Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Dicer deficiency reveals microRNAs predicted to control gene expression in developing adrenal cortex [array]

ABSTRACT: MicroRNAs (miRNAs) are small, endogenous, non-protein coding RNAs that are an important means of post-transcriptional gene regulation. Deletion of Dicer, a key miRNA processing enzyme, is embryonic lethal in mice, and tissue-specific Dicer deletion results in developmental defects. Using a conditional knockout model, we generated mice lacking Dicer in the adrenal cortex. These Dicer knockout (KO) mice exhibited perinatal mortality and failure of the adrenal cortex during late gestation between embryonic day 16.5 (E16.5) and E18.5. Further study of Dicer KO adrenals demonstrated a significant loss of Sf1 expressing cortical cells that was histologically evident as early as E16.5 coincident with an increase in p21 and cleaved-caspase 3 staining in the cortex. However, peripheral cortical proliferation persisted in KO adrenals as assessed by anti-PCNA staining. To further characterize the embryonic adrenals from Dicer KO mice, we performed microarray analyses for both gene expression and miRNA on purified RNA isolated from control and KO adrenals of E15.5 and E16.5 embryos. Consistent with the absence of Dicer and the associated loss of miRNA-mediated mRNA degradation, we observed an up-regulation of a small subset of adrenal transcripts in Dicer KO mice, most notably the transcripts coded by the genes Nr6a1 and Acvr1c. Indeed, several miRNAs, including let-7, miR-34c, and miR-21 that are predicted to target these genes for degradation, were also markedly down-regulated in Dicer KO adrenals. Together these data suggest a role for miRNA mediated regulation of a subset of genes that are essential for normal adrenal growth and homeostasis. Adrenals from control and Dicer KO litter mates were pooled separately from 4 individual litters, resulting in a total of 4 control (cre-) and 4 Dicer KO biological replicates at both E15.5 and E16.5.

ORGANISM(S): Mus musculus

SUBMITTER: Katherine Gurdziel

PROVIDER: E-GEOD-45592 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Purpose: The miRNA profiles were compared among groups of chronic stress exposure and control in two different breeds to identify the differentially expressed miRNAs related to response to chronic stress in the pituitary-adrenal axis. Methods: 6 male adult CFD and 6 Beagles were chosen at random with the similarities in good health, weight and other aspects. Separately, 3 of these two breeds were freely selected for the stress exposure via intermittent electrical stimulation and restraint stress, while the other 3 of these two breeds were non-disposed for normal control.The details for the disposal of dogs were: every morning dogs were restrained and electrical stimulations were exerted with a stable current of 10 mA for 6 s and then with a 6 s interval, lasting for 20 min every day. The duration of disposal was ten days.ll 12 dogs were killed by air embolism in the 11th day. Subsequently, pituitary and adrenal cortex tissues were fast collected and isolated for further high-sequencing. Results: 8 small miRNA libraries were constructed for miRNA-seq. A number of reads ranging from 15,926,611 to 81,028,035 was obtained in those 8 groups.a total of 29 (7 up, 22 down), 37 (16 up, 21 down), 30 (22 up, 8 down), 23 (16 up, 7 down), 21 (15 up, 6 down), 50 (11 up, 39 down), 28 (6 up, 22 down) and 40 (29 up, 11 down) DE-miRNAs (P-value < 0.001, log2(Fold_change) >1) were detected in the comparisons of BAC1_vs_BAC2, BAC1_vs_CFDAC1, BAC2_vs_CFDAC2, CFDAC1_vs_CFDAC2, BP1_vs_BP2, BP1_vs_CFDP1, BP2_vs_CFDP2 and CFDP1_vs_CFDP2, respectively Conclusions: Our results shed light on the miRNA profiles with and within chronic stress exposure in adrenal cortex and pituitary. Adrenal cortex and pituitary miRNA profiles of adult Chinese Field Dog and Beagle under chronic stress exposure and normal control, including BAC1 (Beagle adrenal cortex with disposal), BAC2 (Beagle adrenal cortex with non-disposal), BP1 (Beagle pituitary with disposal), BP2 (Beagle pituitary with non-disposal), CFDAC1 (Chinese Field Dog adrenal cortex with disposal), CFDAC2 (Chinese Field Dog adrenal cortex with non-disposal), CFDP1 (Chinese Field Dog pituitary with disposal), CFDP2 (Chinese Field Dog pituitary with non-disposal), were generated by deep sequencing, using Illumina Hiseq 2000.

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Dicer deficiency reveals microRNAs predicted to control gene expression in developing adrenal cortex [array]

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