Intravenous microinjections of zebrafish larvae to study acute kidney injury.
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.
Cianciolo Cosentino
Chiara
C
Department of Developmental Biology, University of Pittsburgh, PA, USA.
Roman
Beth L
BL
Drummond
Iain A
IA
Hukriede
Neil A
NA
D058186:Acute Kidney Injury; D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D005839:Gentamicins / Q000008:administration & dosage; D007275:Injections, Intravenous; D007814:Larva; D008297:Male; D008845:Microinjections / Q000379:methods*; D015027:Zebrafish
D005839:Gentamicins