Deletion of the Na/K-ATPase α1-subunit gene (Atp1a1) does not prevent cavitation of the preimplantation mouse embryo

Barcroft, Lisa C.; Moseley, Amy E.; Lingrel, Jerry B.; Watson, Andrew J.

doi:10.1016/j.mod.2004.04.005

Cited by 52 publications

(47 citation statements)

References 66 publications

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“…Embryonic Na/K-ATPase activity increases when a nascent blastocoele cavity forms and then decreases to the baseline level at full expansion [53]. Mouse embryos with targeted disruption in the Na/k-ATPase α1-subunit progress through blastulation, but they do not expand and eventually die [54]. We found lower ATP1A1 expression in fully expanded blastocysts than in blastocysts that hatched [55].…”

mentioning

confidence: 61%

Gene Expression in Early Expanded Parthenogenetic and In Vitro Fertilized Bovine Blastocysts

Gómez

Caamaño

Bermejo-Álvarez³

et al. 2009

Journal of Reproduction and Development

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Abstract. Mammalian oocytes can undergo artificial parthenogenesis in vitro and develop to the blastocyst stage. In this study, using real-time PCR, we analyzed the expression of genes representative of essential events in development. In vitro matured oocytes were either fertilized or activated with ionomycin + 6-DMAP and cultured in simple medium. The pluripotency-related gene Oct3/4 was downregulated in parthenotes, while the de novo methylation DNMT3A gene was unchanged. Among the pregnancy recognition genes, IFN-t was upregulated, PGRMC1 was downregulated and PLAC8 was unchanged in parthenotes. Among the metabolism genes, SLC2A1 was downregulated, while AKR1B1, COX2, H6PD and TXN were upregulated in parthenotes; there was no difference in SLC2A5. Among the genes involved in compaction/blastulation, GJA1 expression increased in parthenotes, but no differences were detected within ATP1A1 and CDH1. Expression of p66 shc and the Bax/Bcl2 ratio were higher in parthenotes, and there was no difference in p53. Parthenotes and embryos may differ in the way they stimulate apoptosis, with a preponderant role for p66 shc within parthenotes. Differentially affected functions may also include pluripotency, de novo methylation and early embryonic signalling. Key words: Bovine, Embryo, Gene expression, Parthenote (J. Reprod. Dev. 55: [607][608][609][610][611][612][613][614] 2009) arthenogenesis, a form of reproduction not spontaneous in mammals, is a process by which the oocyte develops without the male gamete. Mammalian parthenotes are unable to develop to term in utero (developing for a maximum of 48 days in the cow; [1]), and this is thought to be due to alterations in their genomic imprinting, a mechanism that provides complementary, but not equivalent, specific expression between maternal and paternal genomes [2]. During fertilization, the spermatozoon triggers multiple and rhythmic oscillations of intracellular free calcium that release the oocyte from this blocked stage [3]. These intracellular calcium patterns can be mimicked by a number of chemicals that activate the oocyte without sperm, yielding parthenogenetic embryos [4]. In cattle, production of reconstructed embryos by somatic cell nuclear transfer (SCNT) [5] and intracytoplasmic sperm injection (ICSI) [6] depends on successful oocyte activation. Therefore, procedures involving SCNT and ICSI could benefit from a better knowledge of mechanisms that differ between embryos and parthenotes.Contrary to the research in mice and humans, most studies have failed to detect putatively imprinted genes in bovine early development [7,8], suggesting that monoallelic expression may not develop until after Day 21 of development, as occurs in ovine species [9]. In fact, a recent study found imprinted expression of bovine Mest-1 in Day-21 but not Day-14 embryos, while 7 other putatively imprinted genes were shown to be not imprinted at the two stages analyzed [10]. However, gene expression does differ between blastocysts from embryos and parthenotes in domestic species [11...

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confidence: 61%

Gene Expression in Early Expanded Parthenogenetic and In Vitro Fertilized Bovine Blastocysts

Gómez

Caamaño

Bermejo-Álvarez³

et al. 2009

Journal of Reproduction and Development

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“…Failure of a null mutation to disrupt events in preimplantation development can mean that the gene has functions that become critical only later in development (46). Indeed, even targeted disruption of the Atp1a1 gene encoding the ␣1 subunit of Na,KATPase failed to impair blastocyst development, although the mutant embryos were eventually lost during the peri-implantation period (47).…”

Section: Discussionmentioning

confidence: 99%

Na,K-ATPase from Mice Lacking the γ Subunit (FXYD2) Exhibits Altered Na+ Affinity and Decreased Thermal Stability

