Meiotic Resumption of Porcine Immature Oocytes is Prevented by Ooplasmic Gs.ALPHA. Functions

Morikawa, Marie; Seki, Mami; Kume, Sachi; Endo, Toshihiko; Nishimura, Yukio; Kanô, Kiyoshi; Naito, Kunihiko

doi:10.1262/jrd.19055

Cited by 11 publications

(12 citation statements)

References 38 publications

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“…In the present study, we confirmed, in a chemically defined gonadotropin-free medium, that dibutyryl cAMP could regulate the meiotic resumption of porcine oocytes more effectively than EGFfamily peptides alone when COCs are exposed to dibutyryl cAMP or EGF-family peptides during the first 20 h of IVM. Intracellular cAMP appears to regulate meiotic resumption of oocytes through the signaling pathways involved in maturation promoting factor (MPF) activation [38].In the current study, we also found a synergistic effect of dibutyryl cAMP and EGF-family peptides not only on meiotic resumption, but also on the regulation of meiotic progression to the metaphase-II stage in porcine oocytes. In particular, exposure of COCs to a combination of dibutyryl cAMP with EGF plus amphiregulin during the first 20 h of IVM improved both the incidences of meiotic resumption and progression to the metaphase-II stage to the same levels as oocytes treated with gonadotropins and dibutyryl cAMP.…”

supporting

confidence: 68%

Section: Discussionmentioning

confidence: 99%

“…Intracellular cAMP appears to regulate meiotic resumption of oocytes through the signaling pathways involved in maturation promoting factor (MPF) activation [38].…”

Section: Discussionmentioning

confidence: 99%

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Successful Piglet Production in a Chemically Defined System for In-vitro Production of Porcine Embryos: Dibutyryl Cyclic AMP and Epidermal Growth Factor-family Peptides Support In-vitro Maturation of Oocytes in the Absence of Gonadotropins

Akaki

Yoshioka

Noguchi

et al. 2009

Journal of Reproduction and Development

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Abstract.To induce meiotic resumption of porcine oocytes, it is thought to be necessary to expose the cumulus-oocyte complexes (COCs) to gonadotropins during in-vitro maturation (IVM). However, the detailed mechanism of meiotic resumption by gonadotropins is still unknown, and successful piglet production has not been reported by using oocytes matured in gonadotropin-free media and fertilized in vitro. The present study was undertaken to examine the combinational effects of epidermal growth factor (EGF)-family members and dibutyryl cyclic AMP (cAMP) in a chemically defined medium on IVM of porcine oocytes and the developmental competence following in vitro fertilization (IVF). The basic IVM medium was a chemically defined medium, modified porcine oocyte medium (mPOM). Supplementation of the IVM medium with 10 or 1000 ng/ml EGF, amphiregulin and betacellulin during the whole IVM period, except for 10 ng/ml amphiregulin, increased the percentage of oocytes maturing to the metaphase-II stage. When COCs were exposed to both dibutyryl cAMP and EGF-family members during the first 20-h of IVM and then culture was continued in the absence of EGF-family members and dibutyryl cAMP, the incidence of metaphase-II oocytes was significantly increased and was not different from that of oocytes cultured in a standard IVM system with gonadotropins. The developmental competence of the oocytes to the blastocyst stage following IVF was no different from that of control oocytes matured with gonadotropins. When these blastocysts were transferred into the uterine horn of three recipients, all of gilts became pregnant and delivered a total of 11 piglets. These observations indicate that supplementation of a chemically defined maturation medium with EGF-family members and dibutyryl cAMP during the first 20 h of IVM can support well the meiotic progress and developmental competence of porcine oocytes. Key words: Cyclic AMP (cAMP), Epidermal growth factor (EGF), In vitro maturation, Oocyte, Pig (J. Reprod. Dev. 55: [446][447][448][449][450][451][452][453] 2009) uccessful in-vitro production of porcine embryos has the potential to provide an effective system for mass production of embryos for better understanding of the physiology of embryonic development and animal reproductive technologies, including production of transgenic and/or cloned animals [1,2]. Relatively efficient systems for in-vitro production of porcine embryos have already been developed, but they contain materials derived from organisms, such as porcine follicular fluid and bovine serum albumin, in the culture media [3,4]. Chemically defined media, which only known chemicals, but none derived from animals, yeast or plants, should be useful in clarifying the mechanism of interactive communication between the oocyte and the surrounding cumulus cells during meiosis, the oocyte reaction following sperm penetration and early development. Furthermore, application of a defined system for in-vitro embryo production would be good for a quality control against the risk of viral c...

