Expression and Subcellular Localization of GSE Protein in Germ Cells and Preimplantation Embryos

Mizuno, Satoshi; Sono, Youhei; Matsuoka, Toshifumi; Matsumoto, Kazuya; Saeki, Kazuhiro; Hosoi, Yoshihiko; Fukuda, Aisaku; Morimoto, Yoshiharu; Iritani, Akira

doi:10.1262/jrd.18005

Cited by 17 publications

(27 citation statements)

References 23 publications

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“…The procedures for in vitro fertilization were essentially those reported previously [22]. In brief, spermatozoa were collected from the cauda epididymis of a male mouse.…”

Section: In Vitro Fertilization and Culturementioning

confidence: 99%

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Inhibition of the Ubiquitin-proteasome System Leads to Delay of the Onset of ZGA Gene Expression

et al. 2010

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Abstract. In mammalian oocytes, the ubiquitin-proteasome system (UPS) is suggested to play important roles in oocyte meiosis resumption, spindle assembly, polar body emission and pronuclear formation by regulating cyclin B1 degradation. However, little is known about the direct relationship between zygotic gene activation (ZGA) and degradation of maternal proteins. Here, we investigated the role of the UPS in the onset of ZGA in early mouse embryos. First, we found degradation of cyclin B1 protein in fertilized oocytes at 1 hpi by western blot analysis and used these oocytes throughout this study. Subsequently, we determined optimal experimental conditions for transient inhibition of proteasomal activity by specific and reversible proteasomal inhibitor MG132 in the G1 phase of the first cell cycle. Under the selected optimal conditions, we subjected transient MG132-treated embryos to reverse transcription (RT)-PCR analysis of expression of four ZGA genes, i.e., the hsp70.1, MuERV-L, eif-1a and zscan4d genes. As a result, we found that onset of expression of the four examined ZGA genes was delayed in both normally developed 2-cell embryos and arrested 1-cell embryos. Our results indicate that proteasomal degradation of proteins by the UPS plays a pivotal role in the molecular mechanisms of ZGA in early mouse embryos. Key words: Early embryo, MG132, Mouse, Ubiquitin-proteasome system, Zygotic gene activation (J. Reprod. Dev. 56: [655][656][657][658][659][660][661][662][663] 2010) he proteasome is a large protein complex for degradation of most intracellular proteins in eukaryotic cells [1,2]. For proteasomal degradation, the substrate proteins are required for tagging by ubiquitin. Generally, the ubiquitin-proteasome system (UPS) is involved in a wide variety of biological processes, including DNA repair, apoptosis, immune response, signal transduction, transcription, metabolism, protein quality control and developmental programs [3,4].In mammalian oocytes, the UPS is suggested to play important roles in oocyte meiosis resumption, spindle assembly, polar body emission and pronuclear formation by regulating cyclin B1 degradation [5][6][7]. Ubiquitin enzyme E1 [8], E2 [9] and E3 [10] are active during mammalian spermatogenesis, suggesting that the UPS may play roles during spermatogenesis [11]. In fertilization, ubiquitination of the sperm mitochondrial membranes may assist in destruction of paternal mitochondria at fertilization for promoting the maternal inheritance of mitochondrial DNA in mammals [11,12].In mouse embryos, the onset of zygotic gene activation (ZGA) has been shown to occur after fertilization and is a critical event that governs the maternal-to-zygotic transition (MZT) for embryonic development [13]. The correct regulation of the onset of ZGA is an important process for remodeling of an oocyte to a totipotent zygote. Recently, several studies have implicated the UPS in the regulation of transcription [14][15][16][17]. These observations led us to investigate a potential link between the UPS and ...

