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Georgia State University

5 ARTICLES PUBLISHED IN JoVE

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Biology

Monitoring ER/SR Calcium Release with the Targeted Ca2+ Sensor CatchER+
Florence N. Reddish 1, Cassandra L. Miller 1, Rakshya Gorkhali 1, Jenny J. Yang 1
1Department of Chemistry, Center of Diagnostics and Therapeutics (CDT), Georgia State University

We present protocols for the application of our targeted genetically-encoded calcium indicator (GECI) CatchER+ for monitoring rapid calcium transients in the endoplasmic/sarcoplasmic reticulum (ER/SR) of HEK293 and skeletal muscle C2C12 cells using real-time fluorescence microscopy. A protocol for the in situ Kd measurement and calibration is also discussed.

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Neuroscience

A High-content Assay for Monitoring AMPA Receptor Trafficking
Mohammad A. Ghane *1, Dina W. Yakout *1, Angela M. Mabb 1
1Neuroscience Institute, Georgia State University

Neuronal excitability can be modulated through a dynamic process of endo- and exocytosis of excitatory ionotropic glutamate receptors. Described here is an accessible, high-content assay for quantifying surface and internal receptor population pools.

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Immunology and Infection

Metal-Limited Growth of Neisseria gonorrhoeae for Characterization of Metal-Responsive Genes and Metal Acquisition from Host Ligands
Stavros Maurakis 1, Cynthia Nau Cornelissen 1
1Institute for Biomedical Sciences, Georgia State University

We describe here a method for growth of Neisseria gonorrhoeae in metal-restricted liquid medium to facilitate the expression of genes for metal uptake. We also outline downstream experiments to characterize the phenotype of gonococci grown in these conditions. These methods may be adapted to be suitable for characterization of metal-responsive genes in other bacteria.

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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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Neuroscience

Limited Bedding and Nesting as a Model for Early-Life Adversity in Mice
Fadya H. Mroue-Ruiz 1, Madison Garvin 1, Logan Ouellette 1, Michelle K. Sequeira 1, Hannah Lichtenstein 1, Urjoshi Kar 1, Jessica L. Bolton 1
1Neuroscience Institute, Georgia State University

This protocol describes an animal model for studying how early-life adversity, provoked by an impoverished environment and unpredictable maternal care during the early postnatal period, affects brain development and the future risk of mental disorders.

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