Intravenous microinjections of zebrafish larvae to study acute kidney injury

J Vis Exp. 2010 Aug 4:(42):2079. doi: 10.3791/2079.

Abstract

In this video article we describe a zebrafish model of AKI using gentamicin as the nephrotoxicant. The technique consists of intravenous microinjections on 2 dpf zebrafish. This technique represents an efficient and rapid method to deliver soluble substances into the bloodstream of zebrafish larvae, allowing for the injection of 15-20 fish per hour. In addition to AKI studies, this microinjection technique can also be used for other types of experimental studies such as angiography. We provide a detailed protocol of the technique from equipment required to visual measures of decreased kidney function. In addition, we also demonstrate the process of fixation, whole mount immunohistochemistry with a kidney tubule marker, plastic embedding and sectioning of the larval zebrafish. We demonstrate that zebrafish larvae injected with gentamicin show morphological features consistent with AKI: edema, loss of cell polarity in proximal tubular epithelial cells, and morphological disruption of the tubule.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Acute Kidney Injury*
  • Animals
  • Disease Models, Animal*
  • Female
  • Gentamicins / administration & dosage
  • Injections, Intravenous
  • Larva
  • Male
  • Microinjections / methods*
  • Zebrafish

Substances

  • Gentamicins