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9 publications mentioning rno-mir-376a

Open access articles that are associated with the species Rattus norvegicus and mention the gene name mir-376a. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

1
[+] score: 119
Therefore, the post-transcriptional control of GRP78 by rno-miR-376a has a significant impact on the regulation of GRP78 protein expression and not gene expression, which is evident by the lack of GRP78 transcriptional regulation. [score:7]
To identify the rno-miR-376a -binding site at the 3′-end of GRP78 mRNA, we generated a direct-match miRNA target site and cloned the insert into the multiple cloning site in the luciferase reporter vector from the pMIR-REPORT miRNA Expression Reporter Vector System (Applied Biosystems). [score:6]
Therefore, we measured the expression of GRP78 mRNA, rno-miR-144, rno-miR-376a, and rno-miR-451 using real-time RT-PCR to examine their expression patterns in the context of LHR downregulation (Fig. 1). [score:6]
Effects of Pre-miR-376a (precursor) and Anti-miR-376a (inhibitor) transfection on rat GRP78 mRNA expression in primary rat granulosa cells. [score:5]
In a similar manner, rno-miR-144 and rno-miR-376a expression peaked 12 h after the hCG treatment, and rno-miR-451 expression peaked 24 h after the hCG treatment. [score:5]
The results presented in this study (Fig. 4 and Fig. 5) demonstrate that rno-miR-376a decreases GRP78 protein production by translational repression without altering GRP78 mRNA levels and that the transfection of rno-miR-376a into granulosa cells repressed protein expression by approximately three-fold (Fig. 5). [score:5]
Thus, we believe that rno-miR-376a is involved in the regulation of GRP78 expression in the ovary. [score:4]
Combined with the MicroCosm analysis of miRNA targets, which revealed that 44 miRNAs can bind to the 3′-UTR of GRP78 mRNA, we speculate that multiple miRNAs may constitute a network that is involved in the regulation of GRP78 mRNA, whereas rno-miR-376a is solely identified to be evoked by the ovulatory signal. [score:4]
uk/) revealed that rno-miR-144, rno-miR-376a, and rno-miR-451 can bind to the 3′-UTR of GRP78 mRNA (from bp 2439–2459) and negatively regulate GRP78 expression. [score:4]
Rat GRP78 mRNA, rno-miR-376a expression in primary rat granulosa cells induced by FSH. [score:3]
From these, we narrowed the focus to rno-miR-376a based on the results of the in vitro experiments (Fig. 2) since rno-miR-144 and rno-miR-451 was not induced expression by hCG treatment. [score:3]
Rat GRP78 mRNA, rno-miR-144, rno-miR-376a, and rno-miR-451 expression in primary rat granulosa cells induced by FSH and hCG. [score:3]
The following oligonucleotides were used in this studies: (1) miR-376a, 5′-aatgcactagtACGAGGATTTTCCTCTACGATaagcttaatgc-3′ and 5′-gcattaagcttATCGTAGAGGAAAATCCTCGTactagtgcatt-3′, and (2) rno-miR-376a -binding site sequence at the 3′-end of GRP78 mRNA, 5′-aatgcactagtATGGTAGAAAAAAGTTCCTACaagcttaatgc-3′ and 5′-gcattaagcttGTAGGAACTTTTTTCTACCATactagtgcatt-3′ We transfected HEK293 cells with 200 ng of each vector (pMIR-REPORT luciferase vectors, as described above, and the pMIR-REPORT βgal vector as a control for transfection normalization) and 50 nM precursor or inhibitor using Lipofectamine 2000 Transfection Reagent according to the manufacturer's protocol. [score:3]
The array data along with the bioinformatic analysis provided by MicroCosm Targets, which indicated that several miRNAs bind to the GRP78 mRNA 3′-UTR, led us to focus on rno-miR-144, rno-miR-376a, and rno-miR-451. [score:3]
Time course of rat GRP78 mRNA, rno-miR-144, rno-miR-376a, and rno-miR-451 expression in rat ovaries induced by PMSG and hCG. [score:3]
Moreover, the state of the complementarity between target mRNA and miRNA has been suggested to affect mRNA degradation [34], which is applicable to rno-miR-376a because the introduction of the complete identical sequence of rno-miR-376a into a reporter vector abolished the luciferase activity, whereas the luciferase activity was reduced to 50% using the reporter vector containing the 3′-UTR rno-miR-376a binding site of GRP78 mRNA (Fig. 6). [score:3]
