MicroRNA-432 inhibits milk fat synthesis by targeting <i>SCD</i> and <i>LPL</i> in ovine mammary epithelial cells

Hao, Zhiyun; Luo, Yuzhu; Wang, Jiqing; Hickford, Jon; Zhou, Huitong; Hu, Jiang; Liu, Xiu; Li, Shaobin; Ke, Na; Liang, Weiwei; Huang, Zhaochun

doi:10.1039/d1fo01260f

Cited by 14 publications

(14 citation statements)

References 39 publications

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“…The effect of oar-miR-432 of ovine preadipocytes on the induced differentiation has not been reported previously. It has been shown that miR-432 inhibited milk fat synthesis in sheep mammary epithelial cells ( Hao et al, 2021 ). During myoblast proliferation and differentiation, miR-432 was negatively regulated in pigs ( Ma et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

“…In this study, miRNA-seq and mRNA-seq were used to identify potential miRNAs and mRNA related to sheep fat deposition in Hu fat-tailed and Tibetan thin-tailed sheep breeds. The oar-miR-432 associated with fat synthesis was identified, which is also one of the most downregulated miRNAs ( Hao et al, 2021 ). The TargetScan, miRanda, and RNAhybrid packages of software were used to predict the potential target genes of oar-miR-432 ( John et al, 2004 ; Krüger and Rehmsmeier, 2006 ; Agarwal et al, 2015 ) and putative target genes that overlapped with differentially expressed genes (DEGs).…”

Section: Introductionmentioning

confidence: 99%

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Oar-miR-432 Regulates Fat Differentiation and Promotes the Expression of BMP2 in Ovine Preadipocytes

Jin

Fei

et al. 2022

Front. Genet.

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The fat tail is a unique characteristic of sheep that represents energy reserves and is a complex adaptative mechanism of fat-tailed sheep to environmental stress. MicroRNA plays a significant role as regulators at the posttranscriptional level, but no studies have explained the molecular mechanisms of miRNA which regulate fat deposition in sheep tails. In this study, mRNA and miRNA analysis examined tail fat tissue from three Hu fat-tailed and three Tibetan thin-tailed sheep. After aligning to the reference sequences, 2,108 differentially expressed genes and 105 differential expression miRNAs were identified, including 1,247 up- and 861 downregulated genes and 43 up- and 62 downregulated miRNAs. Among these differentially expressed miRNAs, oar-miR-432 was one of the most downregulated miRNAs between Hu sheep and Tibetan sheep, and 712 genes were predicted to be targeted by oar-miR-432, 80 of which overlapped with DEGs. The Gene Ontology analysis on these genes showed that BMP2, LEP, GRK5, BMP7, and RORC were enriched in fat cell differentiation terms. The genes for BMP2 targeted by oar-miR-432 were examined using dual-luciferase assay. The oar-miR-432 mimic transfected into preadipocytes resulted in increased expression of BMP2. The marker gene PPAR-γ of fat differentiation had a lower expression than the negative control on days 0, 2, and 4 after induced differentiation. The decrease in the number of lipids in the oar-miR-432 mimic group detected by oil red O stain was also less than that in the negative control. This is the first study to reveal the fat mechanisms by which oar-miR-432 inhibits fat differentiation and promotes the expression of BMP2 in sheep tails.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Oar-miR-432 Regulates Fat Differentiation and Promotes the Expression of BMP2 in Ovine Preadipocytes

Jin

Fei

et al. 2022

Front. Genet.

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“…Meanwhile, VLDLR also strengthens the activity of LPL that releases long-chain fatty acids from chylomicron and very low-density lipoprotein. Longchain fatty acids are used as raw materials for synthesizing triglycerides in MECs (13). In addition, knockout mice of VLDLR exhibited a decrease in adipocyte triglyceride storage (30).…”

Section: Discussionmentioning

confidence: 99%

“…The OMECs were isolated according to the method of Hao et al (13). Briefly, cells were cultured in a basal DMEM/F12 medium (Invitrogen, CA, USA) consisting of 10% fetal bovine serum, 5 µg/ml hydrocortisone, 5 µl/ml insulin-transferrinsodium selenium, 10 ng/ml epidermal growth factor 1, and 1 µg/ml hydrocortisone.…”

Section: Animals and Cell Culturementioning

confidence: 99%

MicroRNA-199a-3p regulates proliferation and milk fat synthesis of ovine mammary epithelial cells by targeting VLDLR

Wang

Hao

et al. 2022

Front. Vet. Sci.

