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

Intravenous microinjections of zebrafish larvae to study acute kidney injury.

Author information

1
Department of Developmental Biology, University of Pittsburgh, PA, USA.

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.

PMID:
20729805
PMCID:
PMC3156013
DOI:
10.3791/2079
[Indexed for MEDLINE]
Free PMC Article

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