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“…The procedures for in vitro fertilization were essentially those reported previously [22]. In brief, spermatozoa were collected from the cauda epididymis of a male mouse.…”

Section: In Vitro Fertilization and Culturementioning

confidence: 99%

“…The procedures for western blot analysis were essentially the same as those reported previously [22,24,25]. In brief, protein extracts were prepared from MII oocytes and fertilized oocytes at 1 hpi, at which time the second polar body (2nd PB) was already extruded from metaphase II oocytes after fertilization.…”

Section: Western Blot Analysismentioning

confidence: 99%

Inhibition of the Ubiquitin-proteasome System Leads to Delay of the Onset of ZGA Gene Expression

et al. 2010

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“…GSE is a maternal protein which is expressed up from the blastocyst stage to the immature oocyte, although the abundance of this protein decreases from the blastocyst stage (90,91) . In the zygote GSE protein is found preferentially in the male pronucleus attached to chromatin (91) .…”

Section: Efecto Materno Sobre La Epigénesismentioning

confidence: 99%

“…GSE es una proteína materna que se expresa desde el ovocito inmaduro hasta la etapa de blastocisto, aunque la abundancia de esta proteína disminuye a partir de este estadio (90,91) . En el cigoto la proteina GSE se encuentra de forma preferencial en el pronúcleo masculino unida a la cromatina (91) .…”

Section: Efecto Materno Sobre La Epigénesisunclassified

Efectos de los RNAm maternos sobre la maduración del ovocito y el desarrollo embrionario temprano en mamíferos

Burrola-Barraza¹,

González-Rodríguez

2015

Rev. Mex. Cienc. Pecu.

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Efectos de los RNAm maternos sobre la maduración del ovocito y el desarrollo embrionario temprano en mamíferos. RevisiónEffects of the maternal mRNA on the maturation of the oocyte and early embryonic development in mammals. Review M. Eduviges Burrola-Barraza a , Everardo González-Rodríguez a RESUMENObtener ovocitos madurados in vitro, que sean competentes para fertilizarse y originar un porcentaje superior al 50 % de blastocitos viables, es una de las principales metas que existen en el desarrollo in vitro de embriones bovinos. Es por ello necesario entender los procesos celulares, que desembocan en la maduración del ovocito durante la foliculogénesis y su subsecuente transición al embrión. La transición del ovocito al embrión es un proceso complejo que involucra la inactivación genómica del ovocito y la activación del genoma embrionario. Este proceso se da en bovinos en la etapa de 8 a 16 células y presenta una degradación selectiva de RNAm maternos, que fueron almacenados durante la ovogénesis y son la fuente para la codificación de proteínas en las etapas iníciales del desarrollo embrionario. La acción de los RNAm maternos es clave para que la activación del genoma embrionario se realice en tiempo y forma adecuados. Entender la participación de estos transcritos en la activación del genoma embrionario, es esencial para esclarecer los procesos celulares que fallan en los protocolos de maduración de ovocitos in vitro. Es por esto que el objetivo de esta revisión fue recopilar la información de los principales RNAm maternos que han sido identificados tanto en el modelo murino como el bovino, esto con el fin de integrar el conocimiento relacionado con los procesos genómicos que permiten el desarrollo del embrión en el bovino, y diseñar nuevas estrategias que permitan mejorar los protocolos de fertilización in vitro.PALABRAS CLAVE: Ovocito, Activación genoma embrionario, Bovino. ABSTRACTGet in vitro matured oocytes that could be competent for in vitro fertilization with a result of 50 % viable blastocysts, is one of the main problems in the in vitro development of bovine embryos. For these reason, is necessary to understand the cellular processes that lead to the maturation of the oocyte during folliculogenesis and subsequent transition to the embryo. The transition from oocyte to embryo is a complex process that involves genomic inactivation of the oocyte and embryonic genome activation. This process occurs in cattle in the stage of 8 to 16 cells where there are a selective degradation of maternal mRNA, which were stored during oogenesis and are the source of protein in the early stages of embryonic development. The action of maternal mRNA is a key for that embryonic genome activation takes place in form and time. Understanding the involvement of these transcripts in embryonic genome activation is essential to elucidate the cellular processes falling in oocyte maturation in vitro protocols. The aim of this review was to gather information from maternal genes that have been isolated both in the murine model and c...

