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University of Pittsburgh, School of Medicine

4 ARTICLES PUBLISHED IN JoVE

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Biology

Development of automated imaging and analysis for zebrafish chemical screens.
Andreas Vogt 1, Hiba Codore 2, Billy W. Day 3,4, Neil A. Hukriede 2, Michael Tsang 2
1Pharmacology and Chemical Biology, University of Pittsburgh Drug Discovery Institute, 2Department of Microbiology and Molecular Genetics, University of Pittsburgh, 3Department of Pharmaceutical Sciences, University of Pittsburgh, 4Department of Chemistry, University of Pittsburgh

We report the development of a system for automated imaging and analysis of zebrafish transgenic embryos in multiwell plates. This demonstrates the ability to measure dose dependent effects of a small molecule, BCI, on Fibroblast Growth Factor reporter gene expression and provide technology for establishing high-throughput zebrafish chemical screens.

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Biology

Intravenous Microinjections of Zebrafish Larvae to Study Acute Kidney Injury
Chiara Cianciolo Cosentino 1, Beth L. Roman 2, Iain A. Drummond 3, Neil A. Hukriede 1
1Department of Developmental Biology, University of Pittsburgh, 2Department of Biological Sciences, University of Pittsburgh, 3Department of Medicine and Genetics, Harvard Medical School

We describe a technique of microinjecting the aminoglycoside, gentamicin, into 2 days post-fetilization (dpf) zebrafish larvae to induce acute kidney injury (AKI). We also describe a method for whole mount immunohistochemistry, plastic embedding and sectioning of zebrafish larvae to visualize the AKI mediated damage.

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Neuroscience

Modeling Fast-scan Cyclic Voltammetry Data from Electrically Stimulated Dopamine Neurotransmission Data Using QNsim1.0
Rashed Harun 1,2,3, Christine M. Grassi 2, Miranda J. Munoz 2,4, Amy K. Wagner 1,2,3
1Center for Neuroscience, University of Pittsburgh, 2Department of Physical Medicine & Rehabilitation, University of Pittsburgh, School of Medicine, 3Safar Center for Resuscitation Research, University of Pittsburgh, 4Department of Biological Sciences, Mellon College of Science, Carnegie Mellon University

Fast-scan cyclic voltammetry can monitor in vivo dopamine neurotransmission in the context of drugs, disease, and other experimental manipulations. This work describes the implementation of QNsim1.0, a software to model electrically stimulated dopamine responses according to the quantitative neurobiological model to quantify estimates of dopamine release and reuptake dynamics.

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Developmental Biology

A Simplified Method for Generating Kidney Organoids from Human Pluripotent Stem Cells
Aneta Przepiorski 1, Amanda E. Crunk 1, Teresa M. Holm 2, Veronika Sander 2, Alan J. Davidson 2, Neil A. Hukriede 1,3
1Department of Developmental Biology, University of Pittsburgh, School of Medicine, 2Department of Molecular Medicine and Pathology, School of Medical Sciences, University of Auckland, 3Center for Critical Care Nephrology, University of Pittsburgh, School of Medicine

Here we describe a protocol to generate kidney organoids from human pluripotent stem cells (hPSCs). This protocol generates kidney organoids within two weeks. The resulting kidney organoids can be cultured in large-scale spinner flasks or multi-well magnetic stir plates for parallel drug-testing approaches.

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