Minneapolis Veterans Affairs Health Care System

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

Tammy A. Butterick^1,2, Cayla M. Duffy^1,2, Rachel E. Lee³, Charles J. Billington^1,2,4, Catherine M. Kotz^1,2, Joshua P. Nixon^1,2

¹Department of Veterans Affairs, Minneapolis Veterans Affairs Health Care System, ²Department of Food Science and Nutrition, University of Minnesota, ³Department of Integrative Biology and Physiology, University of Minnesota, ⁴Department of Medicine, University of Minnesota Medical School, University of Minnesota

Multiplex assays can provide beneficial information for basic cellular mechanisms and eliminate waste of reagents and unnecessary repetitive experiments. We describe here a multiplex caspase-3/7 activity assay, using fluorescent- and luminescent-based methods, to determine cell viability in an in vitro hypothalamic model following oxidative challenge with palmitic acid.

Neuroscience

Microglia as a Surrogate Biosensor to Determine Nanoparticle Neurotoxicity

Cayla M. Duffy^1,2, Shihab Ahmed¹, Ce Yuan^1,2,3, Vijayakumar Mavanji¹, Joshua P. Nixon^1,2, Tammy Butterick^1,2,4

¹Minneapolis Veterans Affairs Health Care System, ²Department of Food Science and Nutrition, University of Minnesota, ³Biomedical Informatics and Computational Biology Program, University of Minnesota, ⁴Minnesota Obesity Center, University of Minnesota

Microglia (immune cells of the brain), are used as a surrogate biosensor to determine how nanoparticles influence neurotoxicity. We describe a series of experiments designed to assay microglial response to nanoparticles and exposure of hypothalamic neurons to supernatant from activated microglia to determine neurotoxicity.