Nest Carbon Dioxide Masks GABA-Dependent Seizure Susceptibility in the Naked Mole-Rat

African naked mole-rats were likely the first mammals to evolve eusociality, and thus required adaptations to conserve energy and tolerate the low oxygen (O-2) and high carbon dioxide (CO2) of a densely populated fossorial nest. As hypercapnia is known to suppress neuronal activity, we studied whether naked mole-rats might demonstrate energy savings in GABAergic inhibition. Using whole-colony behavioral monitoring of captive naked mole-rats, we found a durable nest, characterized by high CO2 levels, where all colony members spent the majority of their time. Analysis of the naked mole-rat genome revealed, uniquely among mammals, a histidine point variation in the neuronal potassium-chloride cotransporter 2 (KCC2). A histidine missense substitution mutation at this locus in the human ortholog of KCC2, found previously in patients with febrile seizures and epilepsy, has been demonstrated to diminish neuronal Cl- extrusion capacity, and thus impairs GABAergic inhibition. Seizures were observed, without pharmacological intervention, in adult naked mole-rats exposed to a simulated hyperthermic surface environment, causing systemic hypocapnic alkalosis. Consistent with the diminished function of KCC2, adult naked mole-rats demonstrate a reduced efficacy of inhibition that manifests as triggering of seizures at room temperature by the GABA(A) receptor (GABA(A)R) positive allosteric modulator diazepam. These seizures are blocked in the presence of nest-like levels of CO2 and likely to be mediated through GABA(A)R activity, based on in vitro recordings. Thus, altered GABAergic inhibition adds to a growing list of adaptations in the naked mole-rat and provides a plausible proximate mechanism for nesting behavior, where a return to the colony nest restores GABA-mediated inhibition.Peer reviewe

Mutant huntingtin enhances activation of dendritic Kv4 K+ channels in striatal spiny projection neurons

Huntington\u27s disease (HD) is initially characterized by an inability to suppress unwanted movements, a deficit attributable to impaired synaptic activation of striatal indirect pathway spiny projection neurons (iSPNs). To better understand the mechanisms underlying this deficit, striatal neurons in ex vivo brain slices from mouse genetic models of HD were studied using electrophysiological, optical and biochemical approaches. Distal dendrites of iSPNs from symptomatic HD mice were hypoexcitable, a change that was attributable to increased association of dendritic Kv4 potassium channels with auxiliary KChIP subunits. This association was negatively modulated by TrkB receptor signaling. Dendritic excitability of HD iSPNs was rescued by knocking-down expression of Kv4 channels, by disrupting KChIP binding, by restoring TrkB receptor signaling or by lowering mutant-Htt (mHtt) levels with a zinc finger protein. Collectively, these studies demonstrate that mHtt induces reversible alterations in the dendritic excitability of iSPNs that could contribute to the motor symptoms of HD

Liger for Next Generation Keck AO: Filter Wheel and Pupil Design

Liger is a next-generation near-infrared imager and integral field spectrograph (IFS) for the W.M. Keck Observatory designed to take advantage of the Keck All-Sky Precision Adaptive Optics (KAPA) upgrade. Liger will operate at spectral resolving powers between R$\sim$4,000 - 10,000 over a wavelength range of 0.8-2.4$\mu$m. Liger takes advantage of a sequential imager and spectrograph design that allows for simultaneous observations between the two channels using the same filter wheel and cold pupil stop. We present the design for the filter wheels and pupil mask and their location and tolerances in the optical design. The filter mechanism is a multi-wheel design drawing from the heritage of the current Keck/OSIRIS imager single wheel design. The Liger multi-wheel configuration is designed to allow future upgrades to the number and range of filters throughout the life of the instrument. The pupil mechanism is designed to be similarly upgradeable with the option to add multiple pupil mask options. A smaller wheel mechanism allows the user to select the desired pupil mask with open slots being designed in for future upgrade capabilities. An ideal pupil would match the shape of the image formed of the primary and would track its rotation. For different pupil shapes without tracking we model the additional exposure time needed to achieve the same signal to noise of an ideal pupil and determine that a set of fixed masks of different shapes provides a mechanically simpler system with little compromise in performance.Comment: 9 pages, 7 figures, 1 tabl

Liger for Next Generation Keck Adaptive Optics: Opto-Mechanical Dewar for Imaging Camera and Slicer

Liger is a next generation adaptive optics (AO) fed integral field spectrograph (IFS) and imager for the W. M. Keck Observatory. This new instrument is being designed to take advantage of the upgraded AO system provided by Keck All-Sky Precision Adaptive-optics (KAPA). Liger will provide higher spectral resolving power (R$\sim$4,000-10,000), wider wavelength coverage ($\sim$0.8-2.4 $\mu$m), and larger fields of view than any current IFS. We present the design and analysis for a custom-made dewar chamber for characterizing the Liger opto-mechanical system. This dewar chamber is designed to test and assemble the Liger imaging camera and slicer IFS components while being adaptable for future experiments. The vacuum chamber will operate below $10^{-5}$ Torr with a cold shield that will be kept below 90 K. The dewar test chamber will be mounted to an optical vibration isolation platform and further isolated from the cryogenic and vacuum systems with bellows. The cold head and vacuums will be mounted to a custom cart that will also house the electronics and computer that interface with the experiment. This test chamber will provide an efficient means of calibrating and characterizing the Liger instrument and performing future experiments.Comment: 8 pages, 6 figure

Bose-Einstein Correlations of Three Charged Pions in Hadronic Z^0 Decays

Bose-Einstein Correlations (BEC) of three identical charged pions were studied in 4 x 10^6 hadronic Z^0 decays recorded with the OPAL detector at LEP. The genuine three-pion correlations, corrected for the Coulomb effect, were separated from the known two-pion correlations by a new subtraction procedure. A significant genuine three-pion BEC enhancement near threshold was observed having an emitter source radius of r_3 = 0.580 +/- 0.004 (stat.) +/- 0.029 (syst.) fm and a strength of \lambda_3 = 0.504 +/- 0.010 (stat.) +/- 0.041 (syst.). The Coulomb correction was found to increase the \lambda_3 value by \~9% and to reduce r_3 by ~6%. The measured \lambda_3 corresponds to a value of 0.707 +/- 0.014 (stat.) +/- 0.078 (syst.) when one takes into account the three-pion sample purity. A relation between the two-pion and the three-pion source parameters is discussed.Comment: 19 pages, LaTeX, 5 eps figures included, accepted by Eur. Phys. J.

