Identification and expression analysis of microRNAs in medaka gonads

Qiu, Weiwei; Zhu, Yefei; Wu, Yun; Yuan, Can‐Can; Chen, Kerang; Li, Mingyou

doi:10.1016/j.gene.2017.12.062

Cited by 44 publications

(38 citation statements)

References 36 publications

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“…Consistently, 15 recent transcriptomic studies in zebrafish reported either an increased expression of 16 miR-202-5p during folliculogenesis or an increased expression in developing oocytes 17 and unfertilized eggs (48)(49). Yet, it has also been claimed that miR-202 could by 18 expressed only in early medaka oocytes and absent at later stages, based on ISH 19 experiments (16). Since our ISH did no reveal any expression of miR-202-5p above 20 background levels in early oocytes, we presume that it is more likely accumulated 21 during vitellogenesis in vitellogenic and/or post-vitellogenic oocytes, as suggested by 22 the detectable expression in unfertilized eggs, 23…”

Section: Discussionmentioning

confidence: 50%

“…Females lacking miR-202 produce eggs that cannot be fertilized 16 In this study, we paid special attention to the severe phenotype displayed by miR-202 17 KO females. The overall reproductive success of mutant females was reduced to 0.83 or, 18 even more, to O viable eggs per clutch, compared to 15.3 viable eggs per clutch for wild-19 type females.…”

Section: Discussionmentioning

confidence: 99%

“…Mir-202, a major player rather than a fine modulator 16 As discussed above, a drastic reduction of the reproductive success was observed for 17 both male and female medaka lacking miR-202, including a reduced female fecundity 18 and the production of poor quality eggs that cannot be fertilized. This phenotype was 19 confirmed using two mutant lines bearing different INDELs obtained with the same 20 guide RNA.…”

Section: Mir-202 Knock-out Impairs Female Meiosismentioning

confidence: 96%

“…More specifically, expression and regulation of miR-17 and miR-430b in the ovary 17 suggest a role in follicular development and oocyte maturation. Large scale differential 18 transcriptomic analyses were also performed during sex differentiation, gonadal 19 development or vitellogenesis in various fish species, including rainbow trout (9), 20 zebrafish (10)(11), Atlantic halibut (12), yellow cat fish (13), Tilapia (14) (15) and 21 medaka (16) (17). The identification of differentially expressed miRNAs through the 22 different stages of the reproductive cycle is consistent with a participation of miRNAs in 23 oogenesis and spermatogenesis.…”

Section: Introductionmentioning

confidence: 99%

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MicroRNA-202 (miR-202) controls female fecundity by regulating medaka oogenesis

Bugeon

Bouchareb

Henry

et al. 2018

Preprint

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Female gamete production relies on coordinated molecular and cellular processes that occur in the ovary throughout oogenesis. In fish, as in other vertebrates, these processes have been extensively studied both in terms of endocrine/paracrine regulation and protein expression and activity. The role of small non-coding RNAs in the regulation of animal reproduction remains however largely unknown and poorly investigated, despite a growing interest for the importance of miRNAs in a wide variety of biological processes. Here, we analyzed the role of miR-202, a miRNA predominantly expressed in male and female gonads in several vertebrate species. We studied its expression in the medaka ovary and generated a mutant line (using CRISPR/Cas9 genome engineering) to determine its importance for reproductive success with special interest for egg production. Our results show that miR-202-5p is the biologically active form of the miRNA and that it is expressed in granulosa cells and in the unfertilized egg. The knock out (KO) of miR-202 resulted in a strong phenotype both in terms of number and quality of eggs produced. Mutant females exhibited either no egg production or produced a drastically reduced number of eggs that could not be fertilized, ultimately leading to no reproductive success. We quantified the size distribution of the oocytes in the ovary of KO females and performed a genome-wide transcriptomic analysis approach to identified dysregulated molecular pathways. Together, cellular and molecular analyses indicate that lack of miR-202 impairs the early steps of oogenesis/folliculogenesis and decreases the number of large (i.e. vitellogenic) follicles, ultimately leading to dramatically reduced female fecundity. This study sheds new light on the regulatory mechanisms that control the early steps of follicular development and provides the first in vivo functional evidence that an ovarian-predominant microRNA may have a major role in female reproduction.Author summaryThe role of small non-coding RNAs in the regulation of animal reproduction remains poorly investigated, despite a growing interest for the importance of miRNAs in a wide variety of biological processes. Here, we analyzed the role of miR-202, a miRNA predominantly expressed in gonads in vertebrate. We studied its expression in the medaka ovary and knocked out the miR-202 genes to study its importance for reproductive success. We showed that the lack of miR-202 results in the sterility of both females and males. In particular, it lead to a drastic reduction of both the number and the quality of eggs produced by females. Mutant females exhibited either no egg production or produced a drastically reduced number of eggs that could not be fertilized, ultimately leading to no reproductive success. Quantitative histological and molecular analyses indicated that miR-202 KO impairs oocyte development and is also associated with the dysregulation of many genes that are critical for reproduction. This study sheds new light on the regulatory mechanisms that control oogenesis and provides the first in vivo functional evidence that an ovarian-predominant microRNA may have a major role in female reproduction.

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Section: Discussionmentioning

confidence: 50%

Section: Discussionmentioning

confidence: 99%

Section: Mir-202 Knock-out Impairs Female Meiosismentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

MicroRNA-202 (miR-202) controls female fecundity by regulating medaka oogenesis

Bugeon

Bouchareb

Henry

et al. 2018

Preprint

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“…One of our previous studies of testis transcriptional pro le revealed numerous differentially expressed genes (DEGs) and important biological pathways were signi cantly correlated to mammalian reproduction between Meishan and Duroc boars at 20, 75, and 270 days of age [17]. It is interesting to further investigate how miRNAs are involved in regulating sexual function development by ne-turning gene expression [18]. Chinese Meishan pigs are one of the most proli c pig breeds in the world and reach puberty at a relatively younger age (56-84 days) than conventional boars (120-180 days) [19].…”

Section: Introductionmentioning

confidence: 99%

Integrated analysis of miRNA and mRNA expression profiles in testes of Duroc and Meishan boars

Ding

Liu

Zhou

et al. 2020

Preprint

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Background: MicroRNAs (miRNAs) are small non-coding RNAs playing vital roles in regulating posttranscriptional gene expression. Elucidating the expression regulation of miRNAs underlying pig testis development will contribute to a better understanding of boar fertility and spermatogenesis. Results: In this study, miRNA expression profile was investigated in testes of Duroc and Meishan boars at 20, 75, and 270 days of age by high-throughput sequencing. Forty-five differentially expressed miRNAs were identified from testes of Duroc and Meishan boars before and after puberty. Integrated analysis of miRNA and mRNA profiles predicted many miRNA-mRNA pairs. Gene ontology and biological pathway analyses revealed that predicted target genes of ssc-mir-423-5p, ssc-mir-34c, ssc-mir-107, ssc-165 mir-196b-5p, ssc-mir-92a, ssc-mir-320, ssc-mir-10a-5p, and ssc-mir-181b were involved in sexual reproduction, male gamete generation, and spermatogenesis, and GnRH, Wnt, and MAPK signaling pathway. Four significantly differentially expressed miRNAs and their predicted target genes were validated by quantitative real-time polymerase chain reaction, and phospholipase C beta 1 ( PLCβ1) gene was verified to be a target of ssc-mir-423-5p . Conclusions: This study provides an insight into the functional roles of miRNAs in testis development and spermatogenesis and offers useful resources for understanding differences in sexual function development caused by the change in miRNAs expression between Duroc and Meishan boars.

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