Texas A&M Institute for Neuroscience
2 ARTICLES PUBLISHED IN JoVE
Neuroscience
Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression
Beverley M. Henley 1, Bruce N. Cohen 1, Charlene H. Kim 1, Heather D. Gold 1, Rahul Srinivasan 1, Sheri McKinney 1, Purnima Deshpande 1, Henry A. Lester 1
1Division of Biology and Biological Engineering, California Institute of Technology (Caltech)
In Parkinson's Disease (PD), Substantia Nigra (SNc) dopaminergic neurons degenerate, leading to motor dysfunction. Here we report a protocol for culturing ventral midbrain neurons from a mouse expressing eGFP driven by a Tyrosine Hydroxylase (TH) promoter sequence, harvesting individual fluorescent neurons from the cultures, and measuring their transcriptome using RNA-seq.
Neuroscience
Quantifying Spontaneous Ca2+ Fluxes and their Downstream Effects in Primary Mouse Midbrain Neurons
Eric A. Bancroft 1, Rahul Srinivasan 1,2
1Department of Neuroscience and Experimental Therapeutics, Texas A&M University Health Science Center, College of Medicine, 2Texas A&M Institute for Neuroscience
Here we present a protocol to measure in vitro Ca2+ fluxes in midbrain neurons and their downstream effects on caspase-3 using primary mouse midbrain cultures. This model can be employed to study pathophysiologic changes related to abnormal Ca2+ activity in midbrain neurons, and to screen novel therapeutics for anti-apoptotic properties.
