Spontaneous Overactivation of Xenopus Frog Eggs Triggers Necrotic Cell Death.

The excessive activation of frog eggs, referred to as overactivation, can be initiated by strong oxidative stress, leading to expedited calcium-dependent non-apoptotic cell death. Overactivation also occurs spontaneously, albeit at a low frequency, in natural populations of spawned frog eggs. Currently, the cytological and biochemical events of the spontaneous process have not been characterized. In the present study, we demonstrate that the spontaneous overactivation of Xenopus frog eggs, similarly to oxidative stress- and mechanical stress-induced overactivation, is characterized by the fast and irreversible contraction of the egg's cortical layer, an increase in egg size, the depletion of intracellular ATP, a drastic increase in the intracellular ADP/ATP ratio, and the degradation of M phase-specific cyclin B2. These events manifest in eggs in the absence of caspase activation within one hour of triggering overactivation. Importantly, substantial amounts of ATP and ADP leak from the overactivated eggs, indicating that plasma membrane integrity is compromised in these cells. The rupture of the plasma membrane and acute depletion of intracellular ATP explicitly define necrotic cell death. Finally, we report that egg overactivation can occur in the frog's genital tract. Our data suggest that mechanical stress may be a key factor promoting egg overactivation during oviposition in frogs.

Polarization-insensitive local-oscillator-carrier loopback modulation for a cost-effective and high-port-count wavelength-routing optical switch.

A wavelength-routing optical switch uses a wavelength-tunable laser at each input port, and this transmitter implements output port selection by tuning the wavelength that is associated with each output port. With coherent transmission, loopback modulation of a local oscillator (LO) carrier generated at the output port can eliminate the need for a wavelength-tunable laser. However, loopback modulation can be unstable since the power fluctuates because fiber traversal by the light creates polarization rotation. Here, we propose a simple polarization-alignment circuit and verify its effectiveness in creating a high-port-count optical switch system. The proposed circuit consists of passive components and aligns the polarization state of the supplied LO carrier to be linearly polarized along the x-direction of a TE-input dual-polarization (DP) IQ modulator. The circuit is shown to yield stable modulation with Q-variation of less than 0.8 dB, regardless of any birefringence along the transmission path. The proposal's effectiveness is verified in optical switch system experiments with DP-QPSK signals; 1,856 × 1,856 switch scale is achieved with loopback modulation.

Design and verification of a LO bank enabled by fixed-wavelength lasers and fast tunable silicon ring filters for creating large scale optical switches.

The fast and widely tunable wavelength bank is a key enabler in creating wavelength-routing optical switches that do not use fast wavelength tunable lasers. A cost-effective design criterion needs to be developed before it can be applied to intra data center networks. In this paper, we develop a systematic method for designing a wavelength bank that yields high port-count and fast wavelength-routing optical switches for intra data center application. The wavelength bank is created with fixed-wavelength laser sources and wavelength-tunable filters with rapid wavelength selectivity. To optimize the optical switching system that uses the wavelength bank for supplying local oscillator (LO) lights for coherent detection, various parameters are analyzed, including effective bandwidth, laser output power, loss distribution, splitter port count, and optical amplifier gain. We carry out numerical simulations for optimizing the tradeoff between system performance and cost. To verify the designed wavelength bank, a silicon ring filter is newly fabricated with an average fiber-to-fiber insertion loss of 5.3 dB over a 22-nm bandwidth. Using 256-Gb/s DP-QPSK signals, experiments demonstrate a 1,024×1,024 optical switch that uses a fabricated silicon ring filter. The effectiveness of the scalable and fast-tunable LO bank is verified by achieving 262.1-Tb/s switch throughput with switching time under 18 µs.

Toward the understanding of biology of oocyte life cycle in Xenopus Laevis: No oocytes left behind.

