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11 publications mentioning bta-mir-130b

Open access articles that are associated with the species Bos taurus and mention the gene name mir-130b. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

1
[+] score: 572
In addition, the expression analysis of the miR-130b target genes in blastocysts derived from zygotes injected with inhibitor indicated that both the SMAD5 and MSK1 genes were tended to be altered in inhibitor and precursor injected zygote group suggesting that miR-130b could be involving in morula and blastocyst formation by regulating the expression level of SMAD5 and MSK1. [score:12]
Nevertheless, the expression level of miR-130b was not altered in inhibitor transfected group as miRNA inhibitors are not necessary to reduce the expression pattern of the miRNA rather, synthetic miRNA inhibitor are designs to be bind the endogenous mature miRNAs to sequester the endogenous miRNA making it unavailable for normal function. [score:11]
Moreover, in the current study, precursor induced overexpression of miR-130b inhibited the expression (both mRNA and protein) level of MSK1 and SMAD5 gene whereas inhibition of miR-130b increased the protein expression levels of these genes in both cell types. [score:11]
Afterwards, sub confluent cumulus cells were co -transfected with 800 ng/ml of pmirGLO Dual-Luciferase miRNA target reporter vector harboring of the 3′ UTR of the target genes or a mismatch of the target gene and 50 pmole/ml of miR-130b precursor, inhibitor or mismatch control using Lipofectamine 2000 transfection reagent (Invitrogen) in Opti-MEM medium I reduced serum Media. [score:9]
WB, western blot, KDa; kilo Dalton Once, we realized that miR-130b and its target genes were altered in cumulus and granulosa cells transfected with miR-130b precursor or inhibitor, the effect of overexpression or inhibition of miR-130b on granulosa and cumulus cells survival and viability have been examined 24 and 48 h post transfection. [score:9]
WB, western blot, KDa; kilo Dalton Once, we realized that miR-130b and its target genes were altered in cumulus and granulosa cells transfected with miR-130b precursor or inhibitor, the effect of overexpression or inhibition of miR-130b on granulosa and cumulus cells survival and viability have been examined 24 and 48 h post transfection. [score:9]
Following this, the potential role of miR-130b in preimplantation embryo development was investigated by inhibiting and overexpressing its expression by microinjecting the miR-130b inhibitor and miR-130b precursor in the zygote stage embryos, respectively. [score:8]
In the second phase of the study, the role of the miR-130b in granulosa and cumulus cell function were investigated by overexpression or inhibition of miR-130b expression by transfecting the cumulus or granulosa cells with miR-130b precursor or inhibitor while the role of miR-130b on oocyte maturation and preimplantation embryo development was investigated by microinjecting the GV stage oocyte and zygote, respectively with miR-130b precursor or inhibitor. [score:8]
The role of miR-130b in granulosa and cumulus cell function was investigated by increasing and inhibiting its expression in in vitro cultured cells using miR-130b precursor and inhibitor, respectively while the role of miR-130b on oocyte development, immature oocytes were microinjected with miR-130b precursor and inhibitor and the polar body extrusion, the proportion of oocytes reaching to metaphase II stage and the mitochondrial were determined in each oocyte group 22 h after microinjection. [score:8]
The signal transduction pathways regulated by miR-130b during the oocyte maturation is not clear: However, when we look into the expression of pattern of the target genes, the expression level of both SMAD5 and MSK1 were significantly increased in oocytes with lower maturation rate suggesting that miR-130b could be involved in oocyte maturation by fine-tuning the SMAD5 and MSK1 genes. [score:8]
Furthermore, inhibition of miR-130b expression during oocyte in vitro maturation reduced the first polar body extrusion, the proportion of oocytes reaching to metaphase II stage and the mitochondrial activity, while inhibition of miR-130b during preimplantation embryo development significantly reduced morula and blastocyst formation. [score:8]
Similar to our findings, overexpression of miR-130b was found to increase the proliferation rates of the esophageal squamous cell carcinoma and U251 cells while suppression of its expression reduced the proliferation rate of both cell types [45, 46]. [score:7]
Fig. 6The expression levels of miR-130b and its target genes in cumulus (a) and granulosa (b) cells transfected with 130b precursor, miR-130 inhibitor and scramble miRNA 24 h post transfection. [score:7]
The firefly and renilla activity ratio in cells transfected only with vector construct harboring the 3′ UTR of the miR-130b target genes or cells co -transfected along with the miRNA scramble sequence or the firefly and renilla activity ratio of cells co -transfected with miR-130b precursor or inhibitor along with the vector construct harboring the non-target sequence of miR-130b were used as controls. [score:7]
Here we have shown that in vitro functional modulation of miR-130b using its mimic and inhibitor affected the granulosa and cumulus cell function, oocyte maturation and preimplantation embryo development by targeting SMAD5 and MSK1 genes suggesting that miR-130b is involved in bovine oocyte and preimplantation embryo development. [score:7]
