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Keck School of Medicine, University of Southern California

7 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Mosaic Analysis of Gene Function in Postnatal Mouse Brain Development by Using Virus-based Cre Recombination
Daniel A. Gibson 1,2, Le Ma 2,3
1Neuroscience Graduate Program, Keck School of Medicine, University of Southern California, 2Zilkha Neurogenetic Institute, University of Southern California, 3Department of Cell and Neurobiology, Neuroscience Graduate Program, Keck School of Medicine, University of Southern California

An in vivo method to test gene function in postnatal brain is described. Recombinant AAVs expressing Cre and/or a fluorescent protein are injected into neonatal mouse brain. Mosaic gene inactivation and sparse neuronal labeling are achieved, allowing rapid analysis of gene function in processes critical to neural circuit development.

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Immunology and Infection

Dissecting Host-virus Interaction in Lytic Replication of a Model Herpesvirus
Xiaonan Dong 1, Pinghui Feng 2
1Center for Autophagy Research, Department of Internal Medicine, UT Southwestern Medical Center, 2Department of Microbiology, UT Southwestern Medical Center

We describe a protocol to identify key roles of host signaling molecules in lytic replication of a model herpesvirus, gamma herpesvirus 68 (γHV68). Utilizing genetically modified mouse strains and embryonic fibroblasts for γHV68 lytic replication, the protocol permits both phenotypic characterization and molecular interrogation of virus-host interactions in viral lytic replication.

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Medicine

Modeling Colitis-Associated Cancer with Azoxymethane (AOM) and Dextran Sulfate Sodium (DSS)
Ameet I. Thaker 1, Anisa Shaker 1, M. Suprada Rao 1, Matthew A. Ciorba 1
1Division of Gastroenterology, Washington University School of Medicine

We demonstrate a protocol in which administration of the genotoxic agent azoxymethane (AOM) followed by three cycles of the pro-inflammatory agent dextran sulfate sodium (DSS) rapidly and consistently generates colon tumors in mice with morphologic and molecular similarities to those seen in human colitis-associated cancer.

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JoVE Journal

Use of LysoTracker to Detect Programmed Cell Death in Embryos and Differentiating Embryonic Stem Cells
Jennifer L. Fogel 1, Thu Zan Tun Thein 1, Francesca V. Mariani 1
1Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California

We present a simple protocol to visualize regions of programmed cell death (PCD) in mouse embryos and differentiating embryonic stem (ES) cell cultures using a highly soluble dye called LysoTracker.

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Immunology and Infection

Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production
Junjie Zhang 1, Lining Zhu 1, Pinghui Feng 1
1Department of Microbiology and Immunology, Keck School of Medicine, University of Southern California

We describe a protocol to measure the antiviral cytokine production in mice infected with a model herpesvirus, murine gamma herpesvirus 68 (γHV68) that is closely-related to human Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV). Utilizing genetically modified mouse strains and mouse embryonic fibroblasts (MEFs), we assessed the antiviral cytokine production both in vivo and ex vivo. “Reconstituting” the expression of innate immune components in knockout embryonic fibroblasts by lentiviral transduction, we further pinpoint specific innate immune molecules and dissect the key signaling events that differentially regulate the antiviral cytokine production.

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Biology

Isolation of Myofibroblasts from Mouse and Human Esophagus
Matthew Gargus 1, Chao Niu 1, Anisa Shaker 1
1Department of Medicine, Keck School of Medicine, University of Southern California

We present a protocol to generate primary cultures of murine and human esophageal stromal cells with a myofibroblast phenotype. Cultured cells have spindle shaped morphology, express α-SMA and vimentin, and lack epithelial, hematopoietic and endothelial cell surface markers. Characterized stromal cells can be used in functional studies of epithelial-stromal interactions.

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Medicine

A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair
Nikita Tripuraneni *1, Marissa K. Srour *1, John W. Funnell 1, Thu Zan Tun Thein 1, Francesca V. Mariani 1
1Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California

The overall goal of this procedure is to successfully resect a portion of bone from the rib of a mouse. The procedure was developed as a model to study large-scale long bone repair.

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