147 research outputs found

    Observations of a high‐latitude stable electron auroral emission at ∼16 MLT during a large substorm

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    Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/94792/1/jgra21045.pd

    Image informatics strategies for deciphering neuronal network connectivity

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    Brain function relies on an intricate network of highly dynamic neuronal connections that rewires dramatically under the impulse of various external cues and pathological conditions. Among the neuronal structures that show morphologi- cal plasticity are neurites, synapses, dendritic spines and even nuclei. This structural remodelling is directly connected with functional changes such as intercellular com- munication and the associated calcium-bursting behaviour. In vitro cultured neu- ronal networks are valuable models for studying these morpho-functional changes. Owing to the automation and standardisation of both image acquisition and image analysis, it has become possible to extract statistically relevant readout from such networks. Here, we focus on the current state-of-the-art in image informatics that enables quantitative microscopic interrogation of neuronal networks. We describe the major correlates of neuronal connectivity and present workflows for analysing them. Finally, we provide an outlook on the challenges that remain to be addressed, and discuss how imaging algorithms can be extended beyond in vitro imaging studies

    ANNINE-6plus, a voltage-sensitive dye with good solubility, strong membrane binding and high sensitivity

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    We present a novel voltage-sensitive hemicyanine dye ANNINE-6plus and describe its synthesis, its properties and its voltage-sensitivity in neurons. The dye ANNINE-6plus is a salt with a double positively charged chromophore and two bromide counterions. It is derived from the zwitterionic dye ANNINE-6. While ANNINE-6 is insoluble in water, ANNINE-6plus exhibits a high solubility of around 1 mM. Nonetheless, it displays a strong binding to lipid membranes. In contrast to ANNINE-6, the novel dye can be used to stain cells from aqueous solution without surfactants or organic solvents. In neuronal membranes, ANNINE-6plus exhibits the same molecular Stark effect as ANNINE-6. As a consequence, a high voltage-sensitivity is achieved with illumination and detection in the red end of the excitation and emission spectra, respectively. ANNINE-6plus will be particularly useful for sensitive optical recording of neuronal excitation when organic solvents and surfactants must be avoided as with intracellular or extracellular staining of brain tissue

    Population Dynamics and Angler Exploitation of the Unique Muskellunge Population in Shoepack Lake, Voyageurs National Park, Minnesota

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    A unique population of muskellunge Esox masquinongy inhabits Shoepack Lake in Voyageurs National Park, Minnesota. Little is known about its status, dynamics, and angler exploitation, and there is concern for the long-term viability of this population. We used intensive sampling and mark–recapture methods to quantify abundance, survival, growth, condition, age at maturity and fecundity and angler surveys to quantify angler pressure, catch rates, and exploitation. During our study, heavy rain washed out a dam constructed by beavers Castor canadensis which regulates the water level at the lake outlet, resulting in a nearly 50% reduction in surface area. We estimated a population size of 1,120 adult fish at the beginning of the study. No immediate reduction in population size was detected in response to the loss of lake area, although there was a gradual, but significant, decline in population size over the 2-year study. Adults grew less than 50 mm per year, and relative weight (W r) averaged roughly 80. Anglers were successful in catching, on average, two fish during a full day of angling, but harvest was negligible. Shoepack Lake muskellunge exhibit much slower growth rates and lower condition, but much higher densities and angler catch per unit effort (CPUE), than other muskellunge populations. The unique nature, limited distribution, and location of this population in a national park require special consideration for management. The results of this study provide the basis for assessing the long-term viability of the Shoepack Lake muskellunge population through simulations of long-term population dynamics and genetically effective population size

    Post hoc immunostaining of GABAergic neuronal subtypes following in vivo two-photon calcium imaging in mouse neocortex

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    GABAergic neurons in the neocortex are diverse with regard to morphology, physiology, and axonal targeting pattern, indicating functional specializations within the cortical microcircuitry. Little information is available, however, about functional properties of distinct subtypes of GABAergic neurons in the intact brain. Here, we combined in vivo two-photon calcium imaging in supragranular layers of the mouse neocortex with post hoc immunohistochemistry against the three calcium-binding proteins parvalbumin, calretinin, and calbindin in order to assign subtype marker profiles to neuronal activity. Following coronal sectioning of fixed brains, we matched cells in corresponding volumes of image stacks acquired in vivo and in fixed brain slices. In GAD67-GFP mice, more than 95% of the GABAergic cells could be unambiguously matched, even in large volumes comprising more than a thousand interneurons. Triple immunostaining revealed a depth-dependent distribution of interneuron subtypes with increasing abundance of PV-positive neurons with depth. Most importantly, the triple-labeling approach was compatible with previous in vivo calcium imaging following bulk loading of Oregon Green 488 BAPTA-1, which allowed us to classify spontaneous calcium transients recorded in vivo according to the neurochemically defined GABAergic subtypes. Moreover, we demonstrate that post hoc immunostaining can also be applied to wild-type mice expressing the genetically encoded calcium indicator Yellow Cameleon 3.60 in cortical neurons. Our approach is a general and flexible method to distinguish GABAergic subtypes in cell populations previously imaged in the living animal. It should thus facilitate dissecting the functional roles of these subtypes in neural circuitry

    Simulated Effects of Recruitment Variability, Exploitation, and Reduced Habitat Area on the Muskellunge Population in Shoepack Lake, Voyageurs National Park, Minnesota