Furthermore, down-regulation of miR-130b significantly suppressed cell proliferation and induce apoptosis in HL-60 cells [47] suggesting the potential role of miR-130b in cell survival and proliferation. [score:6]
On the other hand, the luciferase activity was increased in the cumulus cells co -transfected with miR-130b inhibitor and SMAD5GlO compared to cells co -transfected with scrambled miRNA and SMAD5GlO or miR-130b precursor and pmiRGLO harboring the 3′ UTR sequence of the non target sequence (Non-targetGLO) or the cumulus cells transfected only with SMAD5Glo (Fig. 4a). [score:6]
b The luciferase reporter assay in cumulus cells co -transfected with miR-130b precursor or inhibitor and the pmirGLO vector construct harboring of the 3′ UTRs of the MSK1 gene (MSK1GLO) or non-target sequence (Non-targetGLO). [score:6]
In line to this, our results clearly showed that while overexpression of miR-130b increased, inhibition reduced the lactate production suggesting that miR-130b is involving in maintaining cumulus cell survival and proliferation by regulating the glycolysis activity. [score:6]
The expression miR-130b was significantly increased in the blastocysts derived from miR-130b precursor injected zygotes (Fig. 11a) and the mRNA and protein expression of SMAD5 and MSK1 tended to be higher and lower in blastocysts derived from zygotes injected with inhibitor and precursor, respectively compared to blastocysts derived from zygotes injected with scramble or uninjected groups (Fig. 11b– d). [score:6]
a The luciferase reporter assay in cumulus cells co -transfected with miR-130b precursor or inhibitor and the pmirGLO vector construct harboring of the 3′ UTRs of the SMAD5 gene (SMAD5GLO) or non-target sequence (Non-targetGLO). [score:6]
The protein expression level of MSK1 and SMAD5 in miR-130b precursor, inhibitor and scrambled miRNA injected oocytes 24 h post injection (d). [score:5]
Following this, 2-cell (n = 100), 4-cell (n = 75), 8-cell (n = 50), morula (n = 50) and blastocyst (n = 50) stage embryo were collected for miR-130b and its target genes expression analysis. [score:5]
In the current study, inhibition of miR-130b during oocyte maturation significantly affected the oocyte maturation rate, but overexpression had only minimal effect. [score:5]
Following this, the expression profiles of miR-130b and its target genes in MII oocytes derived from different treatment groups were analyzed. [score:5]
The red color indicates the expression level of miR-130b, U6 or scramble miRNA probe while the blue color indicates the nuclear staining using 4′,6-diamidino-2-phenylindole (DAPI) Fig. 3 Whole-mount in situ detection of miR-130b expression in oocytes (immature COCs, mature COCs) and preimplantation embryos. [score:5]
GV Germinal vesicle, SD standard deviation Fig. 9The expression level of miR-130b (a), SMAD5 (b) and MSK1(c) in oocytes derived from GV oocytes injected with miR-130b precursor, miR-130b inhibitor and scrambled miRNA 24 h post injection. [score:5]
Significant differences (p < 0.05) are indicated by star (*) symbol Since modulation of miR-130b altered the rate of cumulus cell proliferation, we hypothesized that overexpression or inhibition of miR-130b could affect the cumulus cell function by compromising the energy metabolism. [score:5]
Western blot analysis showing the protein expression level of SMAD5 and MSK1 in blastocyst derived from zygotes of different treatment groups (d) The expression pattern of miR-130b was analyzed in immature and in vitro-matured oocytes, cumulus cells and granulosa cells. [score:5]
Therefore, in this study, the expression analysis of genes targeted by miR-130b was analyzed in each cell culture group. [score:5]
The 3′-UTRs of the selected genes and the non-target sequences were integrated into the pmiRGLO vector and co -transfected along with miR-130b precursor, miR-130b inhibitor, or a scrambled miRNA sequence into the cultured cumulus cells. [score:5]
The result indicated that the expression level of miR-130b was significantly increased (Fig. 9a), whereas the expression levels (both the mRNA and protein) of SMAD5 (Fig. 9b & d) and MSK1 (Fig. 9c & d) were reduced in MII oocytes derived from GV oocytes injected with miR-130b precursor. [score:5]
Fig. 8The effect of miR-130b overexpression or inhibition on cumulus cell glycolytic activity (a) and cholesterol biosynthesis (b). [score:5]
Hpi hours post insemination (fertilization), SD standard deviation Fig. 11The expression levels of miR-130b and its target genes in blastocysts of different groups. [score:5]
However, inhibition of the already accumulated miR-130b using its inhibitor was able to show substantial effect on oocyte maturation. [score:5]
Inhibition of miR-130b expression reduced the morula and blastocyst formation. [score:5]
Overexpression of miR-130b increased the granulosa and cumulus cell proliferation, while inhibition showed the opposite phenotype. [score:5]
Expression analysis of genes targeted by miR-130b in oocyte companion cells, oocytes, and preimplantation embryos. [score:5]
In the current study, overexpression miR-130b promoted granulosa and cumulus cell viability and proliferation; while inhibition of miR-130b resulted in the opposite phenotype. [score:5]
Indeed, overexpression or inhibition of miR-130b didn’t affect the cleavage rate of the zygotes 24 h post injection. [score:5]
Similarly, the expression level of MSK1 tended to be reduced in miR-130b precursor transfected and increased in inhibitor transfected cumulus (Fig. 6a) and granulosa cells (Fig. 6b). [score:5]
However, inhibition of miR-130b during preimplantation embryo development affected the morula and blastocysts formation rates indicating the possible involvement of miR-130b in bovine preimplantation development. [score:5]