BACKGROUND: For the past more than 25 years, we have been focusing on the developmental and reproductive biology of the female gametes, oocytes, and eggs, of the African clawed frog Xenopus laevis. METHODS: The events associated with the life cycle of these cells can be classified into the four main categories: first, oogenesis and cell growth in the ovary during the first meiotic arrest; second, maturation and ovulation that occur simultaneously and result in the acquisition of fertilization competence and the second meiotic arrest; third, fertilization, that is sperm-induced transition from egg to zygote; and fourth, egg death after spontaneous activation in the absence of fertilizing sperm. MAIN FINDINGS: Our studies have demonstrated that signal transduction system involving tyrosine kinase Src and other oocyte/egg membrane-associated molecules such as uroplakin III and some other cytoplasmic proteins such as mitogen-activated protein kinase (MAPK) play important roles for successful ovulation, maturation, fertilization, and initiation of embryonic development. CONCLUSION: We summarize recent advances in understanding cellular and molecular mechanisms underlying life cycle events of the oocytes and eggs. Our further intention is to discuss and predict potentially promising impact of the recent findings on the challenges facing reproductive biology and medicine, as well as societal contexts.

Autophosphorylation of MAP kinase disables the MAPK pathway in apoptotic Xenopus eggs.

Mitogen-activated protein kinases (MAPKs) are involved in the regulation of various cellular processes, including cell survival and apoptosis. Here, we report that Xenopus p42 MAPK becomes phosphorylated in apoptotic eggs, however this modification does not activate the enzyme. Using phosphorylation residue-specific antibodies, we demonstrate that this modification occurs on the Tyr residue in the MAPK activation segment, pinpointing the autophosphorylation mechanism. Notably, MAPK phosphorylation in apoptotic Xenopus eggs coincides with prominent intracellular acidification accompanying apoptosis in these cells. Furthermore, autophosphorylation of recombinant Xenopus MAPK is stimulated and phosphorylation of a protein substrate is inhibited under low pH conditions. Thus, acidic intracellular conditions inactivate MAPK and effectively disable the MAPK-mediated survival pathway in the apoptotic eggs. Given that cell acidification is a rather common feature of apoptosis, we hypothesize that stimulation of MAPK autophosphorylation and shutdown of the MAPK pathway may represent universal traits of apoptotic cell death.

Design and evaluation of quasi-Nyquist WDM networks utilizing widely deployed wavelength-selective switches.

Photonic networks based on wavelength-selective switches (WSSs) can transport wavelength-division-multiplexed (WDM) signals in a cost-effective manner. To accommodate the ever-increasing network traffic, the spectral efficiency should be maximized by minimizing the bandwidth of the guardbands inserted between WDM signals. Quasi-Nyquist WDM systems are seen as offering the highest spectral efficiency in a feasible way. However, highly dense WDM systems suffer from the signal-spectrum narrowing induced by the non-rectangular passbands of WSSs. Furthermore, widely deployed WSSs cannot process quasi-Nyquist WDM signals since the signal-alignment granularity does not match the passband resolution of the WSSs. This paper proposes a network architecture that enables quasi-Nyquist WDM networking with widely deployed WSSs. Through intensive network analyses based on computer simulations, we confirm that it has 30.8% higher spectral efficiency than conventional networks. Its feasibility is verified by transmission experiments on 72-channel 32-Gbaud/400-Gbps dual-carrier dual-polarization 16-ary quadrature-amplitude-modulation signals aligned with 66.6-GHz spacing in the full C-band. The net fiber capacity of 28.8 Tbps, the transmission distance of 900 km, and the hop count of 9 are attained by our proposed quasi-Nyquist WDM networking scheme.

In Vitro Reconstruction of Xenopus Oocyte Ovulation.

