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California Institute of Technology (Caltech)

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. Henley¹, Bruce N. Cohen¹, Charlene H. Kim¹, Heather D. Gold¹, Rahul Srinivasan¹, Sheri McKinney¹, Purnima Deshpande¹, Henry A. Lester¹

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. Bancroft¹, 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.