Following this, the expression level of miR-130b and its target genes was analyzed in blastocysts derived from the different zygote groups. [score:5]
Total RNA isolation from oocyte granulosa & cumulus cells, oocytes, and preimplantation embryos and complementary DNA synthesis for expression analysis of miR-130b and its target gens. [score:5]
The expression levels of miR-130b and its target genes were also analyzed in blastocysts derived from each zygote group. [score:5]
Fig. 7The effect of miR-130b overexpression or inhibition on cumulus and granulosa cell survival and proliferation. [score:5]
In addition, the expression pattern of the miR-130b was analyzed during bovine preimplantation embryo stages and result showed that miR-130b was expressed in all stages of preimplantation embryos. [score:5]
The expression level of the miR-130b target genes in granulosa cells, immature cumulus, mature cumulus, and all preimplantation stage embryos or different treatment groups were determined using the relative standard curve method according to Larionov et al. [39] and the manual described by Applied Biosystems (http://tools. [score:5]
For this, the expression levels miR-130b was modulated during in vitro oocyte maturation by microinjecting miR-130b precursor or inhibitor into the germinal vesicle (GV) stage oocytes. [score:5]
The miRNA-130b precursors, which mimic endogenous miRNA-130b, miRNA-130b inhibitors that specifically bind to the endogenous miR-130b and scramble miRNA which don’t target any annotated genes were purchased from Ambion (USA). [score:5]
The expression levels of miR-130b and its target genes were also analyzed in the MII-stage oocytes derived from each injected oocyte group. [score:5]
The expression of miR-130b was relatively constant until embryonic stages that coincide with bovine major genome activation and then the expression was increased in morula and blastocyst stages of embryos. [score:5]
Afterwards, 200–500 base pairs flanking the miR-130b binding site and Pme 1 and Xhol restriction sites in the 3′ UTR of the target genes and a gene sequence (mismatch) that is not target by miR-130b were amplified by polymerase chain reaction (PCR) using gene-specific primers (Additional file 1: Table S1). [score:4]
COCs; cumulus oocyte complexes To identify the target genes of miR-130b, in silico predicted target genes (SMAD5, RPS6KA5 (MSK1), MEOX2, DOC1R, MARCH2, DDX6, EIF2C1 and EIF2C4) were experimentally validated using dual luciferase assay. [score:4]
Microinjection of miR-130 precursor or inhibitor indicated that although zygotes of different treatment groups showed no significant differences in the cleavage rates 48 h post fertilization, morula and blastocyst formation were significantly reduced in zygotes injected with miR-130b inhibitor, compared to the scramble injected and uninjected control groups (Table 3). [score:4]
Prior to performing target gene validation using dual luciferase assay, the potential genes targeted by miR-130b were predicted using PicTar (http://www. [score:4]
Thus, to verify whether miR-130b is involving in preimplantation embryo development, bovine zygotes were injected with miR-130b precursor or inhibitor. [score:4]
Zhao G Zhang JG Shi Y Qin Q Liu Y Wang B MiR-130b is a prognostic marker and inhibits cell proliferation and invasion in pancreatic cancer through targeting STAT3PLoS One. [score:4]
However, a better understanding of miR-130b using a stable knockdown or knock-in experiments could be required to get an in-depth insight about the role of miR-130b in the later stage of embryos development, particularly during the period of embryo implantation. [score:4]
The lesser effect of overexpression of miR-130b on oocyte maturation could be associated with its abundance level of which its expression was stronger in the GV oocytes compared to the MII ones. [score:4]
Prior to functional analysis, the expression profile of miR-130b was analyzed in granulosa cells, immature oocytes and corresponding cumulus cells, matured oocyte and corresponding cumulus cells and preimplantation embryos. [score:3]
The expression level of zygotes was used as a reference sample to calculate the fold change expression of miR-130b between preimplantation stage embryos. [score:3]
With respect to this, previously, we identified altered expression of microRNA-130b (miR-130b) during oocyte maturation. [score:3]
However, the expression level of miR-130b was progressively increased in the morula and blastocyst embryonic stages (Fig. 1c) suggesting that miR-130b could be involved in morula and blastocyst formation. [score:3]
To understand the role of miR-130b on oocyte companion cells, 7.5 × 10 [4] cells/ml of granulosa or cumulus cells were seeded and sub-confluent cells were transfected with 50 nM of miR-130b precursor, inhibitor, or mismatched using Lipofectamine 2000 reagent (Invitrogen). [score:3]
However overexpression of miR-130b didn’t affect the amount of cholesterol synthesis by the cumulus cells (Fig. 8b), but significantly reduced the secretion of the cholesterol to the culture media (Fig. 8b). [score:3]
However, significantly higher expression of miR-130b was detected in the morula and blastocyst embryonic stages. [score:3]
The transcript levels of miR-130b (a), SMAD5 (b) and MSK1 (c) in blastocysts derived from zygotes injected with miR-130b precursor, miR-130b inhibitor, scrambled miRNA or uninjected group. [score:3]