Progesterone is widely used to induce maturation of isolated fully grown oocytes of the African clawed frog, Xenopus laevis. However, the hormone fails to release oocytes from the layer of surrounding follicle cells. Here, we report that maturation and follicle rupture can be recapitulated in vitro by treating isolated follicular oocytes with progesterone and low doses of the matrix metalloproteinase (MMP), collagenase, which are ineffective in the absence of the steroid. Using this in vitro ovulation model, we demonstrate that germinal vesicle breakdown (GVBD) and oocyte liberation from ovarian follicles occur synchronously during ovulation. Inhibition of the MAPK pathway in these experimental settings suppresses both GVBD and follicular rupture, whereas inhibition of MMP activity delays follicular rupture without affecting GVBD. These results highlight importance of MAPK and MMP activities in the ovulation process and provide the first evidence for their involvement in the release of oocytes from ovarian follicles in frogs. The in vitro ovulation model developed in our study can be employed for further dissection of ovulation.

Postovulatory cell death: why eggs die via apoptosis in biological species with external fertilization.

Spawned unfertilized eggs have been found to die by apoptosis in several species with external fertilization. However, there is no necessity for the externally laid eggs to degrade via this process, as apoptosis evolved as a mechanism to reduce the damaging effects of individual cell death on the whole organism. The recent observation of egg degradation in the genital tracts of some oviparous species provides a clue as to the physiological relevance of egg apoptosis in these animals. We hypothesize that egg apoptosis accompanies ovulation in species with external fertilization as a normal process to eliminate mature eggs retained in the genital tract after ovulation. Furthermore, apoptosis universally develops in ovulated eggs after spontaneous activation in the absence of fertilization. This paper provides an overview of egg apoptosis in several oviparous biological species, including frog, fish, sea urchin, and starfish.

The egg membrane microdomain-associated uroplakin III-Src system becomes functional during oocyte maturation and is required for bidirectional gamete signaling at fertilization in Xenopus laevis.

In Xenopus laevis, sperm-egg interaction promotes partial proteolysis and/or tyrosine phosphorylation of uroplakin III (UPIII) and the tyrosine kinase Src, which both localize to the cholesterol-enriched egg membrane microdomains (MDs). Here we show that sperm promote proteolysis and/or tyrosine phosphorylation of UPIII and Src in MDs isolated from ovulated and unfertilized eggs (UF-MDs). An antibody against the extracellular domain of UPIII interferes with these events. Inhibition of fertilization by anti-UPIII antibody is rescued by co-incubation with UF-MDs. This suggests that, like MDs in intact eggs, the isolated UF-MDs are capable of interacting with sperm, an interaction that does not interfere with normal fertilization but rather augments the ability of sperm to fertilize eggs pretreated with anti-UPIII antibody. This unexpected effect of UF-MDs on sperm requires UPIII function in UF-MDs and protein kinase activity in sperm. MDs isolated from progesterone-treated mature oocytes, but not ovarian immature oocytes, are similarly functional as UF-MDs. The anti-UPIII extracellular domain antibody binds more effectively to the surface of mature than immature ovarian oocytes. We propose that the structural and functional competency of the UPIII-Src signaling system in MDs is strictly regulated during oocyte maturation and subsequently in sperm-mediated egg activation and fertilization. The fertilization-related signaling properties seen in UF-MDs can be partially reconstituted in MDs of human embryonic kidney 293 cells (293-MDs) expressing UPIII, Src and uroplakin Ib. However, 293-MDs expressing a proteolysis-resistant mutant of UPIII are less functional, suggesting that the availability of UPIII to protease action is important for MD function.

Highly reliable and large-scale subsystem-modular optical cross-connect.

To create reliable high-capacity photonic networks, we propose a novel optical cross-connect (OXC) architecture that offers failure resiliency and port scalability, simultaneously. The proposed OXC employs the subsystem-modular structure to attain the port scalability, where the use of 1xM wavelength selective switches (WSSs) or MxM WSSs is considered. Furthermore, by introducing an intra-node protection mechanism suited to each OXC architecture, our proposed scheme offers high reliability while retaining the port scalability. Through computer simulations, we evaluate the total number of WSSs needed in a network and the annual path downtime due to WSS failures. The proposed OXC architecture can drastically decrease the annual path downtime with just a small number of WSSs.

