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Identifying hand2 downstream targets in cardiomyocyte during early cardiogenesis

ABSTRACT: Identifying hand2 downstream targets in cardiomyocyte during early cardiogenesis

PROVIDER: PRJNA1010830 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25820349_1.fastq.gz	Fastqsanger.gz
	SRR25820349_2.fastq.gz	Fastqsanger.gz
	SRR25820350_1.fastq.gz	Fastqsanger.gz
	SRR25820350_2.fastq.gz	Fastqsanger.gz
	SRR26113048_1.fastq.gz	Fastqsanger.gz

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Identifying hand2 downstream targets in cardiomyocyte during early cardiogenesis

Project description:hand2 reporter-expressing cardiac cells fail to migrate to the midline. To investigate the underlying molecular changes, we sequenced mRNA from 3900 and 3836 hand2 reporter-expressing single cells from 24 hpf hand2 FLD-/- and hand2 FLD+/? sibling embryos, respectively. Furthermore, hand2 is broadly expressed within the LPM, which gives rise to various cell lineages and tissues including the cardiovascular system, blood, kidneys, mesothelium, and limb connective tissue. To further investigate the requirement of Hand2 in the early events of cardiac lineage specification in zebrafish, we aim to characterize the early hand2 expressing cardiac precursors by using embryonic CM H3K27ac ChIP-seq and ATAC-seq data.

2024-08-01 | GSE241971 | GEO

Transcription profile of zebrafish hand2 (hanS6 allele) mutant embryos at 19hpf

Project description:Heart formation requires the fusion of bilateral cardiomyocyte populations as they move toward the embryonic midline. The bHLH transcription factor Hand2 is essential for cardiac fusion; however, the effector genes that execute this function of Hand2 are unknown. Here, we provide the first evidence for a downstream component of the Hand2 pathway that mediates cardiac morphogenesis. Although hand2 is expressed in cardiomyocytes, mosaic analysis demonstrates that hand2 plays a non-autonomous role in regulating cardiomyocyte movement. Gene expression profiles reveal heightened expression of fibronectin 1 (fn1) in hand2 mutant embryos. Reciprocally, overexpression of hand2 leads to decreased Fibronectin levels. Furthermore, reduction of fn1 function enables rescue of cardiac fusion in hand2 mutants: bilateral cardiomyocyte populations merge and exhibit improved tissue architecture, albeit without major changes in apicobasal polarity. Together, our data provide a novel example of a tissue creating a favorable environment for its morphogenesis: the Hand2 pathway establishes an appropriate environment for cardiac fusion through negative modulation of Fn1 levels. Embryos from three independent hand2 mutant (hanS6 allele) heterozyogous crosses were collected. Examination of Tg(myl7:egfp) expression allow sorting of hand2 mutant embryos from their wild-type siblings.

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Transcription profile of zebrafish hand2 (hanS6 allele) mutant embryos at 19hpf

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Gene expression profiling of uterine stromal cells isolated from Hand2 floxed and Hand2 ablated mice on day 4 of pregnancy

Project description:Our previous study revealed that the basic helix-loop-helix transcription factor Hand2 is a downstream target of progesterone signaling in mouse uterine stroma at the time of implantation. Further, conditional deletion of Hand2 in mouse uterus leads to implantation failure due to impaired uterine epithelial receptivity. To identify the downstream targets of Hand2 in the uterus, we performed gene expression profling of uterine stromal cells isolated from Hand2-null mice and the corresponding controls on day4 of pregnancy (the time of implantation). The microarray results revealed elevated expression of mRNAs corresponding to several members of the fibroblast growth factor family in uterine stroma of Hand2-ablated mice. These factors act as paracrine mediators of mitogenic effects of estrogen on the epithleium. Thus, Hand2 is a critical regulator of the uteirne stromal-epithelial communication that directs proper steroid regulation conducive for establshment of pregnancy. We performed conditional ablation of Hand2 in the mouse uterus using the PRcre mouse model. As Hand2 expression is restricted to stromal comaprtment, we isolated uterine stromal cells from day4 pregnant mice (n=5 for each genotype), purified total RNA from these cells, pooled these samples and then hybridized to high density affymetrix microarrays. Control vs. KO.

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