S'identifier

Emory University

36 ARTICLES PUBLISHED IN JoVE

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Biology

Obtaining Eggs from Xenopus laevis Females
Marie K. Cross 1, Maureen Powers 1
1Department of Cell Biology, Emory University

The eggs of Xenopus laevis intact, lysed, and/or fractionated are useful for a wide variety of experiments. This protocol shows how to induce egg laying, collect and dejelly the eggs, and sort the eggs to remove any damaged eggs.

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Biology

Preparation and Fractionation of Xenopus laevis Egg Extracts
Marie K. Cross 1, Maureen Powers 1
1Department of Cell Biology, Emory University

Crude and fractionated Xenopus egg extracts can be used to provide ingredients for reconstituting cellular processes for morphological and biochemical analysis. Egg lysis and differential centrifugation are used to prepare the crude extract which in turn in used to prepare fractionated extracts and light membrane preparations.

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Biology

In Vitro Nuclear Assembly Using Fractionated Xenopus Egg Extracts
Marie Cross 1, Maureen Powers 1
1Department of Cell Biology, Emory University

Nuclear membrane assembly is an essential step in the cell division cycle; this process can be replicated in the test tube by combining Xenopus sperm chromatin, cytosol, and light membrane fractions. Complete nuclei are formed, including nuclear membranes with pore complexes, and these reconstituted nuclei are capable of normal nuclear processes.

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Biology

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
Stephanie A. Zlatic 1, Pearl V. Ryder 1, Gloria Salazar 2, Victor Faundez 1
1Department of Cell Biology, Emory University, 2Department of Medicine, Division of Cardiology, Emory University

The cell permeable crosslinker DSP [dithiobis-(succinimidyl propionate)] stabilizes transient and labile interactions in vivo, which allows their isolation using stringent protein complex purification techniques. Here we present a technique for crosslinking cells grown in culture followed by isolation of protein complexes by immunoprecipitation.

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Medicine

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium
Douglas Nam 1, Chih-Wen Ni 2, Amir Rezvan 1, Jin Suo 2, Klaudia Budzyn 1, Alexander Llanos 1, David G. Harrison 1, Don P. Giddens 2, Hanjoong Jo 1,2,3
1Department of Medicine, Division of Cardiology, Emory University, 2Coulter Department of Biomedical Engineering, Georgia Tech and Emory University, 3Department of Bioinspired Science, Ewha Womans University

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

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Neuroscience

Simultaneous fMRI and Electrophysiology in the Rodent Brain
Wen-ju Pan 1,2, Garth Thompson 1,2, Matthew Magnuson 1,2, Waqas Majeed 1,2, Dieter Jaeger 3, Shella Keilholz 1,2
1Biomedical Engineering, Emory University, 2Biomedical Engineering, Georgia Institute of Technology, 3Biology, Emory University

We have developed a method for simultaneous functional magnetic resonance imaging and electrophysiological recording in the rodent brain, providing a platform for the investigation of the relationship between neural activity and the blood oxygenation level dependent (BOLD) MRI signal.

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Neuroscience

SDS-PAGE/Immunoblot Detection of Aβ Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval
Rebecca F. Rosen 1, Yasushi Tomidokoro 2, Jorge A. Ghiso 3, Lary C. Walker 1,4
1Yerkes National Primate Research Center, Emory University, 2Department of Neurology, Institute of Clinical Medicine, Tsukuba University, 3Department of Pathology, New York University School of Medicine, 4Department of Neurology, Emory University

We describe a technique for the preparation of clarified human cortical homogenates, protein separation by SDS-PAGE, antigen retrieval and immunoblotting with an antibody to the Aβ peptide. Using this protocol, we consistently detect monomeric and multimeric Aβ in cortical tissue from humans with Alzheimer's pathology.

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Biology

Direct Tracheal Instillation of Solutes into Mouse Lung
My N. Helms 1,2, Edilson Torres-Gonzalez 2,3, Preston Goodson 1, Mauricio Rojas 2,3
1Department of Physiology, Emory University, 2Center for Respiratory Health, Emory University, 3Department of Medicine, Emory University

Intratracheal instillations deliver solutes directly into the lungs. This procedure targets the delivery of the instillate into the distal regions of the lung, and is therefore often incorporated in studies aimed at studying alveoli. We provide a detailed survival protocol for performing intratracheal instillations in mice.

