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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

Expression profiling of let-7 family in granulosa cells derived from in H, EA, and PA follicles. The relative abundance of let-7a, let-7b, let-7c, let-7i, let-7g in granulosa cells from H, EA and PA follicles are shown in (A) to (E), respectively. The granulosa cells, but not the whole follicles, were used for expression patterns analysis of let 7 family members using real-time PCR. U6 snRNA was used as normalization control. Data are presented as 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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f5-molce-38-4-304: Expression profiling of let-7 family in granulosa cells derived from in H, EA, and PA follicles. The relative abundance of let-7a, let-7b, let-7c, let-7i, let-7g in granulosa cells from H, EA and PA follicles are shown in (A) to (E), respectively. The granulosa cells, but not the whole follicles, were used for expression patterns analysis of let 7 family members using real-time PCR. U6 snRNA was used as normalization control. Data are presented as 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 extracted total RNA of granulosa cells from H, EA and PA follicles to test whether let-7 family were expressed in granulosa cells. The results were consistent with the expression pattern of let-7 family in follicles and microarray (Fig. 5). Previous studies showed that apoptosis triggered granulosa cell death and apoptotic granulosa cells increased in number during follicular atresia (Hughes and Gorospe, 1991; Sugimoto et al., 1998). Our results suggested that let-7 family were associated with the process of follicular apoptosis.


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)

Expression profiling of let-7 family in granulosa cells derived from in H, EA, and PA follicles. The relative abundance of let-7a, let-7b, let-7c, let-7i, let-7g in granulosa cells from H, EA and PA follicles are shown in (A) to (E), respectively. The granulosa cells, but not the whole follicles, were used for expression patterns analysis of let 7 family members using real-time PCR. U6 snRNA was used as normalization control. Data are presented as 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

f5-molce-38-4-304: Expression profiling of let-7 family in granulosa cells derived from in H, EA, and PA follicles. The relative abundance of let-7a, let-7b, let-7c, let-7i, let-7g in granulosa cells from H, EA and PA follicles are shown in (A) to (E), respectively. The granulosa cells, but not the whole follicles, were used for expression patterns analysis of let 7 family members using real-time PCR. U6 snRNA was used as normalization control. Data are presented as 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 extracted total RNA of granulosa cells from H, EA and PA follicles to test whether let-7 family were expressed in granulosa cells. The results were consistent with the expression pattern of let-7 family in follicles and microarray (Fig. 5). Previous studies showed that apoptosis triggered granulosa cell death and apoptotic granulosa cells increased in number during follicular atresia (Hughes and Gorospe, 1991; Sugimoto et al., 1998). Our results suggested that let-7 family were associated with the process of follicular apoptosis.

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