Cloning and Characterization of a Novel Oocyte-Specific Gene Encoding an F-Box Protein in Rainbow Trout (Oncorhynchus mykiss).

Wang, Lei; Tripurani, Swamy K.; Wanna, Warapond; Weber, Gregory M.; Rexroad, Caird E.; Yao, Jianbo

doi:10.1093/biolreprod/81.s1.349

Cited by 5 publications

(10 citation statements)

References 26 publications

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“…Electrophoresis and transfer were performed according to a previous study with minor modifications [45]. HEK293 cell were harvested with Pierce IP Lysis Buffer (Thermo Fisher Scientific, Waltham, MA), and 10 μl of cell lysate were mixed with an equal volume of Laemmli sample buffer (Bio-Rad Laboratories, Hercules, CA).…”

Section: Western Blot Analysismentioning

confidence: 99%

DNA methylation and miRNA-1296 act in concert to mediate spatiotemporal expression of KPNA7 during bovine oocyte and early embryonic development

Wang

Hand

et al. 2019

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Background Epigenetic regulation of oocyte-specific maternal factors is essential for oocyte and early embryonic development. KPNA7 is an oocyte-specific maternal factor, which controls transportation of nuclear proteins important for early embryonic development. To elucidate the epigenetic mechanisms involved in the controlled expression of KPNA7, both DNA methylation associated transcriptional silencing and miRNA-mediated mRNA degradation of KPNA7 were examined. Results Comparison of DNA methylation profiles at the proximal promoter of KPNA7 gene between oocyte and 6 different somatic tissues identified 3 oocyte-specific differentially methylated CpG sites. Expression of KPNA7 mRNA was reintroduced in bovine kidney-derived CCL2 cells after treatment with the methylation inhibitor, 5-aza-2-deoxycytidine (5-Aza). Analysis of the promoter region of KPNA7 gene in CCL2 cells treated with 5-Aza showed a lighter methylation rate in all the CpG sites. Bioinformatic analysis predicted 4 miRNA-1296 binding sites in the coding region of KPNA7 mRNA. Ectopic co-expression of miRNA-1296 and KPNA7 in HEK293 cells led to reduced expression of KPNA7 protein. Quantitative real time PCR analysis revealed that miRNA-1296 is expressed in oocytes and early stage embryos, and the expression reaches a peak level in 8-cell stage embryos, coincident with the time of embryonic genome activation and the start of declining of KPNA7 expression. Conclusions These results suggest that DNA methylation may account for oocyte-specific expression of KPNA7, and miRNA-1296 targeting the coding region of KPNA7 is a potential mechanism for KPNA7 transcript degradation during the maternal-to-zygotic transition.

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Section: Western Blot Analysismentioning

confidence: 99%

DNA methylation and miRNA-1296 act in concert to mediate spatiotemporal expression of KPNA7 during bovine oocyte and early embryonic development

Wang

Hand

et al. 2019

Preprint

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“…Electrophoresis and transfer were performed according to a previous study with minor modifications [45].…”

Section: Western Blot Analysismentioning

confidence: 99%

DNA methylation and miRNA-1296 act in concert to mediate spatiotemporal expression of KPNA7 during bovine oocyte and early embryonic development

Wang

Hand

et al. 2019

Preprint

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Epigenetic regulation of oocyte-specific maternal factors is essential for oocyte and early embryonic development. KPNA7 is an oocyte-specific maternal factor, which controls transportation of nuclear proteins important for early embryonic development. To elucidate the epigenetic mechanisms involved in the controlled expression of KPNA7, both DNA methylation associated transcriptional silencing and miRNA-mediated mRNA degradation of KPNA7 were examined. Results Comparison of DNA methylation profiles at the proximal promoter of KPNA7 gene between oocyte and 6 different somatic tissues identified 3 oocyte-specific differentially methylated CpG sites. Expression of KPNA7 mRNA was reintroduced in bovine kidney-derived CCL2 cells after treatment with the methylation inhibitor, 5-aza-2-deoxycytidine (5-Aza). Analysis of the promoter region of KPNA7 gene in CCL2 cells treated with 5-Aza showed a lighter methylation rate in all the CpG sites. Bioinformatic analysis predicted 4 miRNA-1296 binding sites in the coding region of KPNA7 mRNA. Ectopic co-expression of miRNA-1296 and KPNA7 in HEK293 cells led to reduced expression of KPNA7 protein. Quantitative real time PCR analysis revealed that miRNA-1296 is expressed in oocytes and early stage embryos, and the expression reaches a peak level in 8-cell stage embryos, coincident with the time of embryonic genome activation and the start of declining of KPNA7 expression. Conclusions These results suggest that DNA methylation may account for oocyte-specific expression of KPNA7, and miRNA-1296 targeting the coding region of KPNA7 is a potential mechanism for KPNA7 transcript degradation during the maternal-to-zygotic transition.

