnidarian Wamide-referred to as GLWamide-regulates the timing of life cycle transition in the sea anemone cnidarian Nematostella vectensis. We find that mutant planula larvae lacking GLWamides transform into morphologically normal polyps at a rate slower than that of the wildtype control larvae. Treatment of GLWamide null mutant larvae with synthetic GLWamide peptides is sufficient to restore a normal rate of metamorphosis. These results demonstrate that GLWamide plays a dispensable, modulatory role in accelerating metamorphosis in a sea anemone.

Keywords: Cnidaria; Nematostella; development; developmental biology; evolution; evolutionary biology; metamorphosis; neuropeptides.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • CRISPR-Associated Protein 9 / metabolism
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • DNA, Complementary / genetics
  • Embryo, Nonmammalian / metabolism
  • Gene Knockout Techniques*
  • Larva
  • Mutagenesis / genetics
  • Mutation / genetics
  • Neurons / metabolism
  • Neuropeptides / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sea Anemones / embryology*
  • Sea Anemones / metabolism*
  • Time Factors

Substances

  • DNA, Complementary
  • Neuropeptides
  • RNA, Messenger
  • CRISPR-Associated Protein 9

Grants and funding

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.