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university of texas medical branch

4 ARTICLES PUBLISHED IN JoVE

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Biology

Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor
Askar M. Akimzhanov 1, Darren Boehning 1
1Department of Neuroscience and Cell Biology, University of Texas Medical Branch

This protocol describes a method for real-time measurement of mitochondrial calcium fluxes by fluorescent imaging. The method takes advantage of a circularly permutated YFP-based dual-excitation ratiometric calcium sensor (ratiometric pericam-mt) selectively expressed in mitochondria.

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Medicine

Purification and Aggregation of the Amyloid Precursor Protein Intracellular Domain
Amina El Ayadi 1, Emily S. Stieren 2, José M. Barral 2, Andres F. Oberhauser 2, Darren Boehning 2
1Department of Surgery, University of Texas Medical Branch , 2Department of Neuroscience and Cell Biology, University of Texas Medical Branch

A method for large-scale purification of the APP intracellular domain (AICD) is described. We also describe methodology to induce in vitro AICD aggregation and visualization by atomic force microscopy. The methods described are useful for biochemical/structural characterization of the AICD and the effects of molecular chaperones on its aggregation.

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Immunology and Infection

In Vivo Imaging Systems (IVIS) Detection of a Neuro-Invasive Encephalitic Virus
Allison Poussard *1, Michael Patterson *1, Katherine Taylor 1, Alexey Seregin 1, Jeanon Smith 1, Jennifer Smith 1, Milagros Salazar 1, Slobodan Paessler 1
1Experimental Pathology, University of Texas Medical Branch

Utilizing luciferase and in vivo imaging systems (IVIS) as a novel means to identify disease endpoints before clinical developments occur. IVIS has allowed us to visualize in real time the invasion of encephalitic viruses over multiple days, providing a more accurate disease model for future study. It has also allowed us to identify the potential protective features of antivirals and vaccines faster than currently utilized animal models. The capability to utilize individual animals over multiple time points ensures reduced animal requirements, costs, and overall morbidity to the animals utilized ensuring a more humane and more scientific means of disease study.

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Immunology and Infection

Confocal Imaging of Double-Stranded RNA and Pattern Recognition Receptors in Negative-Sense RNA Virus Infection
Elizabeth Mateer 1, Slobodan Paessler 1, Cheng Huang 1
1Department of Pathology and Institute for Human Infections and Immunity, University of Texas Medical Branch

Double-stranded RNA produced during RNA virus replication can be recognized by pattern recognition receptors to induce an innate immune response. For negative-sense RNA viruses, the interaction between the low-level dsRNA and PRRs remains unclear. We have developed a confocal microscopy method to visualize arenavirus dsRNA and PRR in individual cells.

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