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Identification of miRNAs from Glehnia littoralis by high-throughput sequencing and their response to salt stress

ABSTRACT: Glehnia littoralis is a perennial herb growing on the sandy beaches. In this study, high-throughput sequencing of small RNA was performed to identify miRNAs and their response to salt stress in G. littoralis. As a result, 37 conserved miRNAs belonging to 13 families and 37 novel miRNAs were identified. Among the 74 miRNAs, the expression levels of miR-156b*, miR-171, miR-171*, and miR-319 were downregulated and that of miR-399a, miR-399a*, miR-399b, miR-399b*, novel-9 and novel-14* were upregulated under salt stress. Target prediction of salt-responsive miRNAs indicated that these miRNAs exerted a role by regulating specific stress-responsive genes, such as SPLs. However, a lot of target genes of salt-responsive miRNAs were uncharacterized, which implied that there might be some new miRNA-mediated reguatory of salt response in G. littoralis.

ORGANISM(S): Glehnia littoralis

PROVIDER: GSE84621 | GEO | 2016/07/21

SECONDARY ACCESSION(S): PRJNA330675

REPOSITORIES: GEO

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Project description:We tested the hypothesis that a panel of placental mammal-specific miRNAs and their targets play important to establish receptivity to implantation and their dysregulated expression may be a feature in women with early pregnancy loss. Relative expression levels of miR-340-5p, −542-3p, and −671-5p all increased following treatment of Ishikawa cells with progesterone (10 μg/ml) for 24 hrs (p < 0.05). RNA sequencing of these P4-treated cells identified co-ordinate changes to 6,367 transcripts of which 1713 were predicted targets of miR-340-5p, 670 of miR-542-3p, and 618 of miR-671-5p. Quantitative proteomic analysis of Ishikawa cells transfected with mimic or inhibitor (48 hrs: n=3 biological replicates) for each of the P4-regulated miRNAs was carried out to identify targets of these miRNAs. Excluding off target effects, mir-340-5p mimic altered 1,369 proteins while inhibition changed expression of 376 proteins (p < 0.05) of which, 72 were common to both treatments. A total of 280 proteins were identified between predicted (mirDB) and confirmed (in vitro) targets. In total, 171 proteins predicted to be targets by mirDB were altered in vitro by treatment with miR-340-5p mimic or inhibitor and were also altered by treatment of endometrial epithelial cells with P4. In vitro targets of miR-542-3p identified 1,378 proteins altered by mimic while inhibition altered 975 a core of 200 proteins were changed by both. 100 protein targets were predicted and only 46 proteins were P4 regulated. miR-671-mimic altered 1,252 proteins with inhibition changing 492 proteins of which 97 were common to both, 95 were miDB predicted targets and 46 were also P4-regulated. All miRNAs were detected in endometrial biopsies taken from patients during the luteal phase of their cycle, irrespective of prior or future pregnancy outcomes Expression of mir-340-5p showed an overall increase in patients who had previously suffered a miscarriage and had a subsequent miscarriage, as compared to those who had infertility or previous miscarriage and subsequently went on to have a life birth outcome. The regulation of these miRNAs and their protein targets regulate the function of transport and secretion, and adhesion of the endometrial epithelia required for successful implantation in humans. Dysfunction of these miRNAs (and therefore the targets they regulate) may contribute to endometrial-derived recurrent pregnancy loss in women.

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Identification of miRNAs from Glehnia littoralis by high-throughput sequencing and their response to salt stress

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