zygotes First cleavage 48 hpi (%) Morula rate (%) Blastocyst rate (%) miR-130b precursor 490 77.1 ± 5.132.7 ± 3.8 [a] 28.1 ± 2.2 [a] miR-130b inhibitor 451 76.1 ± 0.923.2 ± 2.5 [b] 21.3 ± 1.9 [b] Scramble miRNA 454 79.9 ± 1.227.3 ± 3.8 [ab] 25.9 ± 3.1 [a] Uninjected control 326 79.4 ± 5.132.7 ± 2.4 [a] 27.2 ± 2.6 [a] Different letters of superscripts in the same column indicate significant difference (P ≤ 0.05) between treatment groups. [score:3]
Twenty-two hours after, the cumulus cells were removed and cumulus-free presumptive zygotes were microinjected with 10 pl/zygote of 50 nM miR-130b precursor, inhibitor, or scrambled sequence. [score:3]
This was also reflected in oocyte companion cells where the inhibition of miR-130b reduced the proliferation rate and the glucose metabolism activity. [score:3]
The opposite phenomenon was observed in MII oocytes derived from miR-130b inhibitor injected oocytes. [score:3]
Sub-confluent cells were then transfected with miR-130b inhibitor, precursor, or scramble sequence. [score:3]
However, the role of miR-130b in bovine granulosa and cumulus cell development, oocyte maturation, and preimplantation embryonic development is not yet known. [score:3]
To determine whether miR-130b is involved in glycolysis, cumulus cells were transfected with 50 nM of miR-130b precursor, inhibitor, or mismatch in 24-wells plate in serum-free medium. [score:3]
The red color indicates the expression level of miR-130b or scramble miRNA probe, while the blue color indicates nuclear staining using 4′,6-diamidino-2-phenylindole (DAPI). [score:3]
The luciferase activity in cumulus cells co -transfected with and miR-130b precursor, miR-130b inhibitor, or scramble sequence with pmirGLO vector construct harboring the 3′ UTRs of EIF2C1, DDX2, EIF2C4, MEOX2 and DOC1R. [score:3]
In addition, EIF2C1, DDX6, DOC1R, and MEOX2 genes were also targeted by miR-130b, although the evidence was not as strong as that of the MSK1 and SMAD5 genes (Additional file 3: Fig. S1). [score:3]
Accordingly, the expression level of miR-130b was not significantly altered from zygote until 8-cell stages. [score:3]
Indeed, in current study, both the qPCR and in situ hybridization indicated that in addition to the oocytes, miR-130b is expressed in bovine oocyte surrounding somatic cells and preimplantation embryos. [score:3]
of oocytes First polar body extrusion (%) miR-130b precursor 35686.6 ± 3.7 [a] miR-130b inhibitor 48972.8 ± 5.9 [b] Scrambled 48883.9 ± 3.5 [a, c] Uninjected 39680.1 ± 4.0 [c] Different letters of superscripts in the same column indicate significant difference (P ≤ 0.05) between treatment groups. [score:3]
Therefore, to determine the role of miR-130b in the energy metabolic activity of the oocyte companion cells, cumulus cells were transfected with miR-130b precursors or inhibitor. [score:3]
In addition, assessment of the nuclear maturation status of the oocytes using Hoechst-33,342 indicated that significantly higher proportion of oocytes were arrested at telephase-1 and significantly lower proportion of oocytes were reached the MII stage in the miR-130b inhibitor injected oocyte group (Table 2). [score:3]
The luciferase assay showed that SMAD5 and MSK1 genes were identified as the direct targets of miR-130b. [score:3]
Fig. 1The expression patterns of miR-130b in (a) mature (MO) and immature (IMO) oocytes, (b) immature (IMCC) cumulus, mature (MCC) cumulus and granulosa (GC) cells, (c) preimplantation stage embryos. [score:3]
The bars graphs represent the mean ± standard deviation (SD) and the data was analyzed from three independent biological samples of MO, IMO IMCC, MCC, zygotes; 2-cell, 4-cell, and 8-cell stage embryos; morula; and blastocysts To understand the role of miR-130b in oocyte companion cells during oocyte maturation, granulosa and cumulus cells were transfected with miR-130b precursor, miR-130b inhibitor, or miRNA scramble sequence. [score:3]
For this, first the expression pattern of miR-130b was analyzed in different preimplantation stage embryos. [score:3]
From that study, including miR-130b, a total of 59 miRNAs were differentially expressed between the two oocyte groups. [score:3]
The total number of live cumulus (a) and granulosa (b) cells 24 and 48 h post miR-130b precursor, miR-130b inhibitor, scramble miRNA transfection or untransfected cell groups. [score:3]
The expression pattern of miR-130b was analyzed in immature and in vitro-matured oocytes, cumulus cells and granulosa cells. [score:3]
The expression patterns of miR-130b in oocytes, oocyte companion cells and preimplantation stage embryos. [score:3]
For this, first MSK1 (RPS6KA5) and SMAD5 were identified as the target genes of miR-130b by integrating in silico and wet-lab analyses. [score:3]
The bars graphs indicate the mean ± standard deviation (SD) from three independent biological samples of MO, IMO IMCC, MCC, zygotes; 2-cell, 4-cell, and 8-cell stage embryos; morula; and blastocysts In addition to expression profiling, in situ localization of miR-130b was performed in pre-antral and antral follicles and preimplantation embryos. [score:3]
Increased expression of miR-130b was also found to be associated with the proliferation of pancreatic cancer [34]. [score:3]
The expression pattern of miR-130b was analyzed in cDNA samples obtained from granulosa cells, immature and matures oocytes and their corresponding cumulus cells, and preimplantation embryos using quantitative real time PCR (qPCR). [score:3]
Bars graphs represent the mean ± standard deviation (SD) and the experiment was repeated three times in three independent samples A similar analysis was performed for the MSK1 gene to confirm if MSK1 is also targeted by miR-130b. [score:3]
To confirm this, gain-and loss-of miR-130b function was performed in cumulus and granulosa cells using its precursor and inhibitor as suggested previously [44]. [score:3]
