Intravenous Microinjections of Zebrafish Larvae to Study Acute Kidney Injury |
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Authors: | Chiara Cianciolo Cosentino Beth L. Roman Iain A. Drummond Neil A. Hukriede |
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Affiliation: | Department of Developmental Biology, University of Pittsburgh;Department of Biological Sciences, University of Pittsburgh;Department of Medicine and Genetics, Harvard Medical School |
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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. |
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Keywords: | Developmental Biology Issue 42 intravenous microinjection zebrafish gentamicin acute kidney injury |
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