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University of Colorado

18 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Assaying β-amyloid Toxicity using a Transgenic C. elegans Model
Vishantie Dostal 1, Christopher D. Link 1,2
1Institute for Behavioral Genetics, University of Colorado, 2Integrative Physiology, University of Colorado

The intensely studied nematode worm Caenorhabditis elegans can be transgenically engineered to express the human β-amyloid peptide (Aβ). Induced expression of Aβ in C. elegans muscle leads to a rapid, reproducible paralysis phenotype that can be used to monitor treatments that modulate Aβ toxicity.

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Medicine

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice
Michael Koeppen 1, Tobias Eckle 1, Holger K. Eltzschig 1
1Department of Anesthesiology, University of Colorado

A murine model for ventilator induced lung injury is an important tool to study an acute lung injury in vivo. Here, we report an easy applicable in situ model for acute lung injury using high-pressure mechanical ventilation to induce acute failure of the lung.

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Medicine

Use of a Hanging Weight System for Coronary Artery Occlusion in Mice
Tobias Eckle 1, Michael Koeppen 1, Holger Eltzschig 1
1Department of Anesthesiology, University of Colorado Denver

A murine model for myocardial ischemia and ischemic preconditioning is an important tool study cardioprotective mechanisms in vivo. Here, we report an easy applicable in situ model for cardiac IP using a hanging-weight system for coronary artery occlusion.

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Medicine

Use of a Hanging-weight System for Isolated Renal Artery Occlusion
Almut Grenz 1, Julee H. Hong 1, Alexander Badulak 1, Douglas Ridyard 1, Timothy Luebbert 2, Jae-Hwan Kim 3, Holger K. Eltzschig 1
1Department of Anesthesiology, University of Colorado, 2School of Medicine, University of Colorado, 3Department of Anesthesiology, Korea University College of Medicine

A precise murine model for acute kidney injury (AKI) due to ischemia is an important tool to investigate acute kidney injury and possibly find therapeutic tools to treat renal injury. The hanging weight system offers a tool for immediate and reliable renal artery occlusion and reperfusion without causing renal congestion.

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Medicine

Use of a Hanging-weight System for Liver Ischemia in Mice
Michael Zimmerman 1, Eunyoung Tak 2, Maria Kaplan 1, Mercedes Susan Mandell 2, Holger K. Eltzschig 2, Almut Grenz 2
1UCH Transplant Center, University of Colorado, Denver, 2Department of Anesthesiology, University of Colorado, Denver

We established a novel murine model of a hanging weight system for portal triad occlusion. This technique may be useful for future investigations of ischemia in murine hepatic models.

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Medicine

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion
Marshall T. Bell 1, T. Brett Reece 1, Phillip D. Smith 1, Joshua Mares 1, Michael J. Weyant 1, Joseph C. Cleveland Jr. 1, Kirsten A. Freeman 1, David A. Fullerton 1, Ferenc Puskas 2
1Department of Surgery, Division of Cardiothoracic Surgery, University of Colorado, 2Department of Anesthesiology, University of Colorado

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

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Environment

High-throughput Fluorometric Measurement of Potential Soil Extracellular Enzyme Activities
Colin W. Bell 1, Barbara E. Fricks 1, Jennifer D. Rocca 1, Jessica M. Steinweg 2, Shawna K. McMahon 3, Matthew D. Wallenstein 1
1Natural Resource Ecology Laboratory, Colorado State University, 2Biosciences Division, Oak Ridge National Laboratory, 3Department of Bioengineering, University of Colorado

To measure potential rates of soil extracellular enzyme activities, synthetic substrates that are bound to a fluorescent dye are added to soil samples. Enzyme activity is measured as the fluorescent dye is released from the substrate by an enzyme-catalyzed reaction, where higher fluorescence indicates more substrate degradation.

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Bioengineering

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing
Shama Ahmad 1, Aftab Ahmad 1, Keith B. Neeves 2, Tara Hendry-Hofer 1, Joan E. Loader 1, Carl W. White 1, Livia Veress 1
1Pediatric Airway Research Center, Department of Pediatrics, University of Colorado, 2Department of Chemical and Biological Engineering, Colorado School of Mines

This protocol is designed to demonstrate exposure method of cell cultures to inhaled toxic chemicals. Exposure of differentiated air-liquid interface (ALI) cultures of airway epithelial cells provides a unique model of airway exposure to toxic gases such as chlorine. In this manuscript we describe effect of chlorine exposure on air-liquid interface cultures of epithelial cells and submerged culture of cardiomyocytes. In vitro exposure systems allow important mechanistic studies to evaluate pathways that could then be utilized to develop novel therapeutic agents.