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Biology

Competitive Genomic Screens of Barcoded Yeast Libraries
Andrew M. Smith *1,2, Tanja Durbic *2,3, Julia Oh *4, Malene Urbanus 1,2, Michael Proctor 5, Lawrence E. Heisler 2,3, Guri Giaever 2,6, Corey Nislow 1,2,3
1Banting and Best Department of Medical Research and Department of Molecular Genetics, University of Toronto, 2Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 3Donnelly Sequencing Centre, University of Toronto, 4Genetics and Molecular Biology Branch, National Human Genome Research Institute, NIH, 5Stanford Genome Technology Center, Stanford School of Medicine, Stanford University , 6Department of Pharmaceutical Sciences, University of Toronto

We have developed comprehensive, unbiased genome-wide screens to understand gene-drug and gene-environment interactions. Methods for screening these mutant collections are presented.

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Immunology and Infection

Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine
Duke Geem 1, Oscar Medina-Contreras 1, Wooki Kim 2, Clifton S. Huang 1, Timothy L. Denning 1,2
1Department of Pediatrics, Emory University, 2Department of Pathology & Laboratory Medicine, Emory University

Here, we detail a methodology for the rapid isolation of mouse intestinal dendritic cells (DCs) and macrophages. Phenotypic characterization of intestinal DCs and macrophages is performed using multi-color flow cytometric analysis while magnetic bead enrichment followed by cell sorting is used to yield highly pure populations for functional studies.

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Engineering

Compact Quantum Dots for Single-molecule Imaging
Andrew M. Smith 1, Shuming Nie 1,2
1Department of Biomedical Engineering, Emory University, 2Department of Chemistry, Georgia Institute of Technology

We describe the preparation of colloidal quantum dots with minimized hydrodynamic size for single-molecule fluorescence imaging. Compared to conventional quantum dots, these nanoparticles are similar in size to globular proteins and are optimized for single-molecule brightness, stability against photodegradation, and resistance to nonspecific binding to proteins and cells.

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Medicine

Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs
Thais Federici 1, Carl V. Hurtig 1, Kentrell L. Burks 1, Jonathan P. Riley 1, Vibhor Krishna 2, Brandon A. Miller 1, Eric A. Sribnick 1, Joseph H. Miller 3, Natalia Grin 1, Jason J. Lamanna 1,4,5, Nicholas M. Boulis 1
1Department of Neurosurgery, Emory University, 2Department of Neuroscience, Medical University of South Carolina, 3Division of Neurosurgery, University of Alabama, Birmingham, 4Department of Biomedical Engineering, Georgia Institute of Technology , 5Department of Biomedical Engineering, Emory University

Short visual description of the surgical technique and device used for the delivery of (gene and cell) therapies into the spinal cord. The technique is demonstrated in the animal but is entirely translatable and currently being used for human application.

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Neuroscience

A Lightweight, Headphones-based System for Manipulating Auditory Feedback in Songbirds
Lukas A. Hoffmann 1,2, Conor W. Kelly 1,3, David A. Nicholson 1,2, Samuel J. Sober 1
1Department of Biology, Emory University, 2Neuroscience Graduate Program, Emory University, 3Program in Neuroscience and Behavioral Biology, Emory University

We describe the design and assembly of miniaturized headphones suitable for replacing a songbird’s natural auditory feedback with a manipulated acoustic signal. Online sound processing hardware is used to manipulate song output, introduce real-time errors in auditory feedback via the headphones, and drive vocal motor learning.

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Immunology and Infection

A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
Daniel T. Claiborne *1, Jessica L. Prince *1, Eric Hunter 1,2
1Emory Vaccine Center at Yerkes National Primate Research Center, Emory University, 2Department of Pathology and Laboratory Medicine, Emory University

HIV-1 pathogenesis is defined by both viral characteristics and host genetic factors. Here we describe a robust method that allows for reproducible measurements to assess the impact of the gag gene sequence variation on the in vitro replication capacity of the virus.

