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Baylor College of Medicine (BCM)

16 ARTICLES PUBLISHED IN JoVE

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Biology

A Reversible, Non-invasive Method for Airway Resistance Measurements and Bronchoalveolar Lavage Fluid Sampling in Mice
Sumanth Polikepahad 1, Wade T. Barranco 1, Paul Porter 1, Bruce Anderson 2, Farrah Kheradmand 1,3, David B. Corry 1,3
1Department of Medicine, Baylor College of Medicine (BCM), 2Millenium Premier Group, 3Department of Immunology, Baylor College of Medicine (BCM)

Repeated measurements of rodent respiratory physiology and sampling of airway inflammatory cells are desirable, but generally not feasible. Here we describe a repeatable method for orally intubating mice that permits repeated measurements of airway hyperreactivity and sampling of airway inflammatory cells.

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Biology

Transverse Aortic Constriction in Mice
Angela C. deAlmeida 1, Ralph J. van Oort 1, Xander H.T. Wehrens 1,2
1Department of Molecular Physiology and Biophysics, Baylor College of Medicine (BCM), 2The Margaret M. and Albert B. Alkek Department of Medicine, Baylor College of Medicine (BCM)

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model to study mechanisms underlying cardiac hypertrophy and heart failure development. Here, we describe procedures to constrict the aorta to create a reproducible degree of cardiac hypertrophy in mice.

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Medicine

Programmed Electrical Stimulation in Mice
Na Li 1, Xander H.T Wehrens 1,2
1Department of Molecular Physiology and Biophysics, Baylor College of Medicine (BCM), 2The Margaret M. and Albert B. Alkek Department of Medicine, Baylor College of Medicine (BCM)

Programmed electrical stimulation provides the ability to determine conduction properties of the heart, and the possibility to induce and terminate cardiac arrhythmias using various pacing protocols. Using a transvenous catheter, intracardiac electrogram recordings can be obtained in mice following programmed electrical stimulation protocols to identify arrhythmogenic substrates.

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Medicine

Transthoracic Echocardiography in Mice
Jonathan L. Respress 1, Xander H.T. Wehrens 1,2
1Department of Molecular Physiology and Biophysics, Baylor College of Medicine (BCM), 2The Margaret M. and Albert B. Alkek Department of Medicine, Baylor College of Medicine (BCM)

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

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Biology

Ambulatory ECG Recording in Mice
Mark D. McCauley 1,2, Xander H.T Wehrens 1,2
1Department of Molecular Physiology and Biophysics, Baylor College of Medicine (BCM), 2The Margaret M. and Albert B. Alkek Department of Medicine, Baylor College of Medicine (BCM)

Telemetric ECG has emerged as an essential tool in evaluating animal models for cardiac arrhythmias and sudden cardiac death. Here, we present a stepwise guide to telemetric ECG recordings for application in long-term ambulatory ECG monitoring in mice.

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Biology

Imaging Cell Shape Change in Living Drosophila Embryos
Lauren Figard 1, Anna Marie Sokac 1,2
1Program in Cell & Molecular Biology, Baylor College of Medicine (BCM), 2Verna & Marrs McLean Department of Biochemistry & Molecular Biology, Baylor College of Medicine (BCM)

Early development of the fruit fly, Drosophila melanogaster, is characterized by a number of cell shape changes that are well suited for imaging approaches. This article will describe basic tools and methods required for live confocal imaging of Drosophila embryos, and will focus on a cell shape change called cellularization.

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Neuroscience

A Rapid Approach to High-Resolution Fluorescence Imaging in Semi-Thick Brain Slices
Jennifer Selever 1, Jian-Qiang Kong 2, Benjamin R. Arenkiel 3,4
1Department of Molecular & Human Genetics, Baylor College of Medicine (BCM), 2Precisionary Instruments Inc., 3Departments of Molecular & Human Genetics and Neuroscience, Baylor College of Medicine (BCM), 4Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital

Here we describe a rapid and simple method to image fluorescently labeled cells in semi-thick brain slices. By fixing, slicing, and optically clearing brain tissue we describe how standard epifluorescent or confocal imaging can be used to visualize individual cells and neuronal networks within intact nervous tissue.

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Neuroscience

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue
Kevin Ung 1, Benjamin R. Arenkiel 1,2,3
1Department of Molecular & Human Genetics, Baylor College of Medicine (BCM), 2Department of Neuroscience, Baylor College of Medicine (BCM), 3Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital

The development of optogenetics now provides the means to precisely stimulate genetically defined neurons and circuits, both in vitro and in vivo. Here we describe the assembly and implantation of a fiber optic for chronic photostimulation of brain tissue.

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Biology

Live Cell Imaging of Primary Rat Neonatal Cardiomyocytes Following Adenoviral and Lentiviral Transduction Using Confocal Spinning Disk Microscopy
Takashi Sakurai 1,2, Anthony Lanahan 2, Melissa J. Woolls 2, Na Li 2, Daniela Tirziu 2, Masahiro Murakami 2
1Max-Planck-Institute for Molecular Biomedicine and Institute of Cell Biology, 2Department of Internal Medicine, Yale Cardiovascular Research Center and Section of Cardiovascular Medicine

This protocol describes a method of live cell imaging using primary rat neonatal cardiomyocytes following lentiviral and adenoviral transduction using confocal spinning disk microscopy. This enables detailed observations of cellular processes in living cardiomyocytes.