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Biology

Purifying the Impure: Sequencing Metagenomes and Metatranscriptomes from Complex Animal-associated Samples
Yan Wei Lim 1, Matthew Haynes 2, Mike Furlan 1, Charles E. Robertson 3, J. Kirk Harris 4, Forest Rohwer 1
1Department of Biology, San Diego State University, 2DOE Joint Genome Institute, 3Department of Molecular, Cellular and Developmental Biology, University of Colorado, 4Department of Pediatrics, School of Medicine, University of Colorado

Using the cystic fibrosis airway as an example, the manuscript presents a comprehensive workflow comprising a combination of metagenomic and metatranscriptomic approaches to characterize the microbial and viral communities in animal-associated samples.

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Neuroscience

Receptor Autoradiography Protocol for the Localized Visualization of Angiotensin II Receptors
Andrea Linares 1, Leena E. Couling 2, Eduardo J. Carrera 3, Robert C. Speth 2
1Farquhar College of Arts and Sciences, Nova Southeastern University, 2Department of Pharmaceutical Sciences, College of Pharmacy, Nova Southeastern University, 3School of Medicine, University of Colorado

Here we present a protocol to describe the localization of angiotensin II Type 1 receptors in the rat brain by quantitative, densitometric, in vitro receptor autoradiography using an iodine-125 labeled analog of angiotensin II.

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Neuroscience

Spinal Cord Neurons Isolation and Culture from Neonatal Mice
Mohamed Eldeiry 1, Katsuhiro Yamanaka 1, T. Brett Reece 1, Muhammad Aftab 1
1Department of Surgery, Division of Cardiothoracic Surgery, University of Colorado Anschutz Medical Campus

This study presents a technique for the isolation of neurons from WT neonatal mice. It requires the careful dissection of the spinal cord from the neonatal mouse, followed by the separation of neurons from the spinal cord tissue through mechanical and enzymatic cleavage.

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Environment

Experimental Methods of Dust Charging and Mobilization on Surfaces with Exposure to Ultraviolet Radiation or Plasmas
Xu Wang 1,2, Joseph Schwan 1,2, Noah Hood 1,2, Hsiang-Wen Hsu 1,2, Eberhard Grün 1,2, Mihály Horányi 1,2
1Laboratory for Atmospheric and Space Physics, University of Colorado, 2NASA/SSERVI's Institute for Modeling Plasma, Atmospheres and Cosmic Dust

Dust charging and mobilization is demonstrated in three experiments with exposure to thermal plasma with beam electrons, beam electrons only, or ultraviolet (UV) radiation only. These experiments present the advanced understanding of electrostatic dust transport and its role in shaping the surfaces of airless planetary bodies.

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Neuroscience

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method
Weiyue Zhang *1, Wei Zhang *1, Ning Dai *1, Chenxia Han 2, Fengzhi Wu 1, Xu Wang 1, Libo Tan 1, Jie Li 1, Feng Li 1, Qingjia Ren 3
1School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, 2Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, 3Institute of Tibetan Medicine, Tibetan Traditional Medical College

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

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Medicine

Generation of Human 3D Lung Tissue Cultures (3D-LTCs) for Disease Modeling
Michael Gerckens 1,2,3, Hani N. Alsafadi 4,5,6, Darcy E. Wagner 4,5,6, Michael Lindner 2,7, Gerald Burgstaller *1,2,3, Melanie Königshoff *1,2,3,8
1Comprehensive Pneumology Center, Ludwig-Maximilians-Universität and Helmholtz Zentrum Munich, 2German Center of Lung Research (DZL), 3Translational Lung Research and CPC-M bioArchive, Helmholtz Zentrum München, Comprehensive Pneumology Center Munich DZL/CPC-M, 4Department of Experimental Medical Science, Lung Bioengineering and Regeneration, Lund University, 5Wallenberg Center for Molecular Medicine, Lund University, 6Stem Cell Centre, Lund University, 7Asklepios Fachkliniken Munich-Gauting, 8Department of Medicine, Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado

Here, we present a protocol for the preparation of agarose-filled human precision-cut lung slices from resected patient tissue that are suitable for generating 3D lung tissue cultures to model human lung diseases in biological and biomedical studies.