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Immunology and Infection

Pseudomonas aeruginosa Induced Lung Injury Model
Varsha Suresh Kumar 1, Ruxana T. Sadikot 2, Jeanette E. Purcell 3, Asrar B. Malik 1, Yuru Liu 1
1Department of Pharmacology, University of Illinois College of Medicine, University of Illinois at Chicago, 2Section of Pulmonary and Critical Care Medicine, Atlanta VAMC, Emory University, 3Biologic Resources Lab, University of Illinois at Chicago

We have developed a mouse lung injury model by intra-tracheal injection of bacteria Pseudomonas aeruginosa. This model mimics lung injury during pneumonia and is clinically relevant.

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Medicine

Real-time X-ray Imaging of Lung Fluid Volumes in Neonatal Mouse Lung
Ashley E. Van Avermaete 1, Phi T. Trac 1, Theresa W. Gauthier 1, My N. Helms 2
1Department of Pediatrics, Neonatology Division, Emory Children's Center, Emory University, 2Department of Internal Medicine, Pulmonary Division, University of Utah

We present a protocol to assess the rate of alveolar fluid clearance or pulmonary edema in neonatal mouse lung using X-ray imaging technology.

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Environment

Measuring Rates of Herbicide Metabolism in Dicot Weeds with an Excised Leaf Assay
Rong Ma 1, Joshua J. Skelton 1, Dean E. Riechers 1
1Department of Crop Sciences, University of Illinois

This manuscript describes how herbicide metabolism rates can be effectively quantified with excised leaves from a dicot weed, thereby reducing variability and removing any possible confounding effects of herbicide uptake or translocation typically observed in whole-plant assays.

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Chemistry

Extraction and Purification of Polyphenols from Freeze-dried Berry Powder for the Treatment of Vascular Smooth Muscle Cells In Vitro
Rafaela G. Feresin 1, Shirin Pourafshar 2,3, Jingwen Huang 2, Yitong Zhao 2, Bahram H. Arjmandi 2,3, Gloria Salazar 2,3
1Department of Dietetics and Nutrition, University of Arkansas for Medical Sciences, 2Department of Nutrition, Food and Exercise Sciences, Florida State University, 3Center for Advancing Exercise and Nutrition Research on Aging (CAENRA), Florida State University

This work details a step-by-step method to prepare polyphenol-rich extracts from freeze-dried berry powder. In addition, it provides a thorough description of how to use these polyphenol-rich extracts in cell culture in the presence of the peptide hormone angiotensin II (Ang II) using Vascular Smooth Muscle Cells (VSMCs).

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Biochemistry

Analyses of Mitochondrial Calcium Influx in Isolated Mitochondria and Cultured Cells
Joshua T. Maxwell 1, Chin-Hsien Tsai 1, Tahmina A. Mohiuddin 2, Jennifer Q. Kwong 1
1Department of Pediatrics, Emory University School of Medicine, 2Emory College of Arts and Sciences, Emory University

Here, we present two protocols for the measurement of mitochondrial Ca2+ influx in isolated mitochondria and cultured cells. For isolated mitochondria, we detail a plate reader-based Ca2+ import assay using the Ca2+ sensitive dye calcium green-5N. For cultured cells, we describe a confocal microscopy method using the Ca2+ dye Rhod-2/AM.

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Genetics

Bronchoalveolar Lavage Exosomes in Lipopolysaccharide-induced Septic Lung Injury
Zhihong Yuan 1,2, Brahmchetna Bedi 1, Ruxana T. Sadikot 1,2
1Department of Veterans Affairs, Atlanta VAMC, 2Division of Pulmonary and Critical Care Medicine, Department of Medicine, Emory University

Mice exposed to intraperitoneal LPS secrete exosomes in Broncho-alveolar lavage (BAL) fluid that are packaged with miRNAs. Using a co-culture system, we show that exosomes released in the BAL fluid disrupt expression of tight junction proteins in bronchial epithelial cells and increase expression of pro-inflammatory cytokines that accentuate lung injury.

