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Expression and preliminary functional profiling of the let-7 family during porcine ovary follicle atresia.

Cao R, Wu WJ, Zhou XL, Xiao P, Wang Y, Liu HL - Mol. Cells (2015)

Bottom Line: In the current study, real-time quantitative polymerase chain reaction showed that the expression levels of let-7 family members in follicles and granulosa cells were similar to our microarray data, in which miRNAs let-7a, let-7b, let-7c, and let-7i were significantly decreased in early atretic and progressively atretic porcine ovary follicles compared with healthy follicles, while let-7g was highly expressed during follicle atresia.In addition, let-7 target genes were predicted and annotated by TargetScan, PicTar, gene ontology and Kyoto encyclopedia of genes and genomes pathways.Our data provide new insight into the association between the let-7 miRNA family in granulosa cell programmed death.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Most follicles in the mammalian ovary undergo atresia. Granulosa cell apoptosis is a hallmark of follicle atresia. Our previous study using a microRNA (miRNA) microarray showed that the let-7 microRNA family was differentially expressed during follicular atresia. However, whether the let-7 miRNA family members are related to porcine (Sus scrofa) ovary follicular apoptosis is unclear. In the current study, real-time quantitative polymerase chain reaction showed that the expression levels of let-7 family members in follicles and granulosa cells were similar to our microarray data, in which miRNAs let-7a, let-7b, let-7c, and let-7i were significantly decreased in early atretic and progressively atretic porcine ovary follicles compared with healthy follicles, while let-7g was highly expressed during follicle atresia. Furthermore, flow cytometric analysis and Hoechst33342 staining demonstrated that let-7g increased the apoptotic rate of cultured granulosa cells. In addition, let-7 target genes were predicted and annotated by TargetScan, PicTar, gene ontology and Kyoto encyclopedia of genes and genomes pathways. Our data provide new insight into the association between the let-7 miRNA family in granulosa cell programmed death.

No MeSH data available.


Related in: MedlinePlus

Let-7 family expression patterns in H, EA, and PA follicles. Detailed views of let-7a, let-7b, let-7c, let-7i and let-7g expression profiling in follicles undergoing atresia are presented in (A)–(E), respectively. Total RNA was extracted from the whole H, EA, and PA follicles for real-time PCR analysis. U6 snRNA was used as the normalization control. Data are presented as the mean ± SE. A one-way ANOVA test was used for multiple comparisons. P < 0.05 is shown as *, P < 0.01 is shown as **, and P < 0.001 is shown as ***.
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f4-molce-38-4-304: Let-7 family expression patterns in H, EA, and PA follicles. Detailed views of let-7a, let-7b, let-7c, let-7i and let-7g expression profiling in follicles undergoing atresia are presented in (A)–(E), respectively. Total RNA was extracted from the whole H, EA, and PA follicles for real-time PCR analysis. U6 snRNA was used as the normalization control. Data are presented as the mean ± SE. A one-way ANOVA test was used for multiple comparisons. P < 0.05 is shown as *, P < 0.01 is shown as **, and P < 0.001 is shown as ***.

Mentions: We discriminated H, EA and PA porcine ovary follicles by morphology and the ratio of Progesterone (P4)/17b-estradiol (E2) and extracted total RNA from each group. We then tested whether SYBR Green I real-time-PCR could quantify the let-7 family levels in these follicles. U6 snRNA was used as normalization control. The results are shown in Fig. 4. MiRNA let-7a, let-7b, let-7c and let-7i showed decreased abundance in EA and PA (Figs. 4A, 4B, 4C, 4D); however, let-7g was significantly increased in EA and PA relative to its expression in H (Fig. 4E). These results suggested that the expression patterns of the let-7 family members mostly correlated with the miRNA microarray data. The exception was let-7c, whose mature form did not show significantly reduced expression in EA relative to H based on our microarray data (Fig. 2C); however, stem-loop real-time-PCR showed that the relative abundance of mature let-7c was much lower in EA than in H (Fig. 4C).