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In our previous study, microRNA (miR)-199a-3p was found to be the most upregulated miRNA in mammary gland tissue during the non-lactation period compared with the peak-lactation period. However, there have been no reports describing the function of miR-199a-3p in ovine mammary epithelial cells (OMECs) and the biological mechanisms by which the miRNA affects cell proliferation and milk fat synthesis in sheep. In this study, the effect of miR-199a-3p on viability, proliferation, and milk fat synthesis of OMECs was investigated, and the target relationship of the miRNA with very low-density lipoprotein receptor (VLDLR) was also verified. Transfection with a miR-199a-3p mimic increased the viability of OMECs and the number of Edu-labeled positive OMECs. In contrast, a miR-199-3p inhibitor had the opposite effect with the miR-199a-3p mimic. The expression levels of three marker genes were also regulated by both the miR-199a-3p mimic and miR-199-3p inhibitor in OMECs. Together, these results suggest that miR-199a-3p promotes the viability and proliferation of OMECs. A dual luciferase assay confirmed that miR-199a-3p can target VLDLR by binding to the 3′-untranslated regions (3'UTR) of the gene. Further studies found a negative correlation in the expression of miR-199a-3p with VLDLR. The miR-199a-3p mimic decreased the content of triglycerides, as well as the expression levels of six milk fat synthesis marker genes in OMECs, namely, lipoprotein lipase gene (LPL), acetyl-CoA carboxylase alpha gene (ACACA), fatty acid binding protein 3 gene (FABP3), CD36, stearoyl-CoA desaturase gene (SCD), and fatty acid synthase gene (FASN). The inhibition of miR-199a-3p increased the level of triglycerides and the expression of LPL, ACACA, FABP3, SCD, and FASN in OMECs. These findings suggest that miR-199a-3p inhibited milk fat synthesis of OMECs. This is the first study to reveal the molecular mechanisms by which miR-199a-3p regulates the proliferation and milk fat synthesis of OMECs in sheep.

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“…MicroRNAs (miRNAs) are a class of small, evolutionarily conserved non-coding RNA molecules (~22 nucleotides). They can inhibit translation or promote the degradation of the target mRNAs at a post-transcriptional level by complementarily binding to the 3′-untranslated regions (3′-UTR) of the target genes [ 5 ]. Previous studies have suggested that miRNAs have important roles in regulating the development of SMSCs.…”

Section: Introductionmentioning

confidence: 99%

MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting PTEN and JAG2

Wang

Zhen

Liu

et al. 2022

IJMS

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In our previous study, microRNA (miR)-381 was found to be the most down-regulated miRNA in skeletal muscle of Liaoning cashmere goats with higher skeletal muscle mass, but the molecular mechanism involved remains unclear. In this study, primary caprine skeletal muscle satellite cells (SMSCs) were isolated and identified. We investigated the effect of miR-381 on the viability, proliferation and differentiation of caprine SMSCs, and the target relationships of miR-381 with jagged canonical Notch ligand 2 (JAG2) and phosphatase and tensin homolog (PTEN). Cells isolated were positive for SMSC-specific marker protein Pax7. This suggests that purified SMSCs were obtained. The expression level of miR-381 achieved a peak value on day 4 after SMSC differentiation, and miR-381 also significantly increased the expression levels of myogenic differentiation marker genes: myosin heavy chain (MyHC), myogenin (MyoG) and myocyte enhancer factor 2C (MEF2C) in differentiated SMSCs, the area of MyHC-positive myotubes and the myogenic index. These findings suggest that miR-381 promoted myogenic differentiation of caprine SMSCs. The CCK8 assay and EDU staining analysis showed that miR-381 mimic both inhibited the viability of SMSCs and decreased the percentage of EDU-labeled positive SMSCs. In contrast, miR-381 inhibitor had the opposite effect with miR-381 mimic. A dual luciferase reporter assay verified that miR-381 can target JAG2 and PTEN by binding to the 3′-untranslated regions (3′-UTR) of the genes. The transfection of miR-381 mimic into caprine SMSCs resulted in decreases in expression levels of JAG2 and PTEN, while miR-381 inhibitor increased the two target genes in expression. This is the first study to reveal the biological mechanisms by which miR-381 regulates caprine SMSC activities.

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MicroRNA-432 inhibits milk fat synthesis by targeting SCD and LPL in ovine mammary epithelial cells

Abstract: In our previous studies, microRNA-432 (miR-432) was found to be one of differentially expressed miRNAs in ovine mammary gland between the two breeds of lactating sheep with different milk production...

Cited by 14 publications

References 39 publications

Oar-miR-432 Regulates Fat Differentiation and Promotes the Expression of BMP2 in Ovine Preadipocytes

Oar-miR-432 Regulates Fat Differentiation and Promotes the Expression of BMP2 in Ovine Preadipocytes

MicroRNA-199a-3p regulates proliferation and milk fat synthesis of ovine mammary epithelial cells by targeting VLDLR

MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting PTEN and JAG2

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