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Chemistry

Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery
Yang Wu *1, Wei Zhang *2, Jian Zhang 2, Zhi-Xiang Mao 3, Li Ding 2, Hao Li 3, Rong Ma 1, Jin-Hai Tang 2
1Research Center of Clinical Oncology, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & Nanjing Medical University Affiliated Cancer Hospital, 2Department of General Surgery, The First Affiliated Hospital with Nanjing Medical University, 3School of Clinical Medicine, Xuzhou Medical University

This work presents the preparation of methionine functionalized biocompatible block copolymers (mBG) via the reversible addition-fragmentation chain transfer (RAFT) method. The plasmid DNA complexing ability of the obtained mBG and their transfection efficiency were also investigated. The RAFT method is very beneficial for polymerizing monomers containing special functional groups.

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Neuroscience

Culture of Neurospheres Derived from the Neurogenic Niches in Adult Prairie Voles
Daniela Ávila-González 1, Larry J. Young 2, Francisco Camacho 1, Raúl G. Paredes 1,3, Néstor F. Díaz 4, Wendy Portillo 1
1Departamento de Neurobiología Conductual y Cognitiva, Instituto de Neurobiología, Universidad Nacional Autónoma de México, 2Silvio O. Conte Center for Oxytocin and Social Cognition, Center for Translational Social Neuroscience, Department of Psychiatry and Behavioral Sciences, Yerkes National Primate Research Center, Emory University, 3Escuela Nacional de Estudios Superiores Juriquilla, Universidad Nacional Autónoma de México, 4Departamento de Fisiología y Desarrollo Celular, Instituto Nacional de Perinatología

We established the conditions to culture neural progenitor cells from the subventricular zone and dentate gyrus of the adult brain of prairie voles, as a complementary in vitro study, to analyze the sex-dependent differences between neurogenic niches that could be part of functional plastic changes associated with social behaviors.

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Neuroscience

Flow Cytometry Approach to Characterize Phagocytic Properties of Acutely-Isolated Adult Microglia and Brain Macrophages In Vitro
Supriya Ramesha 1, Sruti Rayaprolu 1, Srikant Rangaraju 1
1Department of Neurology, Emory University

Assessment of phagocytic properties of microglia and brain macrophages can provide a valuable functional dimension to molecular profiling studies. We describe a validated protocol that uses flow cytometry to rapidly and reliably quantify phagocytosis of fluorescent microspheres and amyloid beta fibrils by acutely-isolated microglia and brain macrophages from mouse models.

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Biology

Contribution of the Na+/K+ Pump to Rhythmic Bursting, Explored with Modeling and Dynamic Clamp Analyses
Ricardo Javier Erazo-Toscano 1,2, Parker J. Ellingson 2, Ronald L. Calabrese *1, Gennady S. Cymbalyuk *2
1Department of Biology, Emory University, 2Neuroscience Institute, Georgia State University

Presented here is a method for investigation of the roles of the Na+/K+ pump and persistent Na+ current in leech heart interneurons using dynamic clamp.

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Bioengineering

In vivo Structural Assessments of Ocular Disease in Rodent Models using Optical Coherence Tomography
Rachael S. Allen 1,2, Katie Bales 1,3, Andrew Feola 1,2, Machelle T Pardue 1,2,3
1Center of Excellence for Visual and Neurocognitive Rehabilitation, Atlanta Veterans Affairs Medical Center, 2Department of Biomedical Engineering, Georgia Institute of Technology, 3Department of Ophthalmology, Emory University

Here, we describe the use of spectral-domain optical coherence tomography (SD-OCT) to visualize retinal and ocular structures in vivo in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia.

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Behavior

Behavioral Assessment of Visual Function via Optomotor Response and Cognitive Function via Y-Maze in Diabetic Rats
Kaavya Gudapati *1,2, Anayesha Singh *1,3, Danielle Clarkson-Townsend 1,4, Stephen Q. Phillips 1, Amber Douglass 1, Andrew J. Feola 1,2, Rachael S. Allen 1,2
1Center for Visual and Neurocognitive Rehabilitation, Atlanta VA Medical Center, 2Department of Biomedical Engineering, Georgia Institute of Technology, 3Department of Neuroscience, Emory University, 4Gangarosa Department of Environmental Health, Emory University

Neural degeneration in both eyes and brain as a result of diabetes can be observed through behavioral tests carried out on rodents. The Y-maze, a measure of spatial cognition, and the optomotor response, a measure of visual function, both provide insight into potential diagnoses and treatments.

