Primary Culture of Mouse Dopaminergic Neurons

Florence Gaven; Philippe Marin; Sylvie Claeysen

doi:10.3791/51751

Primary Culture of Mouse Dopaminergic Neurons

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Primary Culture of Mouse Dopaminergic Neurons

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39,565 Views

11:58 min

September 8, 2014

DOI: 10.3791/51751-v

Florence Gaven^1,2,3, Philippe Marin^1,2,3, Sylvie Claeysen^1,2,3

¹CNRS, UMR-5203,Institut de Génomique Fonctionnelle, Montpellier, ²Inserm,U661, Montpellier, ³Universités de Montpellier

Dopaminergic neurons play a vital regulatory role in the brain. Their loss is associated with Parkinson's disease. In this video, we show how to generate primary cultures of central dopaminergic neurons from embryonic mouse mesencephalon. Such cultures are useful to study the extreme vulnerability of these neurons to various stresses.

The overall goal of this procedure is to show how to generate a primary culture of dopaminergic neurons from embryonic mouse brains. This is accomplished by first collecting mouse E 13.5 embryos. The second step is to dissect the embryonic mouse brains and to isolate the mesencephalon.

Next, an enzymatic and a mechanical dissociation are applied to the collected mesencephalon. The final step is the plating of dissociated neuronal cells. Ultimately, immunofluorescent staining is used to show the presence of dopaminergic neurons in the culture.

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