CRISPR-Cas-Induced Mutants Identify a Requirement for dSTIM in Larval Dopaminergic Cells of Drosophila melanogaster

G3 (Bethesda). 2017 Mar 10;7(3):923-933. doi: 10.1534/g3.116.038539.

Abstract

Molecular components of store-operated calcium entry have been identified in the recent past and consist of the endoplasmic reticulum (ER) membrane-resident calcium sensor STIM and the plasma membrane-localized calcium channel Orai. The physiological function of STIM and Orai is best defined in vertebrate immune cells. However, genetic studies with RNAi strains in Drosophila suggest a role in neuronal development and function. We generated a CRISPR-Cas-mediated deletion for the gene encoding STIM in Drosophila (dSTIM), which we demonstrate is larval lethal. To study STIM function in neurons, we merged the CRISPR-Cas9 method with the UAS-GAL4 system to generate either tissue- or cell type-specific inducible STIM knockouts (KOs). Our data identify an essential role for STIM in larval dopaminergic cells. The molecular basis for this cell-specific requirement needs further investigation.

Keywords: Orai; SOCE; hypoderm; null allele; tyrosine hydroxylase.

Publication types

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

MeSH terms

  • Animals
  • Body Size
  • CRISPR-Cas Systems / genetics*
  • Dopaminergic Neurons / cytology*
  • Dopaminergic Neurons / metabolism*
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / anatomy & histology
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / genetics*
  • Gene Knockout Techniques
  • Homozygote
  • Larva / metabolism
  • Mutation / genetics*
  • Neurons / metabolism
  • Organ Specificity / genetics

Substances

  • Drosophila Proteins