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Genetics

Two Infection Assays to Study Non-Lethal Virulence Phenotypes in C. Albicans using C. Elegans
Amanda C. Smith 1, Judy Dinh 2, Meleah A. Hickman 1,2
1Graduate Program in Genetics & Molecular Biology, Emory University, 2Department of Biology, Emory University

Fungal opportunist pathogens can cause life-threatening as well as minor infections, but non-lethal phenotypes are frequently ignored when studying virulence. Therefore, we developed a nematode model that monitors both the survival and reproduction aspects of host to investigate fungal virulence.

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Bioengineering

DNA Tension Probes to Map the Transient Piconewton Receptor Forces by Immune Cells
Rong Ma 1, Anna V. Kellner 2, Yuesong Hu 1, Brendan R. Deal 1, Aaron T. Blanchard 2, Khalid Salaita 1,2
1Department of Chemistry, Emory University, 2Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University

This paper describes a detailed protocol for using DNA-based tension probes to image the receptor forces applied by immune cells. This approach can map receptor forces >4.7pN in real-time and can integrate forces over time.

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Biology

Isolation of Endothelial Cells from the Lumen of Mouse Carotid Arteries for Single-Cell Multi-Omics Experiments
Sandeep Kumar *1, Aitor Andueza *1, Nicolas Villa-Roel 1, Juyoung Kim 1, Dong-Won Kang 1, Hanjoong Jo 1,2
1Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 2Division of Cardiology, Georgia Institute of Technology and Emory University

We present a method for isolating endothelial cells and nuclei from the lumen of mouse carotid arteries exposed to stable or disturbed flow conditions to perform single-cell omics experiments.

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Neuroscience

Modeling Neonatal Intraventricular Hemorrhage Through Intraventricular Injection of Hemoglobin
Brandon A. Miller 1,2, Shelei Pan 3, Peter H. Yang 3, Catherine Wang 1, Amanda L. Trout 1, Dakota DeFreitas 3, Sruthi Ramagiri 3, Scott D. Olson 2, Jennifer M. Strahle 3,4,5
1Department of Neurosurgery, University of Kentucky, 2Department of Pediatric Surgery, University of Texas, 3Department of Neurological Surgery, Washington University in St. Louis School of Medicine, 4Department of Orthopedic Surgery, Washington University in St. Louis School of Medicine, 5Department of Pediatrics, Washington University in St. Louis School of Medicine

We present a model of neonatal intraventricular hemorrhage using rat pups that mimics the pathology seen in humans.

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Biology

Using Single-Worm Data to Quantify Heterogeneity in Caenorhabditis elegans-Bacterial Interactions
Megan N. Taylor 1, Satya Spandana Boddu 2, Nic M. Vega 1,2
1Department of Biology, Emory University, 2Department of Physics, Emory University

This protocol describes a 96-well disruption of individual bacterially colonized Caenorhabditis elegans following cold paralysis and surface bleaching to remove external bacteria. The resulting suspension is plated on agar plates to allow accurate, medium-throughput quantification of bacterial load in large numbers of individual worms.