These data demonstrate that the translation of GRP78 was blocked by precursor via a binding site that was the complementary sequence to rno-miR-376a. [score:3]
miR-376a does not affect GRP78 mRNA expression. [score:3]
The cells were incubated with FSH (30 ng/mL) and estradiol (10 nM) for 48 h. Pre-miR-376a (precursor) or Anti-miR-376a (inhibitor) was transfected into the cells, and 30 ng/mL hCG was added 12 h later. [score:3]
In conclusion, the present results demonstrate that the induction of rno-miR-376a production by hCG leads to repression of GRP78 translation. [score:3]
Twenty-four hours after seeding, the cells were incubated with FSH (30 ng/mL) and estradiol (10 nM) for 48 h. The cells were then transfected with Pre-miR-376a (precursor) or Anti-miR-376a (inhibitor) (50 nM each) purchased from Ambion (product IDs: PM10504 and AM10504, respectively) using siPORT NeoFX Transfection Agent according to the manufacturer's protocol. [score:3]
HEK293 cells were prepared, and the cells were transfected with 200 ng of each reporter vector with 50 nM Pre-miR-376a (precursor) or Anti-miR-376a (inhibitor) as described in the. [score:3]
We then examined the effects of rno-miR-376a on GRP78 mRNA levels following the transfection of granulosa cells with either precursor or inhibitor (Fig. 4). [score:3]
The following oligonucleotides were used in this studies: (1) miR-376a, 5′-aatgcactagtACGAGGATTTTCCTCTACGATaagcttaatgc-3′ and 5′-gcattaagcttATCGTAGAGGAAAATCCTCGTactagtgcatt-3′, and (2) rno-miR-376a -binding site sequence at the 3′-end of GRP78 mRNA, 5′-aatgcactagtATGGTAGAAAAAAGTTCCTACaagcttaatgc-3′ and 5′-gcattaagcttGTAGGAACTTTTTTCTACCATactagtgcatt-3′ We transfected HEK293 cells with 200 ng of each vector (pMIR-REPORT luciferase vectors, as described above, and the pMIR-REPORT βgal vector as a control for transfection normalization) and 50 nM precursor or inhibitor using Lipofectamine 2000 Transfection Reagent according to the manufacturer's protocol. [score:3]
miR-376a represses GRP78 translation. [score:3]
GRP78 mRNA and rno-miR-376a expression increased significantly 12 h after the addition of hCG into the culture medium and subsequently decreased, which was consistent with in vivo study (Fig. 1). [score:3]
Transfection with Pre-miR-376a miRNA (precursor) and Anti-miR-376a miRNA (inhibitor). [score:3]
Cells were then incubated with FSH (30 ng/mL) and estradiol (10 nM) for 48 h in the same way as described in Fig. 2. The time after 48 h of incubation with FSH and estradiol was considered “0 h. ” Total RNA was isolated, and rno-miR-376a expression levels was determined using real-time RT-PCR at the indicated time. [score:2]
Total RNA was isolated, and GRP78 mRNA (A), rno-miR-144 (B), rno-miR-376a (C), and rno-miR-451 (D) expression levels were measured using real-time RT-PCR as described in the. [score:1]
miR-376a binds to predicted site of GRP78 3′-UTR. [score:1]
Effects of rno-miR-376a on GRP78 protein in granulosa cells. [score:1]
To identify the rno-miR-376a -binding site in the 3′-UTR of GRP78 mRNA, luciferase reporter vectors were generated as described in the. [score:1]
To confirm the presence of an rno-miR-376a binding site on GRP78 mRNA, we constructed a reporter vector that contained a putative rno-miR-376a binding site sequence (bp 2439–2459) in the 3′-UTR downstream of a Renilla luciferase coding region. [score:1]
One can argue whether this reduction of GRP78 protein by rno-miR-376a has significant meaning for physiological functions in the ovary. [score:1]
The amount of rat GRP78 mRNA, rno-miR-144, rno-miR-376a, and rno-miR-451 in the hCG 0 h group was set at 1. Data were normalized to 18S rRNA (for GRP78 mRNA) and 4.5S RNA(H) (for rno-miR-144, rno-miR-376a, and rno-miR451) levels in each sample and represent the mean ±SE of three independent experiments. [score:1]
This result indicates that rno-miR-376a binds to the 3′-end of GRP78 mRNA from bp 2439–2459 in the 3′-UTR. [score:1]
The luciferase activity in the cells transfected with the reporter vector containing the putative rno-miR-376a binding site was reduced by approximately 50% 24 h after the cells were transfected with precursor. [score:1]
