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| Status |
Public on Nov 28, 2022 |
| Title |
Establishment of a dopaminergic neuron purification system in mice |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Transcriptomic analysis of enriched of neuron population collected from in-vivo mouse brains has been a challenge in the neuroscience field due to its fragility in withstanding harsh condition during isolation and collection process. We established a fluorescent reporter mouse Ddc-hKO1 to facilitate the identification and collection of Ddc-expressing neurons (dopaminergic, serotonergic, cholinergic and adrenergic neurons) by the detection of red fluorescence signals using a FACs. We utilized an improved isolation protocol that ensure high yield and quality of viable neurons during collection that is suitable for RNA-sequencing. This is the first report of transcriptomic profiling of Ddc expressing (hKO1(+)) neurons. Successful collection of Ddc-expressing neurons were verified by gene markers of dopaminergic, serotonergic, cholinergic neurons in hKO1(+) populations while other neuron types in hKO1(-) population. Furthermore, GSEA analysis were performed on both hKO1(+) and hKO1(-) to further support the neuron types collected in respective population.
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| Overall design |
mRNA profiles of Ddc-hKO1-positive and Ddc-hKO1-negative neuron cells from around 20,000 cells were used for sequencing.
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| Contributor(s) |
Sheng K, Yamaguchi S, Nakano T |
| Citation(s) |
36568970 |
| |
| Submission date |
Apr 06, 2021 |
| Last update date |
Jan 11, 2023 |
| Contact name |
Kit-yeng Sheng |
| E-mail(s) |
ms.samanthasky@gmail.com
|
| Organization name |
Osaka University
|
| Lab |
Stem cell pathology
|
| Street address |
Yamadaoka 2-2
|
| City |
Suita |
| ZIP/Postal code |
565-0871 |
| Country |
Japan |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA720130 |
| SRA |
SRP313693 |
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