Expression and preliminary functional profiling of the let-7 family during porcine ovary follicle atresia.

Cao R, Wu WJ, Zhou XL, Xiao P, Wang Y, Liu HL - Mol. Cells (2015)

Let-7 family expression patterns in H, EA, and PA follicles. Detailed views of let-7a, let-7b, let-7c, let-7i and let-7g expression profiling in follicles undergoing atresia are presented in (A)–(E), respectively. Total RNA was extracted from the whole H, EA, and PA follicles for real-time PCR analysis. U6 snRNA was used as the normalization control. Data are presented as the mean ± SE. A one-way ANOVA test was used for multiple comparisons. P < 0.05 is shown as *, P < 0.01 is shown as **, and P < 0.001 is shown as ***.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400304&req=5

f4-molce-38-4-304: Let-7 family expression patterns in H, EA, and PA follicles. Detailed views of let-7a, let-7b, let-7c, let-7i and let-7g expression profiling in follicles undergoing atresia are presented in (A)–(E), respectively. Total RNA was extracted from the whole H, EA, and PA follicles for real-time PCR analysis. U6 snRNA was used as the normalization control. Data are presented as the mean ± SE. A one-way ANOVA test was used for multiple comparisons. P < 0.05 is shown as *, P < 0.01 is shown as **, and P < 0.001 is shown as ***.
Mentions: We discriminated H, EA and PA porcine ovary follicles by morphology and the ratio of Progesterone (P4)/17b-estradiol (E2) and extracted total RNA from each group. We then tested whether SYBR Green I real-time-PCR could quantify the let-7 family levels in these follicles. U6 snRNA was used as normalization control. The results are shown in Fig. 4. MiRNA let-7a, let-7b, let-7c and let-7i showed decreased abundance in EA and PA (Figs. 4A, 4B, 4C, 4D); however, let-7g was significantly increased in EA and PA relative to its expression in H (Fig. 4E). These results suggested that the expression patterns of the let-7 family members mostly correlated with the miRNA microarray data. The exception was let-7c, whose mature form did not show significantly reduced expression in EA relative to H based on our microarray data (Fig. 2C); however, stem-loop real-time-PCR showed that the relative abundance of mature let-7c was much lower in EA than in H (Fig. 4C).

Bottom Line: In the current study, real-time quantitative polymerase chain reaction showed that the expression levels of let-7 family members in follicles and granulosa cells were similar to our microarray data, in which miRNAs let-7a, let-7b, let-7c, and let-7i were significantly decreased in early atretic and progressively atretic porcine ovary follicles compared with healthy follicles, while let-7g was highly expressed during follicle atresia.In addition, let-7 target genes were predicted and annotated by TargetScan, PicTar, gene ontology and Kyoto encyclopedia of genes and genomes pathways.Our data provide new insight into the association between the let-7 miRNA family in granulosa cell programmed death.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Most follicles in the mammalian ovary undergo atresia. Granulosa cell apoptosis is a hallmark of follicle atresia. Our previous study using a microRNA (miRNA) microarray showed that the let-7 microRNA family was differentially expressed during follicular atresia. However, whether the let-7 miRNA family members are related to porcine (Sus scrofa) ovary follicular apoptosis is unclear. In the current study, real-time quantitative polymerase chain reaction showed that the expression levels of let-7 family members in follicles and granulosa cells were similar to our microarray data, in which miRNAs let-7a, let-7b, let-7c, and let-7i were significantly decreased in early atretic and progressively atretic porcine ovary follicles compared with healthy follicles, while let-7g was highly expressed during follicle atresia. Furthermore, flow cytometric analysis and Hoechst33342 staining demonstrated that let-7g increased the apoptotic rate of cultured granulosa cells. In addition, let-7 target genes were predicted and annotated by TargetScan, PicTar, gene ontology and Kyoto encyclopedia of genes and genomes pathways. Our data provide new insight into the association between the let-7 miRNA family in granulosa cell programmed death.

No MeSH data available.


Related in: MedlinePlus