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“…These data indicate that Figla plays a critical role in activation of oocyte development and repression of testis development to maintain the sex-specific germ cell development. In cattle, Figla shows similar expression profile as in other mammals, which is restricted to gonadal tissues and appears as early as primordial follicle stage (Tripurani, et al 2013) were decreased after embryonic genome activation suggesting miRNA212 might be a maternal origin factor involved in maternal transcript degradation during the MZT.…”

Section: Figla: Factor In Germline Alphamentioning

confidence: 58%

“…and Figla 3'UTR, and studies on in vitro cultured (IVC) bovine embryos revealed that miRNA212 mimic inhibits Figla protein expression (Tripurani, et al 2013). These studies indicate that Figla is a critical factor for follicular and embryo development and miRNA212 is a potential post-transcriptional regulator of Figla during the MZT in cattle.…”

Section: Figla: Factor In Germline Alphamentioning

confidence: 83%

“…However, studies of egg-specific genes in fish are limited. Based on analysis of EST sequences from a rainbow trout egg cDNA library, we have previously identified and characterized three novel egg-specific genes in rainbow trout that include oorp-t (Ramachandra et al 2007), rtgst-1 (Qiu et al 2008) and fbxoo (Wang et al 2013). In this study, we report the characterization of another egg-specific gene, kpna7, in rainbow trout.…”

Section: Discussionmentioning

confidence: 95%

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Functional Analysis and Transcriptional Regulation of Oocyte-Specific Nuclear Transporter KPNA7

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Functional Analysis and Transcriptional Regulation of Oocyte-Specific nuclear transporter KPNA7 Lei Wang The finely orchestrated development and maturation of the oocytes has been the focus of many studies in which oocyte-specific genes essential for folliculogenesis and early embryonic development have been identified. Studies using knockout mice models of Nobox, Sohlh1, Sohlh2, Figla and Lhx8 have revealed that these oocyte specific transcriptional factors form networks in which they regulate the expression of each other and downstream targets. However, little is known about the mechanisms that control the nuclear transport of these essential factors in oocytes and early embryos. The nuclear envelope separates the nuclear and cytoplasmic compartment in the cell and uncouples transcription and translation; therefore, nucleo-cytoplasmic transport is necessary and highly regulated in eukaryotic cells. The soluble proteins that play key roles in mediating the bidirectional trafficking of macromolecules across the nuclear envelope are collectively called Karyopherins (KPNs), with those involved in nuclear import referred to as KPNAs. The first oocyte-specific KPNA gene, KPNA7, was discovered in bovine oocytes; later the homologous genes were found in mice, human and pig. Bovine KPNA7 is highly expressed in germinal vesicle (GV) and metaphase II oocytes, as well as in early stage embryos collected before embryonic genome activation, but is barely detectable in morula and blastocyst stage embryos after the maternal to embryonic transmission. Female mice lacking KPNA7 gene are affiliated with reproductive reduction and sex imbalance by inducing preferential fetal lethality. Induced abnormalities of gene expression (Dppa2, Dppa4 and Piwil2) and epigenetic modifications (down-regulation of histone H3K27me3) are also part of the Kpna7 knock out effects. Although it is known that KPNA7 is a maternal transcript critical for development in mammals, little is known about mechanism of regulation its expression and the nuclear translocation function of KPNA7 protein in bovine embryos or the expression and functions of KPNA7 in rainbow trout.

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Cloning and Characterization of a Novel Oocyte-Specific Gene Encoding an F-Box Protein in Rainbow Trout (Oncorhynchus mykiss).

Cited by 5 publications

References 26 publications

DNA methylation and miRNA-1296 act in concert to mediate spatiotemporal expression of KPNA7 during bovine oocyte and early embryonic development

DNA methylation and miRNA-1296 act in concert to mediate spatiotemporal expression of KPNA7 during bovine oocyte and early embryonic development

DNA methylation and miRNA-1296 act in concert to mediate spatiotemporal expression of KPNA7 during bovine oocyte and early embryonic development

Functional Analysis and Transcriptional Regulation of Oocyte-Specific Nuclear Transporter KPNA7

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