This may suggest that miR-130b could be embryonic miRNA whereas its target genes are maternal transcripts. [score:3]
Lai KW Koh KX Loh M Tada K Subramaniam MM Lim XY MicroRNA-130b regulates the tumour suppressor RUNX3 in gastric cancerEur J Cancer. [score:3]
Expression analysis of miR-130b in oocyte companion cells, oocytes, and preimplantation embryos using quantitative real time PCR (qPCR). [score:3]
Based on the results obtained using qPCR and in situ localization, it was suggested that miR-130b could be involved in regulating the granulosa and cumulus cell function, oocyte maturation and preimplantation embryo development. [score:3]
Protein analysis of genes targeted by miR-130b was performed in oocyte companion cells, oocytes, and the embryos of different treatment groups using the western blotting technique. [score:3]
Bars graphs represent the mean ± standard deviation (SD) and the experiment was repeated three times in three independent samples A similar analysis was performed for the MSK1 gene to confirm if MSK1 is also targeted by miR-130b. [score:3]
In addition, miR-130b is highly expressed in mouse mammary tumor [27], liver cancer [28, 29], mesenchyma stromal cells [30], fibroblast cells [31], gastric cells [32], human mammary epithelial cells [27], and glioma cells [33]. [score:3]
The assay results indicated that while overexpression of miR-130b increased, inhibition of miR-130b reduced the granulosa and cumulus cell proliferation compared to scramble and untransfected cell groups (Fig. 7c & d). [score:3]
Therefore, SMAD5 and MSK1 were selected as the validated target genes of miR-130b for further functional analysis. [score:3]
The oocytes were then injected with 10 pl of 50 nM miR-130b precursor, miR-130b inhibitor, or mismatch. [score:3]
Therefore, further overexpressing miR-130b may not have a significant effect on oocyte maturation. [score:3]
Validation of genes targeted miR-130b. [score:3]
This may indicate that miR-130b could be involved in cumulus and granulosa cell proliferation by modulating the expression patterns of SMAD5 and MSK1 genes. [score:3]
For this, the MII stage oocytes obtained from GV oocytes injected with miR-130b precursor, inhibitor, or scramble miRNA sequence were subjected for mitochondrial assay using mitochondrion-specific dye. [score:2]
Fig. 10 The mitochondrial activity in MII oocytes derived from GV oocytes injected with miR-130b precursor, miR-130b inhibitor or scrambled miRNA Once we comprehended that miR-130b is involved in oocyte maturation, the role of miR-130b was investigated during the bovine preimplantation embryonic development. [score:2]
MicroRNA-130b targets the SMAD5 and MSK1 genes. [score:2]
Fig. 4Experimental validation of the target genes of the miR-130b using dual luciferase assay. [score:2]
Oocyte Embryo miR-130b Mitochondrial activity Development of oocyte starts in the fetal ovary, while final oocyte growth and maturation occurs during the adulthood [1]. [score:2]
Since miR-130b was found to regulate the cumulus cells survival and proliferation, cumulus cell glucose metabolic activity and cholesterol production; we further sought to understand whether miR-130b is involved in oocyte maturation. [score:2]
Accordingly, the cumulus cells co -transfected with miR-130b precursor and the pmiRGLO vector harboring the MSK1 3′-UTR (MSK1GLO) exhibited a significant decrease in luciferase activity, compared to cells co -transfected with miR-130b precursor and pmiRGLO vector harboring of the mismatch sequence (Non-targetGLO), or cumulus cells co -transfected with scramble miRNA and MSK1GlO, or cells transfected with MSK1GlO alone (Fig. 4b). [score:2]
The genes targeted by miR-130b were identified using dual-luciferase reporter assay. [score:2]
Following this, the miR-130b target genes were in silico analyzed and validated using the dual luciferase assay. [score:2]
Thus, it is speculated that miR-130b could affect the oocyte maturation by regulating the proliferation and metabolic activity of the surrounding cells gene. [score:2]
This study demonstrated that in vitro functional modulation of miR-130b affected granulosa and cumulus cell proliferation and survival, oocyte maturation, morula and blastocyst formation suggesting that miR-130b is involved in bovine oocyte maturation and preimplantation embryo development. [score:2]
In oocyte companion cells, the expression of miR-130b was relatively higher in granulosa cells (GC) compared to the cumulus cells surrounded the immature (IMCC) or cumulus cells surrounded the mature oocytes (MCC) (Fig. 1b). [score:2]
Accordingly, the lactate production was significantly increased in cumulus cells transfected with miR-130b precursor, and reduced in cells transfected with miR-130b inhibitor compared to scramble transfected and untransfected cell groups (Fig. 8a). [score:2]
Accordingly, the signal intensity of miR-130b tended to be stronger in oocyte surrounding cells compared to oocytes in antral follicles, while in preantral follicles, the expression level tended to be similar in oocytes and their surrounding cells (Fig. 2). [score:2]
The mitochondrial assay was performed in oocytes 22 h post miR-130b precursor, inhibitor, or scrambled sequence injection. [score:2]
The phenotypic data showed that the first polar body extrusion in oocytes injected with miR-130b inhibitor was significantly lower and tended to be higher in precursor injected group, compared to the oocytes injected with scramble RNA (Table 1). [score:2]
The proliferation rate of cumulus (c) and granulosa (d) cells post miR-130b precursor, miR-130b inhibitor, scramble miRNA transfection and untransfected cell groups determined using MTT assay. [score:2]