Jones

Arystarkhova

et al. 2005

Journal of Biological Chemistry

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The ␥ subunit of the Na,K-ATPase, a 7-kDa single-span membrane protein, is a member of the FXYD gene family. Several FXYD proteins have been shown to bind to Na,K-ATPase and modulate its properties, and each FXYD protein appears to alter enzyme kinetics differently. Different results have sometimes been obtained with different experimental systems, however. To test for effects of ␥ in a native tissue environment, mice lacking a functional ␥ subunit gene (Fxyd2) were generated. These mice were viable and without observable pathology. Prior work in the mouse embryo showed that ␥ is expressed at the blastocyst stage. However, there was no delay in blastocele formation, and the expected Mendelian ratios of offspring were obtained even with Fxyd2 ؊/؊ dams. In adult Fxyd2 ؊/؊ mouse kidney, splice variants of ␥ that have different nephron segment-specific expression patterns were absent. Purified ␥-deficient renal Na,K-ATPase displayed higher apparent affinity for Na ؉ without significant change in apparent affinity for K ؉ . Affinity for ATP, which was expected to be decreased, was instead slightly increased. The results suggest that regulation of Na ؉ sensitivity is a major functional role for this protein, whereas regulation of ATP affinity may be context-specific. Most importantly, this implies that ␥ and other FXYD proteins have their effects by local and not global conformation change. Na,K-ATPase lacking the ␥ subunit had increased thermal lability. Combined with other evidence that ␥ participates in an early step of thermal denaturation, this indicates that FXYD proteins may play an important structural role in the enzyme complex.

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“…In the mouse, the expression of multiple isoforms of a and b subunits by the blastocyst maintains cavitation, but targeted disruption in the Na/k-ATPase a1-subunit suppresses expansion and the embryo dies (Barcroft et al 2004). Embryonic Na/K-ATPase activity increases when a nascent blastocoele cavity forms and decreases to baseline levels at full expansion (Houghton et al 2003).…”

Section: Development and Gene Expression In Parthenotesmentioning

confidence: 99%

Biological differences between in vitro produced bovine embryos and parthenotes

Gómez¹,

Gutiérrez‐Adán²,

Díez³

et al. 2009

Reproduction

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Parthenotes may represent an alternate ethical source of stem cells, once biological differences between parthenotes and embryos can be understood. In this study, we analyzed development, trophectoderm (TE) differentiation, apoptosis/necrosis, and ploidy in parthenotes and in vitro produced bovine embryos. Subsequently, using real-time PCR, we analyzed the expression of genes expected to underlie the observed differences at the blastocyst stage. In vitro matured oocytes were either fertilized or activated with ionomycin C6-DMAP and cultured in simple medium. Parthenotes showed enhanced blastocyst development and diploidy and reduced TE cell counts. Apoptotic and necrotic indexes did not vary, but parthenotes evidenced a higher relative proportion of apoptotic cells between inner cell mass and TE. The pluripotence-related POU5F1 and the methylation DNMT3A genes were downregulated in parthenotes. Among pregnancy recognition genes, TP-1 was upregulated in parthenotes, while PGRMC1 and PLAC8 did not change. Expression of p66 shc and BAX/BCL2 ratio were higher, and p53 lower, in parthenotes. Among metabolism genes, SLC2A1 was downregulated, while AKR1B1, PTGS2, H6PD, and TXN were upregulated in parthenotes, and SLC2A5 did not differ. Among genes involved in compaction/blastulation, GJA1 was downregulated in parthenotes, but no differences were detected within ATP1A1 and CDH1. Within parthenotes, the expression levels of SLC2A1, TP-1, and H6PD, and possibly AKR1B1, resemble patterns described in female embryos. The pro-apoptotic profile is more pronounced in parthenotes than in embryos, which may differ in their way to channel apoptotic stimuli, through p66 shc and p53respectively, and in their mechanisms to control pluripotency and de novo methylation.Reproduction (2009) 137 285-295

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Deletion of the Na/K-ATPase α1-subunit gene (Atp1a1) does not prevent cavitation of the preimplantation mouse embryo

Cited by 52 publications

References 66 publications

Gene Expression in Early Expanded Parthenogenetic and In Vitro Fertilized Bovine Blastocysts

Gene Expression in Early Expanded Parthenogenetic and In Vitro Fertilized Bovine Blastocysts

Na,K-ATPase from Mice Lacking the γ Subunit (FXYD2) Exhibits Altered Na+ Affinity and Decreased Thermal Stability

Biological differences between in vitro produced bovine embryos and parthenotes

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