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supporting

confidence: 68%

Section: Discussionmentioning

confidence: 99%

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Successful Piglet Production in a Chemically Defined System for In-vitro Production of Porcine Embryos: Dibutyryl Cyclic AMP and Epidermal Growth Factor-family Peptides Support In-vitro Maturation of Oocytes in the Absence of Gonadotropins

Akaki

Yoshioka

Noguchi

et al. 2009

Journal of Reproduction and Development

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“…The activities of MPF were evaluated on the basis of the histone H1 kinase activity as described in previous reports [15].…”

Section: Mpf Activity Assaymentioning

confidence: 99%

Analyses of the Regulatory Mechanism of Porcine WEE1B: The Phosphorylation Sites of Porcine WEE1B and Mouse WEE1B Are Different

Shimaoka

Nishimura

Kanô

et al. 2011

Journal of Reproduction and Development

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Abstract. WEE1B, an oocyte-specific kinase, phosphorylates the CDC2 inhibitory site and maintains the meiotic arrest of oocytes at the first meiotic prophase in several mammalian species. However, the molecular mechanisms controlling WEE1B activity have not been fully examined in species other than mice. In the present study, we analyzed the regulation mechanisms of porcine WEE1B (pWEE1B), focusing on the cAMP-dependent protein kinase (PKA) phosphorylation site and intracellular localization. As the PKA phosphorylation site in mouse WEE1B (mWEE1B) was not conserved in pWEE1B, we predicted that four serine residues would be phosphorylatable by PKA in pWEE1B (Ser77, Ser118, Ser133 and Ser149) and constructed FLAG-tagged replaced-pWEE1Bs, in which each of the PKAphosphorylatable serines was mutated into a non-phosphorylatable alanine. We injected one of their mRNAs into porcine immature oocytes and found that the Ser77-replaced pWEE1B lost the WEE1B function, whereas the wild-type and other replaced-pWEE1Bs could maintain the meiotic arrest of oocytes. Next, the localization of pWEE1B was examined by immunohistochemistry, and exclusive nuclear localization was revealed in the fully grown oocytes. We generated a nuclear localization signal (NLS)-deleted pWEE1B (ΔNLS-pWEE1B) and then overexpressed it in porcine immature oocytes. We found that ΔNLS-pWEE1B was distributed uniformly in the cytoplasm and could not maintain the meiotic arrest of porcine oocytes. These results suggest that pWEE1B is activated after phosphorylation of the Ser77 residue, which is different from the phosphorylation site that activates mWEE1B; that pWEE1B is localized in the nucleus; and that the nuclear localization is essential for its function. Key words: Intracellular localization, Phosphorylation site, Porcine oocyte, Porcine WEE1B (J. Reprod. Dev. 57: [223][224][225][226][227][228] 2011) n mammals, ovarian oocytes are arrested at the first meiotic prophase until meiotic resumption is induced by luteinizing hormone stimulation. The maintenance of low activity of the maturation promoting factor (MPF) is a prerequisite for the maintenance of this meiotic arrest of oocytes. Phosphorylation of inhibitory phosphorylation sites in CDC2, a catalytic subunit of MPF, plays a principle role in inhibiting the MPF activity of these oocytes, and WEE1B, also called WEE2, has been identified as an oocyte-specific kinase involved in this phosphorylation in mice, humans, rhesus monkeys and pigs [1][2][3][4]. The induction of germinal vesicle break down (GVBD) by WEE1B inhibition using RNA interference or antisense oligonucleotide injection has been shown in mouse, rhesus monkey and porcine immature oocytes, in which spontaneous meiotic resumption was pharmacologically prevented [2][3][4][5]. Successful meiotic resumption was also induced in porcine growing oocytes, which could not start meiosis autonomously, by the injection of WEE1B antisense RNA [6]. These studies suggest that the function of WEE1B is required for maintenance of the meiotic arrest of...

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“…This PDE activity increase could potentially be responsible for a cGMP decrease leading to meiotic resumption as described in mouse follicles (see section 2.3). Although the presence and roles of AC3 or other AC isoforms and the GPR3 receptor have not yet been determined in porcine oocytes, an inhibitory role of www.intechopen.com Gsalpha for meiotic resumption has been demonstrated as injection of an anti-Gsalpha antibody into porcine oocytes maintained in meiotic arrest with IBMX promoted cyclin B synthesis, MPF activation and GVBD (Morikawa et al, 2007). Similarly, as in mouse and Xenopus oocytes, Wee1B, the kinase which catalyzes the inhibitory phosphorylation of CDK1, is involved in meiotic arrest of porcine oocytes (Shimaoka et al, 2009).…”

Section: Role Of Camp Phosphodiesterases and Ampkmentioning

confidence: 99%

The Control of Meiotic Arrest and Resumption in Mammalian Oocytes

Bilodeau‐Goeseels¹,

Magyara²

2012

Meiosis - Molecular Mechanisms and Cytogenetic Diversity

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Meiotic Resumption of Porcine Immature Oocytes is Prevented by Ooplasmic Gs.ALPHA. Functions

Cited by 11 publications

References 38 publications

Successful Piglet Production in a Chemically Defined System for In-vitro Production of Porcine Embryos: Dibutyryl Cyclic AMP and Epidermal Growth Factor-family Peptides Support In-vitro Maturation of Oocytes in the Absence of Gonadotropins

Successful Piglet Production in a Chemically Defined System for In-vitro Production of Porcine Embryos: Dibutyryl Cyclic AMP and Epidermal Growth Factor-family Peptides Support In-vitro Maturation of Oocytes in the Absence of Gonadotropins

Analyses of the Regulatory Mechanism of Porcine WEE1B: The Phosphorylation Sites of Porcine WEE1B and Mouse WEE1B Are Different

The Control of Meiotic Arrest and Resumption in Mammalian Oocytes

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