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Environment

Visualizing Field Data Collection Procedures of Exposure and Biomarker Assessments for the Household Air Pollution Intervention Network Trial in India
Karthikeyan D. Rajamani 1, Sankar Sambandam 1, Krishnendu Mukhopadhyay 1, Naveen Puttaswamy 1, Gurusamy Thangavel 1, Durairaj Natesan 1, Rengaraj Ramasamy 1, Saritha Sendhil 1, Amudha Natarajan 1, Vigneswari Aravindalochan 1, Ajay Pillarisetti 2, Michael Johnson 3, Joshua Rosenthal *4, Kyle Steenland 5, Ricardo Piedhrahita 3, Jennifer Peel 6, Maggie L. Clark 6, Dana Boyd Barr 5, Sarah Rajkumar 6, Bonnie Young 6, Shirin Jabbarzadeh 7, Ghislaine Rosa 8, Miles Kirby 9, Lindsay J. Underhill 10, Anaite Diaz-Artiga 11, Amy Lovvorn 5, William Checkley 12, Thomas Clasen 5, Kalpana Balakrishnan 1
1Department of Environmental Health Engineering, ICMR Center for Advanced Research on Air Quality, Climate and Health, Faculty of Public Health, Sri Ramachandra Institute of Higher Education and Research (Deemed University), 2Division of Environmental Health Sciences, University of California, Berkeley, 3Berkeley Air Monitoring Group, 4Division of International Epidemiology and Population Studies, National Institutes of Health, 5Gangarosa Department of Environmental Health, Rollins School of Public Health, Emory University, 6Department of Environmental and Radiological Health Sciences, Colorado State University, 7Department of Biostatistics and Informatics, Rollins School of Public Health, Emory University, 8Department of Disease Control, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, 9Department of Global Health & Population, Harvard, T.H. Chan School of Public Health, 10Cardiovascular Division, Washington University School of Medicine, Washington University, 11Centro de Estudios en Salud, Universidad del Valle de Guatemala, 12Division of Pulmonary and Critical Care, School of Medicine, Johns Hopkins University

We detail the consistent, high-quality procedures used throughout air and biological sampling processes at Indian field sites during a large randomized controlled trial. Insights gathered from the oversight of applications of innovative technologies, adapted for exposure assessment in rural regions, enable better field data collection practices with more reliable outcomes.

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Neuroscience

Through a Dog's Eyes: fMRI Decoding of Naturalistic Videos from the Dog Cortex
Erin M. Phillips 1, Kirsten D. Gillette 1, Daniel D. Dilks 1, Gregory S. Berns 1
1Psychology Department, Emory University

Machine learning algorithms have been trained to use patterns of brain activity to "decode" stimuli presented to humans. Here, we demonstrate that the same technique can decode naturalistic video content from the brains of two domestic dogs. We find that decoders based on the actions in the videos were successful in dogs.

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Neuroscience

Intrathecal Vector Delivery in Juvenile Rats via Lumbar Cistern Injection
Anthony Donsante 1, Shauna A. Rasmussen 2, Judith L. Fridovich-Keil 2
1Department of Neurosurgery, School of Medicine, Emory University, 2Department of Human Genetics, School of Medicine, Emory University

A surgical procedure is described to perform injections into the lumbar cistern of the juvenile rat. This approach has been used for the intrathecal delivery of gene therapy vectors, but it is anticipated that this approach can be used for a variety of therapeutics, including cells and drugs.

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Neuroscience

Surgical Lumbar Sympathectomy in Mice
Tina Tian 1,2,3, Patricia J. Ward 2,3
1Medical Scientist Training Program, Emory University School of Medicine, 2Neuroscience Graduate Program, Laney Graduate School, Emory University, 3Department of Cell Biology, Emory University School of Medicine

This manuscript presents a protocol for surgically removing the postganglionic lumbar sympathetic neurons from a mouse. This procedure will facilitate a multitude of studies aimed at investigating the role of sympathetic innervation in distal tissue targets.

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Immunology and Infection

Assessment of Glutamine as a Fuel Source for Alveolar Macrophages Exposed to Chronic Ethanol Using an Extracellular Flux Bioanalyzer
Kathryn M. Crotty 1,2, Samantha M. Yeligar 1,2
1Department of Medicine, Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, Emory University, 2Atlanta Veterans Affairs Health Care System

Alcohol misuse impairs alveolar macrophage (AM) immunity due to suppressed mitochondrial respiration and bioenergetics. We recently demonstrated that ethanol (EtOH) exposure increases glutamine dependency for mitochondrial respiration in AMs. Herein, methods are provided to determine the usage of glutamine for mitochondrial respiration in EtOH-treated AMs using an extracellular flux bioanalyzer.

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