Therefore, here, we aimed to examine the role of miR-130b in bovine granulosa and cumulus cell function, oocyte maturation, and preimplantation embryonic development using miRNA gain- and loss-of-function approaches. [score:2]
List of primers used for validation of the miR-130b target genes. [score:2]
The results showed that the mitochondrial signal intensity was lower and higher in the MII oocytes derived from miR-130b inhibitor and precursor injected GV oocytes, respectively compared to scramble miRNA injected and uninjected control oocyte groups (Fig. 10). [score:2]
Moreover, to investigate the role of miR-130b during preimplantation embryo development, zygote stage embryos were microinjected with miR-130b precursor or inhibitor and the cleavage rate, morula and blastocyst formation was analyzed in embryos derived from each zygote group after in vitro culture. [score:2]
Twenty-four hours after transfection, the expression level of miR-130b was increased in precursor transfected cumulus (Fig. 6a) and granulosa (Fig. 6b) cells compared to scrambled transfected and untransfected cell groups. [score:2]
In this regard, our data demonstrated that modulation of miR-130b during in vitro oocyte maturation significantly affected the mitochondrial activity in the MII oocytes indicating the possible role of miR-130b in regulating the mitochondrial activity during in vitro bovine oocyte maturation. [score:2]
Yu T Cao R Li S Fu M Ren L Chen W MiR-130b plays an oncogenic role by repressing PTEN expression in esophageal squamous cell carcinoma cellsBMC Cancer. [score:2]
The result showed that, 24 and 48 h post transfection, the total live cells were significantly increased in cumulus and granulosa cells transfected with miR-130b precursor and decreased in cells transfected with miR-130b inhibitor compared to the control groups (Fig. 7a & b). [score:2]
The results showed that the cholesterol level was significantly increased in the miR-130b inhibitor transfected compared to scramble transfected and untransfected cell groups. [score:2]
The signal intensity of miR-130b in primordial (a), primary follicle (b), secondary follicle (c) and antral follicles (d). [score:1]
In situ localization of miR-130 in ovarian section was performed according to previously described protocols [17, 36]. [score:1]
In the first phase of the study, the expression profile of miR-130b in granulosa cells, immature oocytes and corresponding cumulus cells, matured oocyte and corresponding cumulus cells and preimplantation embryos was investigated. [score:1]
In situ localization of miR-130b in follicular cells and preimplantation embryos. [score:1]
Fig. 2In situ localization of miR-130b in ovarian sections using 3′-digoxigenin labeled locked nucleic acid (LNA) microRNA probe. [score:1]
From that specific study, miR-130b was among the several miRNAs whose abundance level was altered during in vitro oocyte maturation. [score:1]
In situ localization of miR-130b in ovarian tissue. [score:1]
MicroRNA-130b regulates cholesterol biosynthesis in cumulus cells. [score:1]
MicroRNA-130b regulates glucose metabolism in cumulus cell. [score:1]
Embryos were incubated overnight with 3′-Digoxigenin (DIG) labeled LNA -modified oligonucleotide probes (1 pM) for mir-130b, together with scrambled RNAs (Exiqon, Vedbaek, Denmark) in hybridization buffer in a humidified chamber at the temperature 20 °C below the T m of probes. [score:1]
Of those, miR-130b was more interesting, as it belongs to the miR-130 family and this miRNA is known to be conserved in vertebrates [12]. [score:1]
The threshold cycle (C [t]) values of miR-130b and the endogenous controls were recorded using Sequence Detection Software (SDS v1.2.1, Applied Biosystems, USA). [score:1]
MicroRNA-130b regulates mitochondrial activity in oocytes during in vitro maturation. [score:1]
The samples were then incubated with 3′-Digoxigenin (DIG) labeled LNA -modified oligonucleotide probes (1 pM) of miR-130b, U6 or scramble RNAs (Exiqon, Vedbaek, Denmark) in hybridization buffer in a humidified chamber at the temperature 20 °C below the T m of probes. [score:1]
Whole mount in situ localization of miR-130b in oocytes and preimplantation embryo. [score:1]
The analysis was performed from oocytes obtained from three independent in vitro maturation runs Once we realized that inhibition of miR-130b was significantly reduced the proportion of oocytes that reached the MII stage, the effect of miR-130b on the quality of MII oocytes was investigated by determining the mitochondrial activity. [score:1]
The result showed that the luciferase activity was significantly reduced in the cumulus cells co -transfected with miR-130b precursor and the pmiRGLO vector harboring of the SMAD5 3′-UTR (SMAD5GlO). [score:1]
Apart from these, modulation of miR-130b altered the lactate production and cholesterol biosynthesis in cumulus cells. [score:1]
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[+] score: 15
The expression of miR-223, miR-184, miR-132, miR-1246 and miR-130b were up-regulated while miR-196a, miR-205, miR-200b, miR-31 and miR-145 were down-regulated, which was in agreement with the high-throughput sequencing results (Figure 4). [score:9]
miRNA Target Genes miR-1246 ATP2B4, MAP3K1, ADCK3, PSD2, SLC5A1 miR-130b EXOC3L1, TIE1, BAZ2B, C3, GRAMD1C miR-145 HSD3B7, SLCO4A1, PDIA4, ACADL, PTPN11, KRT9, RASSF6, RNF43, LAMC2 miR-196a ADAP1, GPR97, POMT1 miR-200b ARID3A, MLXIP, GPR110 miR-205 IL13RA2, COL5A2, ADM, CXCR2, XPO6, SPSB1, FMO5, PSMF1 miR-31 MEX3D, PFKFB3, ST3GAL3, IL2RB, ANKRD32, MGST1 miR-184 HSPA1L, SLC25A15, HEG1, MAPRE2, ACP6, SYNE2 miR-223 TMEM165 miR-132 IQCA1 Figure 5 Gene ontology statistics. [score:3]