Compact OXC architecture, design and prototype development for flexible waveband routing optical networks.

A novel compact OXC node architecture that combines WSSs and arrays of small scale optical delivery-coupling type switches ("DCSWs") is proposed. Unlike conventional OXC nodes, the WSSs are only responsible for dynamic path bundling ("flexible waveband") while the small scale optical switches route bundled path groups. A network design algorithm that is aware of the routing scheme is also proposed, and numerical experiments elucidate that the necessary number of WSSs and amplifiers can be significantly reduced. A prototype of the proposed OXC is also developed using monolithic arrayed DCSWs. Transmission experiments on the prototype verify the proposal's technical feasibility.

Global decay of mRNA is a hallmark of apoptosis in aging Xenopus eggs.

Cytoplasmic mRNAs are specifically degraded in somatic cells as a part of early apoptotic response. However, no reports have been presented so far concerning mRNA fate in apoptotic gametes. In the present study, we analyzed the content of various cytoplasmic mRNAs in aging oocytes and eggs of the African clawed frog, Xenopus laevis. To circumvent large gene expression variation among the individual oocytes and eggs, single-cell monitoring of transcript levels has been implemented, using multiple cytoplasmic collections and reverse transcriptase quantitative PCR. It was found that numerous cytoplasmic mRNAs, coding for proteins classified in different functional types, are robustly degraded in apoptotic Xenopus eggs, but not in aging oocytes. mRNA degradation becomes evident in the eggs after meiotic exit at the time of cytochrome c release. A strong correlation between the length of PCR amplicon and specific transcript content was observed, suggesting endonucleolytic cleavage of mRNA. In addition, it was found that mRNA deadenylation also contributes to apoptotic mRNA degradation. Altogether, these findings indicate that the global decay of mRNA represents a hallmark of apoptosis in aging Xenopus eggs. To our knowledge, this is the first description of mRNA degradation in apoptotic gamete cells.

Highly spectral efficient networks based on grouped optical path routing.

In order to mitigate the signal spectrum narrowing caused by optical filtering at nodes, an adequate guard band is needed between optical channels, which degrades the frequency utilization of optical fibers. In this study, we propose a grouped routing based network architecture that minimizes spectrum narrowing while greatly improving spectral efficiency. Coarse granular routing at GRE (grouped routing entity) level is employed at each ROADM node, but fine granular add/drop is adopted to retain high frequency utilization. Optical channels are packed densely in each GRE, and sufficient guard bands are inserted between GREs. As a result, signal spectrum narrowing is minimized and efficient spectrum utilization is achieved. Network design/control algorithms that support both static and dynamic traffic growth are developed. Extensive simulations demonstrate the effectiveness of the proposed architecture. To implement the scheme, current LCOS-based ROADMs are applied without any hardware changes; only the control schema are modified.

Performance analysis of large-scale OXC that enables dynamic modular growth.

We investigate the performance of our recently proposed novel subsystem modular OXC node that achieves graceful modular growth without any service disruption. Simulation experiments under a realistic traffic increase scenario verify that the node architecture offers almost the same performance as the conventional large scale OXC node, while greatly reducing the number of WSSs needed and allowing the use of small degree WSSs. It is shown that both the number of WSSs traversed and total end-to-end optical loss can be substantially reduced.

Experimental verification of highly scalable OXC that consists of subsystem-modular express-switch part and multicast-switch-based add/drop part enabling total throughput of 314 Tbps.

We propose a cost-effective and scalable OXC/ROADM that consists of a subsystem-modular express switch part and a transponder-bank-based add/drop part. The effectiveness of the proposed architecture is verified via a hardware scale evaluation, network performance simulations, and transmission experiments. The architecture enables large throughput and offers significant hardware-scale reductions with marginal fiber-utilization penalty against the conventional architectures. A part of the OXC/ROADM designed to accommodate 35x35 express fiber ports and 2,800 transponders for add/drop is constructed. Its net throughput reaches 314 Tbps using 80 channels of 120-Gbps signal (30-Gbaud dual-polarization quadrature phase-shift-keying signals with 7% overhead are assumed).

