|Application:||Gene expression microarray analysis|
|Number of samples:||8|
|Release date:||Jan 31 2016|
|Last update date:||May 11 2017|
|Dataset link||Expression signatures of dopaminergic neurons of ventral SNc (Pitx3-dependent), dorsal SNc (Pitx3-independent) and VTA|
Ventral midbrains were dissected from Pitx3+/+;TH-EGFP+/- and Pitx3-/-;TH-EGFP+/- mouse pups (P1-P4). SNc and VTA were separated for both mouse genotypes (4 sample types: VTA WT, VTA KO, SNc WT and SNc KO) and then tissues were digested in papain solution. Propidium-iodide (PI) was then added to the single cell suspensions and EGFP+/PI- mDA neurons were sorted by flow cytometry into RNAlater solution (3000 cells per 30uL). Batches of EGFP+ cells in RNAlater were stored at -80°C until the necessary number of cells had been accumulated for enough total RNA (100ng) per sample type, as recommended by Affymetrix and Nugen companies. Prior to purification of total RNA, corresponding batches of cells were thawed from -80°C and pooled together in order to achieve the required amount of total RNA for each sample type (VTA WT, VTA KO, SNc WT and SNc KO) in duplicate (8 samples in total).
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