The amounts of rno-miR-376a in the 0 h group were set at 1. Data were normalized for 4.5S RNA(H) levels in each sample and represent the mean ±SE of 3 independent experiments. [score:1]
0108997.g006 Figure 6s for the identification of the rno-miR-376a -binding site in the 3′-UTR of GRP78 mRNA. [score:1]
To verify the effect of the miRNA transfection, we generated a reporter vector that included the whole miR-376a sequence. [score:1]
To avoid the influence of endogenous rno-miR-376a, we used HEK293 cells rather than rat granulosa cells (Fig. 6). [score:1]
Next, we investigated the effects of rno-miR-144, rno-miR-376a, and rno-miR-451 on GRP78 mRNA expression in granulosa cells isolated from DES -treated immature rats (Fig. 2). [score:1]
Rat GRP78 mRNA (A), rno-miR-144 (B), rno-miR-376a (C), and rno-miR-451 (D) expression levels were measured using real-time RT-PCR as described in the. [score:1]
The amounts of GRP78 mRNA, rno-miR-144, rno-miR-376a, and rno-miR-451 in the hCG 0 h group were set at 1. Data were normalized for 18S rRNA (for GRP78 mRNA) and 4.5S RNA(H) (for rno-miR-144, rno-miR-376a, and rno-miR-451) levels in each sample and represent the mean ±SE of 3 independent experiments. [score:1]
Furthermore, miR-376a did not show a substantial change in the absence of hCG treatment (Fig. 3), confirming that hCG has an important role for the induction of miR-376a. [score:1]
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2
[+] score: 23
0018613.g004 Figure 4Three general trends of miRNA expression trajectories were observed for Wistar islets at 2.8G vs 16.7G: i) increased expression as exhibited by rno-miR-132, rno-miR-212 and rno-miR-409-3p, ii) decreased expression as in the case of rno-miR-124, rno-miR-142-3p, rno-miR-375, rno-miR-335, rno-miR-130a and rno-miR-708 and, iii) no change as seen in rno-miR-376a, rno-miR-142-5p and rno-miR-433. [score:7]
Three general trends of miRNA expression trajectories were observed for Wistar islets at 2.8G vs 16.7G: i) increased expression as exhibited by rno-miR-132, rno-miR-212 and rno-miR-409-3p, ii) decreased expression as in the case of rno-miR-124, rno-miR-142-3p, rno-miR-375, rno-miR-335, rno-miR-130a and rno-miR-708 and, iii) no change as seen in rno-miR-376a, rno-miR-142-5p and rno-miR-433. [score:7]
Using assays [24] we validated the expression of ten most upregulated miRNAs from the significant list, and also prioritizing miRNAs which appeared in previous studies on pancreatic islets or insulin-secreting beta cell lines such as miR-124, miR-376a, miR-132 and miR-212. [score:5]
In general, aside from more significant changes in expression levels of miRNAs at 24 h incubation compared to 1 h incubation, three trends in terms of expression changes are also observed in the Wistar islet upon stimulation at 16.7G as compared to 2.8G: i) increasing miRNA levels, as displayed by rno-miR-132, rno-miR-212 and rno-miR-409-3p, ii) decreasing miRNA levels as exhibited by rno-miR-124, rno-miR-142-3p, rno-miR-375, rno-miR-335, rno-miR-130a and rno-miR-708, and iii) no significant change as displayed by rno-miR-376a, rno-miR-142-5p and rno-miR-433. [score:3]
However, despite these attempts in the GK islet to attain normal levels of miRNAs, failures were seen in most GK miRNAs wherein the levels at 16.7G either overshoot those of Wistar's as in rno-miR-142-3p, rno-miR-142-5p, rno-miR-375 and rno-miR-124, or completely miss the normal levels as in the case of rno-miR-335 and rno-miR-376a (Fig. 4). [score:1]
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3
[+] score: 14
Importantly, in the case of mir-376a-5p, mir-376b-5p, mir-376a-3p and mir-376-3p the edited nucleoside resides in the seed region, which is instrumental in targeting the miRNA to its target and could therefore lead to an altered target spectrum. [score:7]
The most abundantly edited miRNA is mir-376b, in the brain (Figure 2B), fitting with the observation that in murine and human brains, mir-376a/b/c are targets of the RNA editing machinery [51, 52]. [score:3]
