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Whole transcriptome analysis of pancreatic ghrelin cells in WT and Nkx2.2 KO mouse embryo

ABSTRACT: Whole transcriptome analysis of pancreatic ghrelin cells in WT and Nkx2.2 KO mouse embryo

PROVIDER: PRJEB8653 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR769577.fastq.gz	Fastqsanger.gz
	ERR769578.fastq.gz	Fastqsanger.gz
	ERR769579.fastq.gz	Fastqsanger.gz
	ERR769580.fastq.gz	Fastqsanger.gz
	ERR769581.fastq.gz	Fastqsanger.gz

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Whole transcriptome analysis of pancreatic ghrelin cells in WT and Nkx2.2 KO mouse embryo

Project description:Whole transcriptome analysis of Ghrelin-expressing and -descendant cells in pancreas from Nkx2.2 KO and WT mouse embryos.

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Whole ovaries RNA sequencing; WT and ghrelin o-acyltransferase (GOAT) knockout mice

Project description:Ghrelin, an orexigenic gut-derived peptide, is gaining increasing attention due to its multifaceted role in a number of physiological functions, including metabolism, cardiovascular health, stress and reproduction. Ghrelin exists in circulation primarily as des-acylated and acylated ghrelin. Des-acyl ghrelin, until recently considered to be an inactive form ghrelin, is now known to have independent physiological functionality. However, the relative contribution of acyl and des-acyl ghrelin to reproductive development and function is currently unknown. Here we used ghrelin-O-acyltransferase (GOAT) knockout (KO) mice that have no measurable levels of endogenous acyl ghrelin and chronically high levels of des-acyl ghrelin, to characterise how the developmental and life-long absence of acyl ghrelin affects ovarian development and reproductive capacity. We have combined ovarian transcriptome analysis using RNA sequencing with measures of ovarian morphometry, as well as with the assessment of markers of reproductive maturity and the capacity to breed. Our data show pronounced specific changes in the ovarian transcriptome in the juvenile GOAT KO ovary, indicative of advanced ovarian development. These changes corresponded with diminished ovarian reserve in the juvenile and adult ovaries of these mice, due to a continuous reduction in the number of small follicle populations. These changes did not affect the timing of puberty onset or reproductive capacity under optimal conditions. These data suggest that an absence of acyl ghrelin does not prevent reproductive success but that appropriate levels of acyl and des-acyl ghrelin may be necessary for optimal ovarian maturation.

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Ghrelin deletion and conditional ghrelin-cell ablation increase pancreatic islet size in mice

Project description:We utilized single cell sequencing of FACS sorted cells from pancreatic islets of wildtype and ghrelin knock out mice to understand the effects of ghrelin deletion on gene expression profiles of various islet cells.

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Nkx2.2 activates the ghrelin promoter in pancreatic islet cells.

Project description:Nkx2.2 is an essential regulator of pancreatic endocrine differentiation. Nkx2.2-null mice are completely devoid of beta-ells and have a large reduction of alpha- and PP cells. In the place of these islet populations, there is a corresponding increase in the ghrelin-positive epsilon-cells. Molecular studies have indicated that Nkx2.2 functions as an activator and repressor to regulate islet cell fate decisions. To determine whether Nkx2.2 is solely important for islet cell fate decisions or also has the capability to control ghrelin at the promoter level, we studied the transcriptional regulation of the ghrelin promoter within the pancreas, in vitro and in vivo. These studies demonstrate that both of the previously identified transcriptional start sites in the ghrelin promoter are active within the embryonic pancreas; however, the long transcript is preferentially up-regulated in the Nkx2.2-null pancreas. We also show that the promoter region between -619 and -488 bp upstream of the translational start site is necessary for repression of ghrelin in alphaTC1 and betaTC6 cells. Surprisingly, we also show that Nkx2.2 is able to bind to and activate the ghrelin promoter in several cell lines that do or do not express endogenous ghrelin. Together, these results suggest that the up-regulation of ghrelin expression in the Nkx2.2-null mice is not due to loss of repression of the ghrelin promoter in the nonghrelin islet populations. Furthermore, Nkx2.2 may contribute to the activation of ghrelin in mature islet epsilon-cells.

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Gene expression profiling of wild-type, Nkx2.2 null, NeuroD null and Nkx2.2/NeuroD double KO E13.5 pancreata

Project description:The experiment consist of 16 one channel assays for analyzing embryonic pancreas at E13. There are four replicates per condition. We are looking at differential gene expression across four mouse lines: WT, Nkx2.2, NeuroD and Nkx2.2/NeuroD double mutants at E13.5.

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