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    The genetically unique population of muskellunge Esox masquinongy inhabiting Shoepack Lake in Voyageurs National Park, Minnesota, is potentially at risk for loss of genetic variability and long-term viability. Shoepack Lake has been subject to dramatic surface area changes from the construction of an outlet dam by beavers Castor canadensis and its subsequent failure. We simulated the long-term dynamics of this population in response to recruitment variation, increased exploitation, and reduced habitat area. We then estimated the effective population size of the simulated population and evaluated potential threats to long-term viability, based on which we recommend management actions to help preserve the long-term viability of the population. Simulations based on the population size and habitat area at the beginning of a companion study resulted in an effective population size that was generally above the threshold level for risk of loss of genetic variability, except when fishing mortality was increased. Simulations based on the reduced habitat area after the beaver dam failure and our assumption of a proportional reduction in population size resulted in an effective population size that was generally below the threshold level for risk of loss of genetic variability. Our results identified two potential threats to the long-term viability of the Shoepack Lake muskellunge population, reduction in habitat area and exploitation. Increased exploitation can be prevented through traditional fishery management approaches such as the adoption of no-kill, barbless hook, and limited entry regulations. Maintenance of the greatest possible habitat area and prevention of future habitat area reductions will require maintenance of the outlet dam built by beavers. Our study should enhance the long-term viability of the Shoepack Lake muskellunge population and illustrates a useful approach for other unique populations

    A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity

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    We constructed a simple and compact imaging system designed specifically for the recording of fast neuronal activity in a 3D volume. The system uses an Yb:KYW femtosecond laser we designed for use with acousto-optic deflection. An integrated two-axis acousto-optic deflector, driven by digitally synthesized signals, can target locations in three dimensions. Data acquisition and the control of scanning are performed by a LeCroy digital oscilloscope. The total cost of construction was one order of magnitude lower than that of a typical Ti:sapphire system. The entire imaging apparatus, including the laser, fits comfortably onto a small rig for electrophysiology. Despite the low cost and simplicity, the convergence of several new technologies allowed us to achieve the following capabilities: i) full-frame acquisition at video rates suitable for patch clamping; ii) random access in under ten microseconds with dwelling ability in the nominal focal plane; iii) three-dimensional random access with the ability to perform fast volume sweeps at kilohertz rates; and iv) fluorescence lifetime imaging. We demonstrate the ability to record action potentials with high temporal resolution using intracellularly loaded potentiometric dye di-2-ANEPEQ. Our design proffers easy integration with electrophysiology and promises a more widespread adoption of functional two-photon imaging as a tool for the study of neuronal activity. The software and firmware we developed is available for download at http://neurospy.org/ under an open source license

    Nanotools for Neuroscience and Brain Activity Mapping

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    Neuroscience is at a crossroads. Great effort is being invested into deciphering specific neural interactions and circuits. At the same time, there exist few general theories or principles that explain brain function. We attribute this disparity, in part, to limitations in current methodologies. Traditional neurophysiological approaches record the activities of one neuron or a few neurons at a time. Neurochemical approaches focus on single neurotransmitters. Yet, there is an increasing realization that neural circuits operate at emergent levels, where the interactions between hundreds or thousands of neurons, utilizing multiple chemical transmitters, generate functional states. Brains function at the nanoscale, so tools to study brains must ultimately operate at this scale, as well. Nanoscience and nanotechnology are poised to provide a rich toolkit of novel methods to explore brain function by enabling simultaneous measurement and manipulation of activity of thousands or even millions of neurons. We and others refer to this goal as the Brain Activity Mapping Project. In this Nano Focus, we discuss how recent developments in nanoscale analysis tools and in the design and synthesis of nanomaterials have generated optical, electrical, and chemical methods that can readily be adapted for use in neuroscience. These approaches represent exciting areas of technical development and research. Moreover, unique opportunities exist for nanoscientists, nanotechnologists, and other physical scientists and engineers to contribute to tackling the challenging problems involved in understanding the fundamentals of brain function

    The 2.5 m Telescope of the Sloan Digital Sky Survey

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    We describe the design, construction, and performance of the Sloan Digital Sky Survey Telescope located at Apache Point Observatory. The telescope is a modified two-corrector Ritchey-Chretien design which has a 2.5-m, f/2.25 primary, a 1.08-m secondary, a Gascoigne astigmatism corrector, and one of a pair of interchangeable highly aspheric correctors near the focal focal plane, one for imaging and the other for spectroscopy. The final focal ratio is f/5. The telescope is instrumented by a wide-area, multiband CCD camera and a pair of fiber-fed double spectrographs. Novel features of the telescope include: (1) A 3 degree diameter (0.65 m) focal plane that has excellent image quality and small geometrical distortions over a wide wavelength range (3000 to 10,600 Angstroms) in the imaging mode, and good image quality combined with very small lateral and longitudinal color errors in the spectroscopic mode. The unusual requirement of very low distortion is set by the demands of time-delay-and-integrate (TDI) imaging; (2) Very high precision motion to support open loop TDI observations; and (3) A unique wind baffle/enclosure construction to maximize image quality and minimize construction costs. The telescope had first light in May 1998 and began regular survey operations in 2000.Comment: 87 pages, 27 figures. AJ (in press, April 2006
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