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Neuroscience

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice
Gary Liu *1,2, Jay M. Patel *2,3, Burak Tepe 1, Cynthia K. McClard 2,4, Jessica Swanson 4, Kathleen B. Quast 4, Benjamin R. Arenkiel 1,3,4,5
1Program in Developmental Biology, Baylor College of Medicine, 2Medical Scientist Training Program, Baylor College of Medicine, 3Department of Neuroscience, Baylor College of Medicine, 4Department of Molecular and Human Genetics, Baylor College of Medicine, 5Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

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Biology

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay
Courtney M. Anderson 1, Annelies Laeremans 1, Xiao-Ming Mindy Wang 1, Xingyong Wu 1, Bingqing Zhang 1, Emerald Doolittle 1, Jeffrey Kim 1, Na Li 1, Helly Xiao Yan Pimentel 1, Emily Park 1, Xiao-Jun Ma 1
1Advanced Cell Diagnostics, Inc

Here, we describe an in situ hybridization assay which enables sensitive and specific detection of sequences as short as 50 nucleotides with single-nucleotide resolution at the single-cell level. The assay, which can be performed manually or automatically, can enable visualization of splice variants, short sequences, and mutations within the tissue context.

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Cancer Research

Preparation of Mitochondria from Ovarian Cancer Tissues and Control Ovarian Tissues for Quantitative Proteomics Analysis
Xianquan Zhan 1,2,3,4, Huanni Li 5, Shehua Qian 1,2,3, Xiaohan Zhan 1,2,3, Na Li 1,2,3
1Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, 2Hunan Engineering Laboratory for Structural Biology and Drug Design, Xiangya Hospital, Central South University, 3State Local Joint Engineering Laboratory for Anticancer Drugs, Xiangya Hospital, Central South University, 4National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, 5Department of Obstetrics and Gynecology, Xiangya Hospital, Central South University

This article presents a protocol of differential-speed centrifugation in combination with density gradient centrifugation to separate mitochondria from human ovarian cancer tissues and control ovarian tissues for quantitative proteomics analysis, resulting in a high-quality mitochondrial sample and high-throughput and high-reproducibility quantitative proteomics analysis of a human ovarian cancer mitochondrial proteome.

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Cancer Research

Analysis of Side Population in Solid Tumor Cell Lines
Xiaoli Dong 1, Yingying Wei 1, Tao Xu 1, Xiaoyue Tan 1,2,3, Na Li 1,2,3
1School of Medicine, Nankai University, 2Tianjin Key Laboratory of Tumour Microenvironment and Neurovascular Regulation, 3Collaborative Innovation Center for Biotherapy, Nankai University

A convenient, fast, and cost-effective method to measure the proportion of side population cells in solid tumor cell lines is presented.

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Developmental Biology

Imaging Intranuclear Actin Rods in Live Heat Stressed Drosophila Embryos
Natalie Biel 1,2, Lauren Figard 3, Anna Marie Sokac 1,2,3
1Integrative Molecular and Biomedical Sciences, Baylor College of Medicine, 2Department of Cell and Molecular Biology, School of Molecular and Cellular Biology, University of Illinois at Urbana-Champaign, 3Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine

The goal of this protocol is to inject Rhodamine-conjugated globular actin into Drosophila embryos and image intranuclear actin rod assembly following heat stress.

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Immunology and Infection

Cigarette Smoke Exposure in Mice using a Whole-Body Inhalation System
Daniel E. Morales-Mantilla 1,2, Xinyan Huang 3,7, Philip Erice 1,3, Paul Porter 4, Yun Zhang 1,5, Mary Figueroa 6, Joya Chandra 6, Katherine Y. King 2, Farrah Kheradmand 4,8, Antony Rodríguez 3,8
1Program in Immunology, Baylor College of Medicine, 2Department of Pediatrics, Section of Infectious Diseases, Baylor College of Medicine, 3Department of Medicine - Immunology Allergy and Rheumatology, Baylor College of Medicine, 4Department of Medicine, Pulmonary, Critical Care, Sleep Medicine, Baylor College of Medicine, 5Department of Pathology and Immunology, Baylor College of Medicine, 6Department of Pediatrics, Research and Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, 7Division of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Sun Yat-sen University, 8Center for Translational Research on Inflammatory Diseases (CTRID), Michael E. DeBakey VA Medical Center

This protocol demonstrates the study of the pathophysiologic effects of cigarette smoke (CS) with a whole-body inhalation (WBI) exposure system (WBIS) built in-house. This system can expose animals to CS under controlled repeatable conditions for research of CS-mediated effects on lung emphysema and hematopoiesis.

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Neuroscience

Imaging and Quantification of Intact Neuronal Dendrites via CLARITY Tissue Clearing
Brandon T. Pekarek 1, Patrick J. Hunt 1,2, Benjamin D. W. Belfort 1,2, Gary Liu 2, Benjamin R. Arenkiel 1,3
1Department of Genetics and Genomics, Baylor College of Medicine, 2Medical Scientist Training Program, Baylor College of Medicine, 3Department of Neuroscience, Baylor College of Medicine

Neuronal dendritic morphology often underlies function. Indeed, many disease processes that affect the development of neurons manifest with a morphological phenotype. This protocol describes a simple and powerful method for analyzing intact dendritic arbors and their associated spines.

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