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“…The procedures were performed according to previously reported methods [19][20][21][22]. In brief, the proteins obtained as mentioned above were subjected to SDS-PAGE.…”

Section: Western Blot Analysismentioning

confidence: 99%

Proteomic Analysis of the Mouse Ovary in Response to Two Gonadotropins, Follicle-Stimulating Hormone and Luteinizing Hormone

Satoh

Tokoro

Ikegami

et al. 2009

Journal of Reproduction and Development

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Abstract. Functional and structural changes in the mammalian ovary are coordinately regulated by the pituitary glycoprotein hormones, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), leading to follicular development, ovulation and transformation of follicles into corpus lutea. To investigate protein profiles during these processes of the mouse ovarian cycle, we applied combined methods (two-dimensional gel electrophoresis for separation and visualization of proteins plus matrix laser desorption/ionization time-of-flight mass spectrometry [MALDI-TOF/MS] analysis for protein identification) for comparative proteomic analysis using immature mice at 3 weeks of age. Protein profiles were obtained from proteins extracted from intact ovaries that had been collected from pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-primed immature mice at 0 (no PMSG), 24 and 48 h post PMSG, as well as at 10 and 20 h post hCG. The results showed that 1028 common protein spots were found in representative gels that had been separated in the 3 to 11 pH range and the 15-200 kDa range, 253 protein spots (24.6%) of which were differentially expressed (p<0.05) during the mouse ovarian cycle. Of these 253 protein spots, 99 were identified by MALDI-TOF/MS. This comparative proteomic approach to identifying proteins that were potentially involved in the complex process of the ovarian cycle could contribute to our understanding of the molecular basis of functional and structural changes in the ovary in response to gonadotropins. Furthermore, the interesting ovarian proteins identified in this study may eventually serve as diagnostic biomarker candidates of ovarian function. Key words: Follicle stimulating hormone, Luteinizing hormone, Ovary, Proteome, Two-dimensional gel electrophoresis (J. Reprod. Dev. 55: [316][317][318][319][320][321][322][323][324][325][326] 2009) omplex biological processes in the mammalian ovary, such as follicular development, oocyte maturation, oocyte meiosis, ovulation and corpus luteum formation and demise, are coordinately regulated by autocrine, paracrine and endocrine factors of the hypothalamic-pituitary-ovarian axis [1][2][3]. Specifically, follicle-stimulating hormone (FSH) is a major promoter for orchestrating follicular development and differentiation in the granulosa cells of preovulatory follicles [4][5][6]. Luteinizing hormone (LH) plays a key role in initiation of the ovulatory process of preovulatory follicles by activating multiple cellular signaling pathways [5,7]. Detailed biological changes in gene expression and follicular structure occur due to the overlapping control of FSH and LH, and the theca, granulosa, cumulus and oocyte compartments of the ovarian follicle are interdependently regulated by them. Moreover, the universal and local actions of these gonadotropins are coordinated with signals from the oocyte to promote ovulation under multipartite control, facilitating synchronization of oocyte maturation with follicle rupture and resulting in select...

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Expression and Subcellular Localization of GSE Protein in Germ Cells and Preimplantation Embryos

Cited by 17 publications

References 23 publications

Inhibition of the Ubiquitin-proteasome System Leads to Delay of the Onset of ZGA Gene Expression

Inhibition of the Ubiquitin-proteasome System Leads to Delay of the Onset of ZGA Gene Expression

Efectos de los RNAm maternos sobre la maduración del ovocito y el desarrollo embrionario temprano en mamíferos

Proteomic Analysis of the Mouse Ovary in Response to Two Gonadotropins, Follicle-Stimulating Hormone and Luteinizing Hormone

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