University of Pittsburgh, School of Medicine

4 ARTICLES PUBLISHED IN JoVE

Biology

Development of automated imaging and analysis for zebrafish chemical screens.

Andreas Vogt¹, Hiba Codore², Billy W. Day^3,4, Neil A. Hukriede², Michael Tsang²

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.

Biology

Intravenous Microinjections of Zebrafish Larvae to Study Acute Kidney Injury

Chiara Cianciolo Cosentino¹, Beth L. Roman², Iain A. Drummond³, Neil A. Hukriede¹

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.

Neuroscience

Modeling Fast-scan Cyclic Voltammetry Data from Electrically Stimulated Dopamine Neurotransmission Data Using QNsim1.0

Rashed Harun^1,2,3, Christine M. Grassi², 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.

Developmental Biology

A Simplified Method for Generating Kidney Organoids from Human Pluripotent Stem Cells

Aneta Przepiorski¹, Amanda E. Crunk¹, Teresa M. Holm², Veronika Sander², Alan J. Davidson², 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.