miRNA Target Genes miR-1246 ATP2B4, MAP3K1, ADCK3, PSD2, SLC5A1 miR-130b EXOC3L1, TIE1, BAZ2B, C3, GRAMD1C miR-145 HSD3B7, SLCO4A1, PDIA4, ACADL, PTPN11, KRT9, RASSF6, RNF43, LAMC2 miR-196a ADAP1, GPR97, POMT1 miR-200b ARID3A, MLXIP, GPR110 miR-205 IL13RA2, COL5A2, ADM, CXCR2, XPO6, SPSB1, FMO5, PSMF1 miR-31 MEX3D, PFKFB3, ST3GAL3, IL2RB, ANKRD32, MGST1 miR-184 HSPA1L, SLC25A15, HEG1, MAPRE2, ACP6, SYNE2 miR-223 TMEM165 miR-132 IQCA1 Figure 5 Gene ontology statistics. [score:3]
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[+] score: 11
A recent study reported abundant expression of MIR130b in goat mammary gland tissue and revealed a negative adipogenic effect potentially through inhibition of PPARGC1A [46]. [score:5]
Similarly to what happens for PPARG, MIR130b potently repressed PPARGC1A expression by targeting both the PPARGC1A mRNA coding region and 3’UTR [46]. [score:5]
It might be possible that MIR130A does not function in the same manner as MIR130B. [score:1]
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[+] score: 7
However, few miRNAs had their targets identified, such as miR-130 that regulates PPARγ and inhibits adipogenesis [23], and miR-181a that regulates TNFα and increases adipogenesis [24]. [score:7]
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[+] score: 6
When primary cultured porcine adipocytes are exposed to miR-130b-enriched micro-vesicles, peroxisome proliferator-activated receptor γ (PPARG) expression is downregulated [19]. [score:6]
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[+] score: 5
Other miRNAs from this paper: bta-mir-26a-2, bta-mir-29a, bta-let-7f-2, bta-mir-16b, bta-mir-21, bta-mir-221, bta-mir-222, bta-mir-30d, bta-mir-99a, bta-mir-145, bta-mir-181a-2, bta-mir-199a-1, bta-mir-27b, bta-mir-142, bta-mir-181b-2, bta-mir-30e, bta-mir-92a-2, bta-let-7d, bta-mir-132, bta-mir-181c, bta-mir-191, bta-mir-199b, bta-mir-214, bta-mir-29b-2, bta-mir-29c, bta-mir-455, bta-let-7g, bta-mir-10b, bta-mir-24-2, bta-let-7a-1, bta-mir-150, bta-let-7f-1, bta-mir-122, bta-let-7i, bta-mir-34c, bta-mir-363, bta-let-7a-2, bta-let-7a-3, bta-let-7b, bta-let-7c, bta-let-7e, bta-mir-195, bta-mir-34a, bta-mir-365-1, bta-mir-99b, bta-mir-100, bta-mir-129-1, bta-mir-129-2, bta-mir-130a, bta-mir-133a-2, bta-mir-133a-1, bta-mir-143, bta-mir-146b, bta-mir-146a, bta-mir-155, bta-mir-181d, bta-mir-182, bta-mir-183, bta-mir-184, bta-mir-24-1, bta-mir-196a-2, bta-mir-196a-1, bta-mir-199a-2, bta-mir-212, bta-mir-26a-1, bta-mir-28, bta-mir-29d, bta-mir-32, bta-mir-335, bta-mir-338, bta-mir-339a, bta-mir-346, bta-mir-365-2, bta-mir-378-1, bta-mir-383, bta-mir-409a, bta-mir-449a, bta-mir-449b, bta-mir-449c, bta-mir-592, bta-mir-708, bta-mir-92a-1, bta-mir-92b, bta-mir-29e, bta-mir-29b-1, bta-mir-1271, bta-mir-1249, bta-mir-181a-1, bta-mir-181b-1, bta-mir-2285a, bta-mir-2285d, bta-mir-2285b-1, bta-mir-2332, bta-mir-199c, bta-mir-2389, bta-mir-2285c, bta-mir-2404-1, bta-mir-449d, bta-mir-2411, bta-mir-2446, bta-mir-339b, bta-mir-2404-2, bta-mir-2483, bta-mir-424, bta-mir-378-2, bta-mir-409b, bta-mir-2285e-1, bta-mir-2285e-2, bta-mir-2285f-1, bta-mir-2285f-2, bta-mir-2285g-1, bta-mir-2285h, bta-mir-2285i, bta-mir-2285j-1, bta-mir-2285j-2, bta-mir-2285k-1, bta-mir-2285l, bta-mir-2285o-1, bta-mir-2285o-2, bta-mir-2285n-1, bta-mir-2285n-2, bta-mir-2285p, bta-mir-2285m-1, bta-mir-2285m-2, bta-mir-378b, bta-mir-2285n-3, bta-mir-2285n-4, bta-mir-2285o-3, bta-mir-2285o-4, bta-mir-2285m-3, bta-mir-378c, bta-mir-2285m-4, bta-mir-2285o-5, bta-mir-2285m-5, bta-mir-2285n-5, bta-mir-2285n-6, bta-mir-2285n-7, bta-mir-2285k-2, bta-mir-2285k-3, bta-mir-2285k-4, bta-mir-2285k-5, bta-mir-2285q, bta-mir-2285r, bta-mir-2285s, bta-mir-2285t, bta-mir-2285b-2, bta-mir-2285v, bta-mir-2285g-2, bta-mir-2285g-3, bta-mir-2285af-1, bta-mir-2285af-2, bta-mir-2285y, bta-mir-2285w, bta-mir-2285x, bta-mir-2285z, bta-mir-2285u, bta-mir-2285aa, bta-mir-2285ab, bta-mir-2285ac, bta-mir-2285ad, bta-mir-2285ae, bta-mir-378d, bta-mir-2285ag, bta-mir-2285ah, bta-mir-2285ai, bta-mir-2285aj, bta-mir-2285ak, bta-mir-2285al, bta-mir-2285am, bta-mir-2285ar, bta-mir-2285as-1, bta-mir-2285as-2, bta-mir-2285as-3, bta-mir-2285at-1, bta-mir-2285at-2, bta-mir-2285at-3, bta-mir-2285at-4, bta-mir-2285au, bta-mir-2285av, bta-mir-2285aw, bta-mir-2285ax-1, bta-mir-2285ax-2, bta-mir-2285ax-3, bta-mir-2285ay, bta-mir-2285az, bta-mir-2285an, bta-mir-2285ao-1, bta-mir-2285ao-2, bta-mir-2285ap, bta-mir-2285ao-3, bta-mir-2285aq-1, bta-mir-2285aq-2, bta-mir-2285ba-1, bta-mir-2285ba-2, bta-mir-2285bb, bta-mir-2285bc, bta-mir-2285bd, bta-mir-2285be, bta-mir-2285bf-1, bta-mir-2285bf-2, bta-mir-2285bf-3, bta-mir-2285bg, bta-mir-2285bh, bta-mir-2285bi-1, bta-mir-2285bi-2, bta-mir-2285bj-1, bta-mir-2285bj-2, bta-mir-2285bk, bta-mir-2285bl, bta-mir-2285bm, bta-mir-2285bn, bta-mir-2285bo, bta-mir-2285bp, bta-mir-2285bq, bta-mir-2285br, bta-mir-2285bs, bta-mir-2285bt, bta-mir-2285bu-1, bta-mir-2285bu-2, bta-mir-2285bv, bta-mir-2285bw, bta-mir-2285bx, bta-mir-2285by, bta-mir-2285bz, bta-mir-2285ca, bta-mir-2285cb, bta-mir-2285cc, bta-mir-2285cd, bta-mir-2285ce, bta-mir-2285cf, bta-mir-2285cg, bta-mir-2285ch, bta-mir-2285ci, bta-mir-2285cj, bta-mir-2285ck, bta-mir-2285cl, bta-mir-2285cm, bta-mir-2285cn, bta-mir-2285co, bta-mir-2285cp, bta-mir-2285cq, bta-mir-2285cr-1, bta-mir-2285cr-2, bta-mir-2285cs, bta-mir-2285ct, bta-mir-2285cu, bta-mir-2285cv-1, bta-mir-2285cv-2, bta-mir-2285cw-1, bta-mir-2285cw-2, bta-mir-2285cx, bta-mir-2285cy, bta-mir-2285cz, bta-mir-2285da, bta-mir-2285db, bta-mir-2285dc, bta-mir-2285dd, bta-mir-2285de, bta-mir-2285df, bta-mir-2285dg, bta-mir-2285dh, bta-mir-2285di, bta-mir-2285dj, bta-mir-2285dk, bta-mir-2285dl-1, bta-mir-2285dl-2, bta-mir-2285dm