Furthermore, we confirmed specific A to I editing in brain for mir-376a/b/c and identified mir-377 as a novel editing target. [score:3]
We find the mir-376a/c isoforms to be edited as well, albeit to a lesser extent (Figure 2B). [score:1]
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4
[+] score: 14
Cluster III included miR-141 and miR-376a and had increased expression at P0 and then a decreased expression at P2 compared to E20. [score:4]
MiR-125b-5p [38] as well as miR-23a and miR-376a are up-regulated in pancreatic cancer [32], [39]. [score:4]
The exeptions were miR-376a, which show a similar expression pattern as miR-376b-3p; and miR-23a, whose levels did not significantly rise from E20 to P2 (Table S1). [score:3]
Even though miR-376a, -376b-3p and -451 were not detected in INS-1E cells, they were detected in some islet cells at E20 and P0. [score:1]
Although ISH is not quantitative this method confirmed the northern blots, since we generally detected a lower signal of miR-376a at P0 and P2 than at E20. [score:1]
MiR-376a, -376b-3p and -451 were localized to exocrine cells (miR-451 only at E20). [score:1]
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5
[+] score: 11
b Heat map of miRNA expression profile in normal mammary tissue of 50-day-old female offspring (n = 3 per group) Since miR-1897-5p, miR-219-1-3p, and miR-376a# can directly or indirectly modulate several targets (shown in 1: Table S1), we decided to perform Western blot analysis of the following proteins linked to breast cancer: CCAAT/enhancer -binding protein beta (Cebpβ), caspase 3 (Casp3), insulin-like growth factor 1 receptor (Igf1r), protein kinase D1 (Pkd1), and transforming growth factor, beta receptor I (Tgfβr1). [score:7]
There were three miRNAs that were downregulated in both the sperm and the mammary glands of the corn oil-fed fathers and their daughters, respectively: miR-1897-5p, miR-219-1-3p, and miR-376a#. [score:4]
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6
[+] score: 10
Interestingly, the relative abundance of “A” to “I” editing of rno-miR-376 gradually increased during development and surpassed that of wild type isoform at P7, indicating that RNA editing may be a new strategy for the regulation of gene expression during brain development. [score:6]
We found that the predicted target genes of the wild type rno-miR-376 and the “A” to “I” edited isoform are of totally different functional groups (Figure 4F). [score:3]
As a distinguished representative of miRNA editing, rno-miRNA-376 family have been extensively studied [5]. [score:1]
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7
[+] score: 4
Among the miRNAs, miR-34a, miR-32, miR-376a, miR-384-3p, miR-29b and miR-142-3p were the highly overexpressed, with fold induction of 11.8, 24.2, 12.7, 14.4, 11.6 and 19.1, respectively. [score:3]
In the 16 week colonic biopsies, we observed that while all miRNAs trended to increase (versus age-matched saline treated animals) although only 7 miRNAs (miR-34a, miR-21, miR-18, miR-376a, miR-19a, miR-9 and miR-29b) achieved statistical significance (fold inductions of 1.73, 2.72, 2.15, 2.26, 2.18, 1.53, and 1.71,respectively) (Table 2). [score:1]
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8
[+] score: 2
Twelve miRNAs (miR-135a, miR-190, miR-22, miR-347, miR-376*, miR-380*, miR-382, miR-383, miR-702-3p, miR-708, miR-873, and miR-99b*) were regulated only in CCE rats (p < 0.05 vs. [score:2]
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9
[+] score: 2
A similar situation can be found in the human cluster {hsa-mir-368, HP-37, HN-7, hsa-mir-376a} which corresponds to the mouse {mmu-mir-376a, mmu-mir-376b, MP-38}. [score:1]
The fine-grained structure of these loci has some species-specific aspects, as illustrated by Figures 2 and 3. The figures show all the validated miRNAs in these regions, including those with suboptimal prediction scores from the Additional files 7, 8, and 9. We find that some miRNAs that are related in sequence, and have presumably arisen by duplication (such as the mir-368/mir-376-related sequences) have different numbers of copies in rodents and human. [score:1]
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