PUBLICATION

Micromanipulation of gene expression in the adult zebrafish brain using cerebroventricular microinjection of morpholino oligonucleotides

Authors
Kizil, C., Iltzsche, A., Kaslin, J., and Brand, M.
ID
ZDB-PUB-130708-57
Date
2013
Source
Journal of visualized experiments : JoVE   (75): e50415 (Journal)
Registered Authors
Brand, Michael, Kaslin, Jan, Kizil, Caghan
Keywords
none
MeSH Terms
  • Animals
  • Gene Expression/drug effects*
  • Gene Knockdown Techniques/methods
  • Injections, Intraventricular
  • Microinjections
  • Morpholinos/administration & dosage*
  • Morpholinos/genetics
  • Oligonucleotides, Antisense/administration & dosage*
  • Oligonucleotides, Antisense/genetics
  • Telencephalon/cytology
  • Telencephalon/physiology*
  • Zebrafish
PubMed
23728426 Full text @ J. Vis. Exp.
Abstract

Manipulation of gene expression in tissues is required to perform functional studies. In this paper, we demonstrate the cerebroventricular microinjection (CVMI) technique as a means to modulate gene expression in the adult zebrafish brain. By using CVMI, substances can be administered into the cerebroventricular fluid and be thoroughly distributed along the rostrocaudal axis of the brain. We particularly focus on the use of antisense morpholino oligonucleotides, which are potent tools for knocking down gene expression in vivo. In our method, when applied, morpholino molecules are taken up by the cells lining the ventricular surface. These cells include the radial glial cells, which act as neurogenic progenitors. Therefore, knocking down gene expression in the radial glial cells is of utmost importance to analyze the widespread neurogenesis response in zebrafish, and also would provide insight into how vertebrates could sustain adult neurogenesis response. Such an understanding would also help the efforts for clinical applications in human neurodegenerative disorders and central nervous system regeneration. Thus, we present the cerebroventricular microinjection method as a quick and efficient way to alter gene expression and neurogenesis response in the adult zebrafish forebrain. We also provide troubleshooting tips and other useful information on how to carry out the CVMI procedure.

Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes