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A novel functional role for the oocyte-specific transcription factor newborn ovary homeobox (NOBOX) during early embryonic development in cattle.

ABSTRACT: Newborn ovary homeobox (NOBOX) is an oocyte-specific transcription factor essential for folliculogenesis and expression of many germ cell-specific genes in mice. Here we report the characterization of the bovine NOBOX gene and its role in early embryogenesis. The cloned cDNA for bovine NOBOX contains an open reading frame encoding a protein of 500 amino acids with a conserved homeodomain. mRNA for NOBOX is preferentially expressed in ovaries and undetectable by RT-PCR in somatic tissues examined. NOBOX protein is present in oocytes throughout folliculogenesis. NOBOX is expressed in a stage-specific manner during oocyte maturation and early embryonic development and of maternal origin. Knockdown of NOBOX in early embryos using small interfering RNA demonstrated that NOBOX is required for embryonic development to the blastocyst stage. Depletion of NOBOX in early embryos caused significant down-regulation of genes associated with transcriptional regulation, signal transduction, and cell cycle regulation during embryonic genome activation. In addition, NOBOX depletion in early embryos reduced expression of pluripotency genes (POU5F1/OCT4 and NANOG) and number of inner cell mass cells in embryos that reached the blastocyst stage. This study demonstrates that NOBOX is an essential maternal-derived transcription factor during bovine early embryogenesis, which functions in regulation of embryonic genome activation, pluripotency gene expression, and blastocyst cell allocation.

SUBMITTER: Tripurani SK

PROVIDER: S-EPMC3040056 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature

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A novel functional role for the oocyte-specific transcription factor newborn ovary homeobox (NOBOX) during early embryonic development in cattle.

Tripurani Swamy K SK Lee Kyung-Bon KB Wang Lei L Wee Gabbine G Smith George W GW Lee Young S YS Latham Keith E KE Yao Jianbo J

Endocrinology 20101230 3

Newborn ovary homeobox (NOBOX) is an oocyte-specific transcription factor essential for folliculogenesis and expression of many germ cell-specific genes in mice. Here we report the characterization of the bovine NOBOX gene and its role in early embryogenesis. The cloned cDNA for bovine NOBOX contains an open reading frame encoding a protein of 500 amino acids with a conserved homeodomain. mRNA for NOBOX is preferentially expressed in ovaries and undetectable by RT-PCR in somatic tissues examined ...[more]

PMID: 21193554

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Project description:BackgroundOocyte-derived maternal RNAs drive early embryogenesis when the newly formed embryo is transcriptionally inactive. Recent studies in zebrafish have identified the role of microRNAs during the maternal-to-embryonic transition (MET). MicroRNAs are short RNAs that bind to the 3' UTR of target mRNAs to repress their translation and accelerate their decay. Newborn ovary homeobox gene (NOBOX) is a transcription factor that is preferentially expressed in oocytes and essential for folliculogenesis in mice. NOBOX knockout mice are infertile and lack of NOBOX disrupts expression of many germ-cell specific genes and microRNAs. We recently reported the cloning and expression of bovine NOBOX during early embryonic development and our gene knockdown studies indicate that NOBOX is a maternal effect gene essential for early embryonic development. As NOBOX is a maternal transcript critical for development and NOBOX is depleted during early embryogenesis, we hypothesized that NOBOX is targeted by microRNAs for silencing and/or degradation.ResultsUsing an algorithm "MicroInspector", a potential microRNA recognition element (MRE) for miR-196a was identified in the 3' UTR of the bovine NOBOX mRNA. Expression analysis of miR-196a in bovine oocytes and during early embryonic development indicated that it is expressed both in oocytes and embryos and tends to increase at the four-cell and eight-cell stages. Ectopic expression of NOBOX and miR-196a in HeLa cells inhibited the expression of NOBOX protein compared to the control cells without miR-196a. Similarly, the activity of a luciferase construct containing the entire 3' UTR of bovine NOBOX was suppressed, and the regulation was abolished by mutations in the miR-196a binding site indicating that the predicted MRE is critical for the direct and specific binding of miR-196a to the NOBOX mRNA. Furthermore, ectopic expression of miR-196a mimic in bovine early embryos significantly reduced the NOBOX expression at the both mRNA and protein levels.ConclusionCollectively, our results demonstrate that miR-196a is a bona fide negative regulator of NOBOX during bovine early embryogenesis.

Dataset Information

A novel functional role for the oocyte-specific transcription factor newborn ovary homeobox (NOBOX) during early embryonic development in cattle.

Publications

A novel functional role for the oocyte-specific transcription factor newborn ovary homeobox (NOBOX) during early embryonic development in cattle.

Similar Datasets

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