[A case of atherothrombotic embolization developing with slowly progressive symptoms and requiring differential diagnosis from metastatic tumor recurrence].

We report a case of atherothrombotic embolization that developed with slowly progressive symptoms and required differential diagnosis from metastatic tumor recurrence. A 64-year-old man with a history of lung cancer and metastatic brain tumor was carefully followed at our outpatient department for tumor recurrence. Five years after surgery for brain metastasis and whole brain radiation therapy, he had no recurrence and systemic disease was well controlled. At a routine follow up in October 2013, he complained of slight right arm dysesthesia. Follow up brain magnetic resonance (MR) imaging revealed no lesion. Two months later, he developed right hemiparesthesia and gait disturbance. Spinal MR imaging was unremarkable. However, at a routine follow up in January 2014, multiple enhancements were detected near the resection cavity and regions delineating the sulci. At first, this was diagnosed as tumor recurrence. However, 3 days later, additional MR imaging detected new multiple small infarctions after worsening right hemiparesis and dysarthria. With the diagnosis of embolic stroke, we searched for an embolic source. Cardiogenic embolization and carotid bifurcation stenosis studies were negative, but severe stenosis and thrombosis were detected near the left common carotid artery origin. This site was in the field of radiation the patient received as treatment for primary lung cancer.

Performance evaluation of large-scale multi-stage hetero-granular optical cross-connects.

We proposed generalized large-scale two-stage-routing OXC architectures and evaluated the performance possible with the use of small-degree WSSs and simple optical devices; 1 x n switches or WBSSs. Numerical evaluations verify that the new architectures reduce necessary hardware scale substantially at the cost of few additional fibers while their effectiveness increases with the traffic demand. The tradeoff between the link resource increase and the hardware scale reduction is also clarified.

A novel elastic optical path network that utilizes bitrate-specific anchored frequency slot arrangement.

We propose a novel elastic optical path network where each specific bitrate signal uses its own dedicated fixed grid and one edge of its frequency grid is anchored at a specific frequency. Numerical evaluations using various bitrate signal patterns and network topologies show that the network proposal can almost match the performance of conventional flexible grid networks, while greatly mitigating the hardware requirements: it allows the use of the tunable filters for the fixed grid systems.

Transmembrane signal transduction in oocyte maturation and fertilization: focusing on Xenopus laevis as a model animal.

Fertilization is a cell biological phenomenon of crucial importance for the birth of new life in a variety of multicellular and sexual reproduction species such as algae, animal and plants. Fertilization involves a sequence of events, in which the female gamete "egg" and the male gamete "spermatozoon (sperm)" develop, acquire their functions, meet and fuse with each other, to initiate embryonic and zygotic development. Here, it will be briefly reviewed how oocyte cytoplasmic components are orchestrated to undergo hormone-induced oocyte maturation and sperm-induced activation of development. I then review how sperm-egg membrane interaction/fusion and activation of development in the fertilized egg are accomplished and regulated through egg coat- or egg plasma membrane-associated components, highlighting recent findings and future directions in the studies using Xenopus laevis as a model experimental animal.

Calcium signaling and meiotic exit at fertilization in Xenopus egg.

Calcium is a universal messenger that mediates egg activation at fertilization in all sexually reproducing species studied. However, signaling pathways leading to calcium generation and the mechanisms of calcium-induced exit from meiotic arrest vary substantially among species. Here, we review the pathways of calcium signaling and the mechanisms of meiotic exit at fertilization in the eggs of the established developmental model, African clawed frog, Xenopus laevis. We also discuss calcium involvement in the early fertilization-induced events in Xenopus egg, such as membrane depolarization, the increase in intracellular pH, cortical granule exocytosis, cortical contraction, contraction wave, cortical rotation, reformation of the nuclear envelope, sperm chromatin decondensation and sister chromatid segregation.