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Cancer Research

Patient-derived Heterogeneous Xenograft Model of Pancreatic Cancer Using Zebrafish Larvae as Hosts for Comparative Drug Assessment
Lei Wang *1, Huan Chen *2,3, Fei Fei *1, Xianfeng He 2,3, Shaoyang Sun 1, Kunpeng Lv 1, Bo Yu 2,3, Jiang Long 2,4,5,6, Xu Wang 1
1Department of Biochemistry and Molecular Biology, Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, School of Basic Medical Sciences, Fudan University, 2National Human Genetic Resources Sharing Service Platform (2005DKA21300), 3Fudan University Shanghai Cancer Center, 4Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, 5Department of Oncology, Shanghai Medical College, Fudan University, 6Pancreatic Cancer Institute, Fudan University

This protocol describes optimization procedures in a virus-based dual fluorescence-labeled tumor xenograft model using larval zebrafish as hosts. This heterogeneous xenograft model mimics the tissue composition of pancreatic cancer microenvironment in vivo and serves as a more precise tool for assessing drug responses in personalized zPDX (zebrafish patient-derived xenograft) models.

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Medicine

Ultrasonography of the Adult Male Urinary Tract for Urinary Functional Testing
Teresa T. Liu *1,2, Allison C. Rodgers *3, Tristan M. Nicholson 4, Jill A. Macoska 5,6, Paul C. Marker 6,7, Chad M. Vezina 6,8, Dale E. Bjorling 6,8, Alejandro Roldan-Alzate 2,9,10, Diego Hernando 10,11, Granville L. Lloyd 12, Timothy A. Hacker 3, William A. Ricke 1,6
1Department of Urology, University of Wisconsin-Madison, 2K12 Kure, University of Wisconsin-Madison, 3Cardiovascular Research Center, Department of Medicine, University of Wisconsin-Madison, 4Department of Urology, University of Washington, 5University of Massachusetts Boston, 6U54 George M. O'Brien Center, University of Wisconsin-Madison, 7College of Pharmacy, University of Wisconsin-Madison, 8School of Veterinary Medicine, University of Wisconsin-Madison, 9Department of Mechanical Engineering, University of Wisconsin-Madison, 10Department of Radiology, University of Wisconsin-Madison, 11Department of Medical Physics, University of Wisconsin-Madison, 12Rocky Mountain Regional VA Medical Center, University of Colorado

We describe the use of high frequency ultrasound with contrast imaging as a method to measure bladder volume, bladder wall thickness, urine velocity, void volume, void duration, and urethral diameter. This strategy can be used to assess voiding dysfunction and treatment efficacy in various mouse models of lower urinary tract dysfunction (LUTD).

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Biochemistry

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling
Steven E. Guard 1, Christopher C. Ebmeier 1, William M. Old 1,2
1Department of Molecular, Cellular and Developmental Biology, University of Colorado, 2Linda Crnic Institute for Down Syndrome, University of Colorado School of Medicine

Described is a proteomics workflow for identifying protein interaction partners from a nuclear subcellular fraction using immunoaffinity enrichment of a given protein of interest and label-free mass spectrometry. The workflow includes subcellular fractionation, immunoprecipitation, filter aided sample preparation, offline cleanup, mass spectrometry, and a downstream bioinformatics pipeline.

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Neuroscience

Miniscope Recording Calcium Signals at Hippocampus of Mice Navigating an Odor Plume
Fabio M. Simoes de Souza 1, Ryan Williamson 2, Connor McCullough 3, Alec Teel 1, Gregory Futia 3, Ming Ma 1, Aaron True 4, John P. Crimaldi 4, Emily Gibson 3,5, Diego Restrepo 1,5
1Department of Cell and Developmental Biology, University of Colorado Anschutz Medical Campus, 2Neurotechnology Center, University of Colorado Anschutz Medical Campus, 3Department of Bioengineering, University of Colorado Anschutz Medical Campus, 4Civil, Environmental & Architectural Engineering, University of Colorado, 5Neuroscience Graduate Program, University of Colorado Anschutz Medical Campus

This protocol investigates the brain-behavior relationship in hippocampal CA1 in mice navigating an odor plume. We provide a step-by-step protocol, including surgery to access imaging of the hippocampus, behavioral training, miniscope GCaMP6f recording and processing of the brain, and behavioral data to decode the mouse position from ROI neural activity.

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