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Distinct roles of Hand2 in developing and adult autonomic neurons

ABSTRACT: Distinct roles of Hand2 in developing and adult autonomic neurons

PROVIDER: PRJNA419955 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6325330.fastq.gz	Fastqsanger.gz
	SRR6325327.fastq.gz	Fastqsanger.gz
	SRR6325328.fastq.gz	Fastqsanger.gz
	SRR6325329.fastq.gz	Fastqsanger.gz

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Distinct roles of Hand2 in developing and adult autonomic neurons

Project description:Purpose: Analyze the function of the transcription factors Hand2 and Gata3 in adult sympathetic neurons by induced knockout and RNAseq analysis

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Genome-wide candidate HAND2 target regions in mouse embryonic tissues

Project description:The bHLH transcriptional regulator HAND2 performs essential functions during development of the embryonic limb, heart and branchial arches, however, its genomic target regions remained largely unkown. Here, we uncover the range of candidate HAND2 target cis-regulatory modules in the major Hand2 expressing embryonic tissues during midgestation and focus on the transcriptional networks controlled by HAND2 in the developing limb. Our functional and genetic analysis reveals that in the early limb mesenchyme HAND2 directly regulates the expression of several transcription factors involved in the morphogenesis of limb proximal skeletal elements. In addition, HAND2 operates as a key factor in the establishment of anterior and posterior compartments, partly through direct binding to a Gli3 cis-regulatory module. Furthermore, HAND2 refines the posterior boundary of Gli3 expression domain through a relay mechanism mediated by Tbx3. These HAND2-dependent regulatory interactions are required to set the boundaries defining the localized expression of the Shh organizer region, which allows progression of distal limb outgrowth.

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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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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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