Dissection of central clock function in Drosophila through cell-specific CRISPR-mediated clock gene disruption

Elife. 2019 Oct 15:8:e48308. doi: 10.7554/eLife.48308.

Abstract

In Drosophila, ~150 neurons expressing molecular clock proteins regulate circadian behavior. Sixteen of these neurons secrete the neuropeptide Pdf and have been called 'master pacemakers' because they are essential for circadian rhythms. A subset of Pdf+ neurons (the morning oscillator) regulates morning activity and communicates with other non-Pdf+ neurons, including a subset called the evening oscillator. It has been assumed that the molecular clock in Pdf+ neurons is required for these functions. To test this, we developed and validated Gal4-UAS based CRISPR tools for cell-specific disruption of key molecular clock components, period and timeless. While loss of the molecular clock in both the morning and evening oscillators eliminates circadian locomotor activity, the molecular clock in either oscillator alone is sufficient to rescue circadian locomotor activity in the absence of the other. This suggests that clock neurons do not act in a hierarchy but as a distributed network to regulate circadian activity.

Keywords: CRISPR/Cas9; D. melanogaster; GAL4/UAS; circadian rhythm; genetics; genomics; neuroscience.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology
  • Brain / metabolism
  • Brain / radiation effects
  • CRISPR-Cas Systems
  • Cell Communication
  • Cell Lineage / genetics
  • Circadian Clocks / drug effects
  • Circadian Clocks / genetics*
  • Circadian Rhythm / drug effects
  • Circadian Rhythm / genetics*
  • Darkness
  • Drosophila Proteins / deficiency
  • Drosophila Proteins / genetics*
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / metabolism
  • Drosophila melanogaster / radiation effects
  • Feedback, Physiological
  • Gene Editing
  • Gene Expression Regulation
  • Light Signal Transduction / genetics
  • Locomotion / genetics
  • Locomotion / radiation effects
  • Nerve Net / metabolism
  • Nerve Net / radiation effects
  • Neurons / cytology
  • Neurons / metabolism*
  • Neurons / radiation effects
  • Neuropeptides / deficiency
  • Neuropeptides / genetics*
  • Period Circadian Proteins / deficiency
  • Period Circadian Proteins / genetics*
  • Transcription Factors / deficiency
  • Transcription Factors / genetics

Substances

  • Drosophila Proteins
  • GAL4 protein, Drosophila
  • Neuropeptides
  • PER protein, Drosophila
  • Period Circadian Proteins
  • Transcription Factors
  • pdf protein, Drosophila
  • tim protein, Drosophila