In addition, the expression of 12 miRNA families including miR-130 (a, b), bta-miR-181 (a, b, c, d), bta-miR-199 (a-3p, a-5p, b, c), bta-miR-2285 (k, t), bta-miR-2411 (- 3p,-5p), bta-miR-2483 (- 3p,-5p), bta-miR-29 (a, b), bta-miR-339 (a, b), bta-miR-365 (- 3p,-5p), bta-miR-455 (- 3p,-5p), bta-miR-92, and bta-miR-99 (a,-5p, b) were differentially expressed between the granulosa cells of SF and DF (Table 1). [score:5]
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[+] score: 5
Other miRNAs from this paper: hsa-let-7a-1, hsa-let-7a-2, hsa-let-7a-3, hsa-let-7b, hsa-let-7c, hsa-let-7d, hsa-let-7e, hsa-let-7f-1, hsa-let-7f-2, hsa-mir-15a, hsa-mir-17, hsa-mir-18a, hsa-mir-19a, hsa-mir-19b-1, hsa-mir-19b-2, hsa-mir-20a, hsa-mir-22, hsa-mir-24-1, hsa-mir-24-2, hsa-mir-25, hsa-mir-27a, hsa-mir-29a, hsa-mir-30a, hsa-mir-92a-1, hsa-mir-92a-2, hsa-mir-98, hsa-mir-99a, hsa-mir-29b-1, hsa-mir-29b-2, hsa-mir-106a, hsa-mir-148a, hsa-mir-30c-2, hsa-mir-30d, hsa-mir-10a, hsa-mir-10b, hsa-mir-181a-2, hsa-mir-181b-1, hsa-mir-181c, hsa-mir-182, hsa-mir-181a-1, hsa-mir-221, hsa-let-7g, hsa-let-7i, hsa-mir-1-2, hsa-mir-15b, hsa-mir-27b, hsa-mir-30b, hsa-mir-130a, hsa-mir-152, hsa-mir-191, hsa-mir-9-1, hsa-mir-9-2, hsa-mir-9-3, hsa-mir-185, hsa-mir-193a, hsa-mir-320a, hsa-mir-200c, hsa-mir-1-1, hsa-mir-181b-2, hsa-mir-29c, hsa-mir-30c-1, hsa-mir-99b, hsa-mir-130b, hsa-mir-30e, hsa-mir-363, hsa-mir-374a, hsa-mir-375, hsa-mir-378a, hsa-mir-148b, hsa-mir-331, hsa-mir-339, hsa-mir-423, hsa-mir-20b, hsa-mir-491, hsa-mir-193b, hsa-mir-181d, hsa-mir-92b, hsa-mir-320b-1, hsa-mir-320c-1, hsa-mir-320b-2, hsa-mir-378d-2, bta-mir-29a, bta-let-7f-2, bta-mir-148a, bta-mir-18a, bta-mir-20a, bta-mir-221, bta-mir-27a, bta-mir-30d, bta-mir-320a-2, bta-mir-99a, bta-mir-181a-2, bta-mir-27b, bta-mir-30b, bta-mir-106a, bta-mir-10a, bta-mir-15b, bta-mir-181b-2, bta-mir-193a, bta-mir-20b, bta-mir-30e, bta-mir-92a-2, bta-mir-98, bta-let-7d, bta-mir-148b, bta-mir-17, bta-mir-181c, bta-mir-191, bta-mir-200c, bta-mir-22, bta-mir-29b-2, bta-mir-29c, bta-mir-423, bta-let-7g, bta-mir-10b, bta-mir-24-2, bta-mir-30a, bta-let-7a-1, bta-let-7f-1, bta-mir-30c, bta-let-7i, bta-mir-25, bta-mir-363, bta-let-7a-2, bta-let-7a-3, bta-let-7b, bta-let-7c, bta-let-7e, bta-mir-15a, bta-mir-19a, bta-mir-19b, bta-mir-331, bta-mir-374a, bta-mir-99b, hsa-mir-374b, hsa-mir-320d-1, hsa-mir-320c-2, hsa-mir-320d-2, bta-mir-1-2, bta-mir-1-1, bta-mir-130a, bta-mir-152, bta-mir-181d, bta-mir-182, bta-mir-185, bta-mir-24-1, bta-mir-193b, bta-mir-29d, bta-mir-30f, bta-mir-339a, bta-mir-374b, bta-mir-375, bta-mir-378-1, bta-mir-491, bta-mir-92a-1, bta-mir-92b, bta-mir-9-1, bta-mir-9-2, bta-mir-29e, bta-mir-29b-1, bta-mir-181a-1, bta-mir-181b-1, bta-mir-320b, bta-mir-339b, bta-mir-19b-2, bta-mir-320a-1, bta-mir-193a-2, bta-mir-378-2, hsa-mir-378b, hsa-mir-320e, hsa-mir-378c, bta-mir-148c, hsa-mir-374c, hsa-mir-378d-1, hsa-mir-378e, hsa-mir-378f, hsa-mir-378g, hsa-mir-378h, hsa-mir-378i, hsa-mir-378j, bta-mir-378b, bta-mir-378c, bta-mir-378d, bta-mir-374c, bta-mir-148d
In addition, many miRNA families showed low expression (count number <100) in milk exosomes, such as the miR-1, miR-130, miR-17, miR-10, miR-29, miR-374, mir-9, miR-15 and miR-491 families (Figure 12F), which are routinely expressed in specific tissues [53– 56]. [score:5]
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[+] score: 4
For example, miR-130b, a dairy efficiency negatively associated miRNA with lower expression under AL comparing to both RS and CS in the rumen (Table S7), has been reported to regulate cellular proliferation in muscle. [score:4]
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[+] score: 3
Overall, the transportable exogenous miRNAs predicted in this study are involved in many major biological processes including development, differentiation, cell proliferation, and metabolism [56], e. g. miR-27b, miR-34a, miR-106b, and miR-130 that are related to immune or development [6– 8]. [score:3]
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[+] score: 2
Other miRNAs from this paper: bta-mir-301b, bta-mir-3596
It should be noted that 5 intronic SNPs within NPAS2 gene associated with C13:0. Similarly, two SNPs in each of GRB10, CSGALNACT1, XPNPEP1, MAD1L1 and CLSTN2 genes attained genome wide significance with C13:0. Furthermore, two SNPs within microRNA genes, (rs440208182 on MIR130B/MIR301B and rs480300366 on MIR3596/MIRLET7B) associated with C13:0. Tricosanoic acid (C23:0) and lignoceric acid (C24:0) long chain saturated fatty acids (SFAs) associated significantly with several SNP markers while no associations were recorded for C20:0 and C22:0 (Table S7f,g). [score:1]
It should be noted that 5 intronic SNPs within NPAS2 gene associated with C13:0. Similarly, two SNPs in each of GRB10, CSGALNACT1, XPNPEP1, MAD1L1 and CLSTN2 genes attained genome wide significance with C13:0. Furthermore, two SNPs within microRNA genes, (rs440208182 on MIR130B/MIR301B and rs480300366 on MIR3596/MIRLET7B) associated with C13:0. Tricosanoic acid (C23:0) and lignoceric acid (C24:0) long chain saturated fatty acids (SFAs) associated significantly with several SNP markers while no associations were recorded for C20:0 and C22:0 (Table S7f,g). [score:1]
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This extends the work of Mondou et al. [27] who demonstrated transcription -dependent increases of specific miRNA precursors (miR-21 and miR-130) during oocyte maturation and early embryogenesis. [score:1]
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