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Positional cues underlie cell fate specification during branching morphogenesis of the embryonic mammary epithelium

ABSTRACT: Positional cues underlie cell fate specification during branching morphogenesis of the embryonic mammary epithelium

PROVIDER: PRJNA866373 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR20853900_1.fastq.gz	Fastqsanger.gz
	SRR20853900_2.fastq.gz	Fastqsanger.gz
	SRR20853901_1.fastq.gz	Fastqsanger.gz
	SRR20853901_2.fastq.gz	Fastqsanger.gz
	SRR20853902_1.fastq.gz	Fastqsanger.gz

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Positional cues underlie cell fate specification during branching morphogenesis of the embryonic mammary epithelium

Project description:By single-cell RNA-sequencing across 4 stages of embryonic development, we reconstructed the differentiation trajectories of multipotent mammary stem cells towards basal and luminal fate. Our data revealed that MaSCs can already be resolved into distinct populations exhibiting lineage commitment at the time coinciding with the first sprouting events of mammary branching morphogenesis (E15.5). Through an interactive web tool for the visualization of gene expression across our developmental atlas, we provide novel molecular markers for committed and multipotent MaSCs, and define their spatial distribution within the developing tissue. Furthermore, we show that the mammary embryonic mesenchyme is composed of two spatially-restricted cell populations, representing the sub-epithelial and dermal mesenchyme.

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Positional Identity of Branching-associated HOXB2 and B3 Gene Expression is Maintained by Adult Human Airway Epithelial Progenitor Cells

Project description:In the context that HOX mutant mice have profound deficits in airway branching morphogenesis, we asked whether the positional identity of HOX genes is maintained in the epithelium of the adult human tracheobronchial tree and if so, whether these positional differences are embedded in the stem/progenitor population derived from nonbranching vs branching airways. We found that 10 of 39 HOX genes are expressed in varying degrees in the nonbranching (trachea) or branching (large and small airways) human airway epithelium. Strikingly, HOXB2 and B3 were highly expressed in the trachea epithelium, but barely detectable in the branching airway epithelium. This difference in HOXB2 and B3 gene expression was embedded in the airway basal cell stem/progenitor population isolated from the trachea vs branching airways, and maintained during differentiation in vitro. Finally, HOXB2 and B3 expressions in the trachea vs branching airways correlated with the expression of a variety of other transcription and growth factors related to branching morphogenesis. The finding that the adult human airway epithelium expresses some HOX genes in a positional manner, with differences embedded in the transcriptomes of airway epithelial stem/progenitor cells, has implications for the use of stem/progenitor cells for applications relevant to lung regeneration.

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Time course gene expression profiling of mouse mammary gland TEB and duct cells throughout branching morphogenesis

Project description:Branching morphogenesis of the mammary gland is driven by the highly motile terminal end bud (TEB) throughout pubertal development. The stem cell enriched, proliferative TEB branches as it invades the mammary fat pad to create a complex network of ducts. The gene expression programs specific to the TEB and the differentiated duct are poorly understood. We conducted a time course analysis of gene expression in the TEB and duct throughout branching morphogenesis. Additionally, we determined the gene regulatory networks coordinated by the Co-factor of LIM domains (CLIM/LDB) transcriptional regulators and determined an essential function for CLIMs in branching morphogenesis by maintaining basal mammary epithelial stem cells and promoting cell proliferation. We used laser capture microdissection to isolate TEB and duct cells throughout branching morphogenesis. We then profiled gene expression in these cells to determine gene regulatory networks involved in branching morphogenesis, and specifically those regulated by CLIM transcriptional regulators. Mouse mammary glands from 4, 6, 8, and 10 week old mice (early puberty through early adulthood) were used for laser capture microdissection of TEB and duct cells from WT and K14-DN-Clim transgenic mice. RNA was isolated (Qiagen) and hybridized to Affymetrix MouseGene 1.0 ST arrays. In addition, basal (CD29HiCD24+Lin-) and Luminal (CD29LoCD24+Lin-) cells were sorted and RNA collected for hybridization to Affymetrix MouseGene 1.0ST arrays.

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Time course gene expression profiling of mouse mammary gland TEB and duct cells throughout branching morphogenesis

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