A method for isolating cortical interneurons sharing the same birthdays for gene expression studies

Ng, Hui Xuan; Lee, Ean Phing; Cavanagh, Brenton; Britto, Joanne M.; Tan, Seong‐Seng

doi:10.1016/j.expneurol.2017.05.006

Cited by 2 publications

(2 citation statements)

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“…It is well known that Cu (II) is toxic for the cells, as it reduces mRNA transcription and promotes RNA degradation (Halliwell and Gutteridget, 1984; Halliwell et al, 1992). However, there is evidence that, in fixed cells, an 85% reduction in Cu (II) concentration was able to replicate the standard EdU detection capacity (Ng et al, 2017). Hence, we decided to analyze by FC chick brain birthdated samples after modifying Cu (II) concentration in the EdU cocktail ( Figure 3 ).…”

Section: Resultsmentioning

confidence: 99%

“…As mentioned above, Cu (II) can interfere with RNA expression (Halliwell and Gutteridget, 1984; Halliwell et al, 1992; Ng et al, 2017) Consequently, we decided to examine if the new EdU cocktail (containing low Cu (II) concentration) was altering cell gene expression. For that, we purified by FACS a non-revealed control population and a treated population, and compared their gene marker expression by RT-qPCR (Fig.…”

Section: Resultsmentioning

confidence: 99%

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BirthSeq, a new method to isolate and analyze dated cells from any tissue in vertebrates

Rueda-Alaña,

Grillo,

Vazquez

et al. 2023

Preprint

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Embryonic development is a complex and dynamic process that unfolds over time and involves the production of increasing numbers of cells, as well as the diversification of different cell types. The impact of developmental time on the formation of the central nervous system is well documented, with evidence showing that time plays a critical role in establishing the identity of neuronal subtypes. However, the study of how time translates into genetic instructions driving cell fate is limited by the scarcity of suitable experimental tools. We introduce BirthSeq, a new method for isolating and analyzing cells based on their birth date. This innovative technique allows for in vivo labeling of cells, isolation via FACS, and analysis using high-throughput techniques. We demonstrate the effectiveness of BirthSeq for single-cell RNA sequencing and novel spatially resolved transcriptomic approaches in brain development across three vertebrate species (mouse, chick, and gecko). Overall, BirthSeq provides a versatile tool for studying any tissue in any vertebrate organism, helping to fill the necessity in developmental biology research by targeting cells and their temporal cues.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

BirthSeq, a new method to isolate and analyze dated cells from any tissue in vertebrates

Rueda-Alaña,

Grillo,

Vazquez

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

BirthSeq, a new method to isolate and analyze dated cells in different vertebrates

Rueda-Alaña,

Grillo,

Vázquez

et al. 2024

Development

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Embryonic development is a complex and dynamic process that unfolds over time and involves the production and diversification of increasing numbers of cells. The impact of developmental time on the formation of the central nervous system is well-documented, with evidence showing that time plays a critical role in establishing the identity of neuronal subtypes. However, the study of how time translates into genetic instructions driving cell fate is limited by the scarcity of suitable experimental tools. We introduce BirthSeq, a new method for isolating and analyzing cells based on their birth date. This innovative technique allows for in vivo labeling of cells, isolation via FACS, and analysis using high-throughput techniques. We tuned up BirthSeq in developmental organs across three vertebrate species (mouse, chick, and gecko), and fully made use of it for single-cell RNA sequencing and novel spatially resolved transcriptomic approaches in mouse and chick, respectively. Overall, BirthSeq provides a versatile tool for studying virtually any tissue in different vertebrate organism, helping to fill the necessity in developmental biology research by targeting cells and their temporal cues.

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A method for isolating cortical interneurons sharing the same birthdays for gene expression studies

Cited by 2 publications

References 53 publications

BirthSeq, a new method to isolate and analyze dated cells from any tissue in vertebrates

BirthSeq, a new method to isolate and analyze dated cells from any tissue in vertebrates

BirthSeq, a new method to isolate and analyze dated cells in different vertebrates

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