An introduction to video methods in organizational research

Video has become a methodological tool of choice for many researchers in social science, but video methods are relatively new to the field of organization studies. This article is an introduction to video methods. First, we situate video methods relative to other kinds of research, suggesting that video recordings and analyses can be used to replace or supplement other approaches, not only observational studies but also retrospective methods such as interviews and surveys. Second, we describe and discuss various features of video data in relation to ontological assumptions that researchers may bring to their research design. Video involves both opportunities and pitfalls for researchers, who ought to use video methods in ways that are consistent with their assumptions about the world and human activity. Third, we take a critical look at video methods by reporting progress that has been made while acknowledging gaps and work that remains to be done. Our critical considerations point repeatedly at articles in this special issue, which represent recent and important advances in video method

Control and Manipulation of Pathogens with an Optical Trap for Live Cell Imaging of Intercellular Interactions

The application of live cell imaging allows direct visualization of the dynamic interactions between cells of the immune system. Some preliminary observations challenge long-held beliefs about immune responses to microorganisms; however, the lack of spatial and temporal control between the phagocytic cell and microbe has rendered focused observations into the initial interactions of host response to pathogens difficult. This paper outlines a method that advances live cell imaging by integrating a spinning disk confocal microscope with an optical trap, also known as an optical tweezer, in order to provide exquisite spatial and temporal control of pathogenic organisms and place them in proximity to host cells, as determined by the operator. Polymeric beads and live, pathogenic organisms (Candida albicans and Aspergillus fumigatus) were optically trapped using non-destructive forces and moved adjacent to living cells, which subsequently phagocytosed the trapped particle. High resolution, transmitted light and fluorescence-based movies established the ability to observe early events of phagocytosis in living cells. To demonstrate the broad applicability of this method to immunological studies, anti-CD3 polymeric beads were also trapped and manipulated to form synapses with T cells in vivo, and time-lapse imaging of synapse formation was also obtained. By providing a method to exert fine control of live pathogens with respect to immune cells, cellular interactions can be captured by fluorescence microscopy with minimal perturbation to cells and can yield powerful insight into early responses of innate and adaptive immunity.National Institute of Biomedical Imaging and Bioengineering (U.S.) (grant T32EB006348)Massachusetts General Hospital (Department of Medicine Internal Funds)Center for Computational and Integrative Biology (Development fund)Center for Computational and Integrative Biology (AI062773)Center for Computational and Integrative Biology (grant AI062773)Center for Computational and Integrative Biology (grant DK83756)Center for Computational and Integrative Biology (grant DK 043351)National Institute of Allergy and Infectious Diseases (U.S.)National Institutes of Health (U.S.) (grant AI057999

Phylogenetic Hypotheses for the Monocotyledons Constructed from rbcL Sequence Data

DNA sequences for the plastid locus that encodes the large subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase (rbcL) were determined for 18 species of monocotyledons in 15 families. These data were analyzed together with sequences for 60 other monocot species in a total of 52 families by the maximum likelihood method producing one, presumably optimal, topology. An additional 26 species were added (104 total monocot species) and analyzed by the parsimony method with an outgroup of 18 dicot species producing 109 trees of 3,932 steps. The rbcL data show at least moderate support for seven lineages corresponding to the following orders, superorders, or combinations: Arecanae; Asparagales (excluding Hypoxidaceae) plus Iridaceae; Cyclanthanae plus Pandananae; Dioscoreales; Orchidales; Typhales; and Zingiberanae. Six clades corresponding to families or genera are well supported, including: Agavaceae, Asphodelaceae, Bromeliaceae, Hypoxidaceae, Poaceae, and Tradescantia. The two, earliest diverging multispecies clades in our rbcL phylogenies, Alismatanae and Aranae, are only weakly supported, and Bromelianae, Commelinanae, and Lilianae are paraphyletic. In all analyses Acorus calamus is phylogenetically isolated as the sister species to the remaining species of monocotyledons

Genetic Evidence for Involvement of Neuronally Expressed S1P1 Receptor in Nociceptor Sensitization and Inflammatory Pain

Sphingosine-1-phosphate (S1P) is a key regulator of immune response. Immune cells, epithelia and blood cells generate high levels of S1P in inflamed tissue. However, it is not known if S1P acts on the endings of nociceptive neurons, thereby contributing to the generation of inflammatory pain. We found that the S1P1 receptor for S1P is expressed in subpopulations of sensory neurons including nociceptors. Both S1P and agonists at the S1P1 receptor induced hypersensitivity to noxious thermal stimulation in vitro and in vivo. S1P-induced hypersensitivity was strongly attenuated in mice lacking TRPV1 channels. S1P and inflammation-induced hypersensitivity was significantly reduced in mice with a conditional nociceptor-specific deletion of the S1P1 receptor. Our data show that neuronally expressed S1P1 receptors play a significant role in regulating nociceptor function and that S1P/S1P1 signaling may be a key player in the onset of thermal hypersensitivity and hyperalgesia associated with inflammation