Effects of Cyclin Dependent Kinase 9 inhibition on zebrafish larvae

Cell Cycle. 2016 Nov 16;15(22):3060-3069. doi: 10.1080/15384101.2016.1231283. Epub 2016 Oct 7.

Abstract

CDK9 is a known regulator of cellular transcription, growth and proliferation. Small molecule inhibitors are currently being developed and assessed in clinical trials as anti-cancer drugs. The zebrafish embryo provides an ideal model to explore the effects of CDK9 inhibition in-vivo. This has not been adequately explored previously at the level of a whole organism. We have compared and contrasted the effects of pharmacological and molecular inhibition of CDK9 on somatic growth, apoptosis and cellular proliferation in zebrafish larvae between 0 to 120 hours post fertilisation (hpf) using flavopiridol, a selective CDK9 antagonist, and CDK9-targeting morpholino. We demonstrate that the inhibition of CDK9 diminishes cellular proliferation and increases apoptosis. Subsequently, it affects somatic growth and development of a number of key embryonic structures including the brain, heart, eye and blood vessels. For the first time, we have localized CDK9 at a subcellular level in whole-mounted larvae. This works shows, at a high-throughput level, that CDK9 clearly plays a fundamental role in early cellular growth and proliferation.

Keywords: CDK9; flavopiridol; morpholino; pharmacology; phenotype; zebrafish.

MeSH terms

  • Animals
  • Bromodeoxyuridine / metabolism
  • Cell Death / drug effects
  • Cell Proliferation / drug effects
  • Cyclin-Dependent Kinase 9 / antagonists & inhibitors*
  • Cyclin-Dependent Kinase 9 / metabolism
  • Embryo, Nonmammalian / drug effects
  • Embryo, Nonmammalian / metabolism
  • Flavonoids / pharmacology
  • Immunohistochemistry
  • Kaplan-Meier Estimate
  • Larva / drug effects
  • Morpholinos / pharmacology
  • Phenotype
  • Piperidines / pharmacology
  • Protein Kinase Inhibitors / pharmacology
  • Survival Analysis
  • Zebrafish / embryology
  • Zebrafish / growth & development*
  • Zebrafish / metabolism*

Substances

  • Flavonoids
  • Morpholinos
  • Piperidines
  • Protein Kinase Inhibitors
  • alvocidib
  • Cyclin-Dependent Kinase 9
  • Bromodeoxyuridine