273 research outputs found

    Cell cycle and developmental control of cortical excitability in Xenopus laevis

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    Interest in cortical excitability—the ability of the cell cortex to generate traveling waves of protein activity—has grown considerably over the past 20 years. Attributing biological functions to cortical excitability requires an understanding of the natural behavior of excitable waves and the ability to accurately quantify wave properties. Here we have investigated and quantified the onset of cortical excitability in Xenopus laevis eggs and embryos and the changes in cortical excitability throughout early development. We found that cortical excitability begins to manifest shortly after egg activation. Further, we identified a close relationship between wave properties—such as wave frequency and amplitude—and cell cycle progression as well as cell size. Finally, we identified quantitative differences between cortical excitability in the cleavage furrow relative to nonfurrow cortical excitability and showed that these wave regimes are mutually exclusive

    Rho and F-actin self-organize within an artificial cell cortex

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    The cell cortex, comprised of the plasma membrane and underlying cytoskeleton, undergoes dynamic reorganizations during a variety of essential biological processes including cell adhesion, cell migration, and cell division(1,2). During cell division and cell locomotion, for example, waves of filamentous-actin (F-actin) assembly and disassembly develop in the cell cortex in a process termed “cortical excitability”(3–7). In developing frog and starfish embryos, cortical excitability is generated through coupled positive and negative feedback, with rapid activation of Rho-mediated F-actin assembly followed in space and time by F-actin-dependent inhibition of Rho(7,8). These feedback loops are proposed to serve as a mechanism for amplification of active Rho signaling at the cell equator to support furrowing during cytokinesis, while also maintaining flexibility for rapid error correction in response to movement of the mitotic spindle during chromosome segregation(9). In this paper, we develop an artificial cortex based on Xenopus egg extract and supported lipid bilayers (SLBs), to investigate cortical Rho and F-actin dynamics(10). This reconstituted system spontaneously develops two distinct types of self-organized cortical dynamics: singular excitable Rho and F-actin waves, and non-traveling oscillatory Rho and F-actin patches. Both types of dynamic patterns have properties and dependencies similar to the excitable dynamics previously characterized in vivo(7). These findings directly support the long-standing speculation that the cell cortex is a self-organizing structure and present a novel approach for investigating mechanisms of Rho-GTPase-mediated cortical dynamics

    A versatile cortical pattern-forming circuit based on Rho, F-actin, Ect2 and RGA-3/4

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    Many cells can generate complementary traveling waves of actin filaments (F-actin) and cytoskeletal regulators. This phenomenon, termed cortical excitability, results from coupled positive and negative feedback loops of cytoskeletal regulators. The nature of these feedback loops, however, remains poorly understood. We assessed the role of the Rho GAP RGA-3/4 in the cortical excitability that accompanies cytokinesis in both frog and starfish. RGA-3/4 localizes to the cytokinetic apparatus, “chases” Rho waves in an F-actin–dependent manner, and when coexpressed with the Rho GEF Ect2, is sufficient to convert the normally quiescent, immature Xenopus oocyte cortex into a dramatically excited state. Experiments and modeling show that changing the ratio of RGA-3/4 to Ect2 produces cortical behaviors ranging from pulses to complex waves of Rho activity. We conclude that RGA-3/4, Ect2, Rho, and F-actin form the core of a versatile circuit that drives a diverse range of cortical behaviors, and we demonstrate that the immature oocyte is a powerful model for characterizing these dynamics

    The San Antonio River Mammoth Site: Archaeological Testing Investigations for the Interstate 37 Bridge at the San Antonio River Improvement Project, Bexar County, Texas

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    On behalf of the Texas Department of Transportation (TxDOT), SWCA Environmental Consultants (SWCA) conducted test excavations on the San Antonio River Mammoth site (41BX1239) and 41BX1240 and surveys in the area of potential effects (APE) of the Interstate Highway (IH) 37 bridge project at the San Antonio River in southeastern Bexar County, Texas. Work was initiated to address the requirements of Section 106 of the National Historic Preservation Act (1966) as Amended and the Antiquities Code of Texas. The purpose of the investigations was to identify, delineate, and evaluate the significance of all archaeological and historic properties potentially affected by the undertaking and, if warranted, recommend the scope of additional work. Of particular concern, site 41BX1239 contains the remains of at least two mammoths with possible evidence of cultural association based on the initial investigations by Texas A&M in 1997. However, subsequent faunal analysis, conducted by Olga Potapova and Larry D. Agenbroad of the Mammoth Site in Hot Springs, North Dakota, found inconclusive evidence for definite or valid cultural modification to the specimens studied. The testing investigations on the San Antonio River Mammoth site included the re-exposure of the original Texas A&M 1997 site trench; limited hand-excavated units to further assess the prior interpretations of the deposits and recover a sample of bone; and a detailed geomorphological assessment. The work identified a bone bed consisting of the remains of at least two mammoths. Flotation of recovered sediments from these hand excavations identified flakes of siliceous material that are consistent with micro-debitage produced by the use and retouch of stone tools. Although at the highest thresholds of certainty, the cumulative evidence is likely yet insufficient to conclusively prove human interaction with the mammoth remains, the additional data gathered herein lend some credence to the prior interpretation of the site as archaeological rather than strictly paleontological. Concurring with the previous determination, the site is considered eligible for inclusion to the National Register of Historic Places (NRHP) and for listing as a State Archeological Landmark (SAL). However, the investigations determined the site deposits are located outside the APE of the current undertaking, and therefore the project will not affect deposits associated with the San Antonio River Mammoth site. The investigations of 41BX1240 identified only a very sparse scatter of primarily surficial materials in a heavily disturbed context with no associated features or diagnostic materials. Accordingly, the site is not recommended as eligible for listing on the NRHP or for designation as a SAL. The survey identified no new archaeological sites. Based on the avoidance of 41BX1239, it is SWCA’s recommendation that no archaeological properties will be affected by the IH 37 bridge rehabilitation
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