A retinal circuit model accounting for wide-field amacrine cells

In previous experimental studies on the visual processing in vertebrates, higher-order visual functions such as the object segregation from background were found even in the retinal stage. Previously, the “linear–nonlinear” (LN) cascade models have been applied to the retinal circuit, and succeeded to describe the input-output dynamics for certain parts of the circuit, e.g., the receptive field of the outer retinal neurons. And recently, some abstract models composed of LN cascades as the circuit elements could explain the higher-order retinal functions. However, in such a model, each class of retinal neurons is mostly omitted and thus, how those neurons play roles in the visual computations cannot be explored. Here, we present a spatio-temporal computational model of the vertebrate retina, based on the response function for each class of retinal neurons and on the anatomical inter-cellular connections. This model was capable of not only reproducing the spatio-temporal filtering properties of the outer retinal neurons, but also realizing the object segregation mechanism in the inner retinal circuit involving the “wide-field” amacrine cells. Moreover, the first-order Wiener kernels calculated for the neurons in our model showed a reasonable fit to the kernels previously measured in the real retinal neuron in situ.     

影像设备电路板测试与故障诊断之系统实现

大型医学影像设备是医院中的贵重设备,其中不少进口设备的维护修理的技术难度较高,医院每年花费在医学影像设备上的维护修理费是非常之大。本文从建立国内的维修力量着眼,研讨医学影像设备维修方法。     

G proteins and autocrine signaling differentially regulate gonadotropin subunit expression in pituitary gonadotrope

Gonadotropin-releasing hormone (GnRH) acts at gonadotropes to direct the synthesis of the gonadotropins, follicle-stimulating hormone (FSH), and luteinizing hormone (LH). The frequency of GnRH pulses determines the pattern of gonadotropin synthesis. Several hypotheses for how the gonadotrope decodes GnRH frequency to regulate gonadotropin subunit genes differentially have been proposed. However, key regulators and underlying mechanisms remain uncertain. We investigated the role of individual G proteins by perturbations using siRNA or bacterial toxins. In LβT2 gonadotrope cells, FSHβ gene induction depended predominantly on Gα(q/11), whereas LHβ expression depended on Gα(s). Specifically reducing Gα(s) signaling also disinhibited FSHβ expression, suggesting the presence of a Gα(s)-dependent signal that suppressed FSH biosynthesis. The presence of secreted factors influencing FSHβ expression levels was tested by studying the effects of conditioned media from Gα(s) knockdown and cholera toxin-treated cells on FSHβ expression. These studies and related Transwell culture experiments implicate Gα(s)-dependent secreted factors in regulating both FSHβ and LHβ gene expression. siRNA studies identify inhibinα as a Gα(s)-dependent GnRH-induced autocrine regulatory factor that contributes to feedback suppression of FSHβ expression. These results uncover differential regulation of the gonadotropin genes by Gα(q/11) and by Gα(s) and implicate autocrine and gonadotrope-gonadotrope paracrine regulatory loops in the differential induction of gonadotropin genes.     

Designing ecological corridors in a fragmented landscape: A fuzzy approach to circuit connectivity analysis

Landscape connectivity analysis is a major tool in supporting biodiversity conservation. Several methodologies have been developed to tackle it by following two main paths. The first path exploits graph approaches and models focal nodes’ connections on a resistance/conductance matrix depending on focal species’ movement potential. The second path considers geometrical pattern analyses based on the calculation of structural landscape metrics. These approaches separately investigate functional and structural features of the landscape, and may come short of a total definition if used separately. Here we propose a new scalable, modular, participative and open-source procedure based on Fuzzy logic to combine the functional and structural aspects of connectivity. We applied this method on the highly fragmented landscape of the Po Plain, focusing on its rare and endangered plain springs named fontanili. We identified an expert panel and involved it in the assignation of permeability values of land use classes with respect to the capacity of movement of animal species typical of fontanili. We concurrently performed a quantitative evaluation of the landscape fragmentation with a moving window. We found that the functional and structural evaluations were poorly correlated in the area under study (Pearson's r = −0.35, p < 0.001). We thus integrated these two non-overlapping analyses of the landscape by Fuzzy logic using thresholds and combination weights obtained from questionnaires proposed to the expert panel. The resulting index, termed Fuzzy Functionality Index (FFI), improved the level of information associated with landscape classification. By merging functional and structural aspects of the landscape, the FFI allowed us to discriminate different functional values of equally permeable parcels and vice versa. We demonstrate that FFI may act as a conductance measure in a circuit theory approach, highlighting ecological corridors between focal points of species’ distribution. We present FFI as an effective predictive index to inspect complex and non-linear landscape dynamics.     

Simultaneous real‐time assay of copper and cadmium ions by infrared photo diode electrode implanted in the muscle of live fish

The electrical circuit of an infrared photodiode electrode (IPE) was used in the simultaneous assay of copper and cadmium ions. The electrode's cyclic voltammetry (CV), chronoamperometry and square‐wave (SW) stripping voltammetric optimum conditions were examined. Results for 0–160 mg L^?1 and 50–400 μg L^?1 SW Cu(II) Cd(II), the relative standard deviation of 0.158 Cu(II), 0.077 Cd(II) (n = 15) using 20.0 mg L^?1 have been obtained at optimum conditions. The low detection limit (S/N) was attained to be at 14.71 μg L^?1(2.31 × 10^?7 mol L^?1) Cu(II) and 18.42 μg L^?1(1.63 × 10^?7 mol L^?1) Cd(II). The handmade electrode was implanted deep in the muscle of live fish and interfaced with an electrochemical workstation. Real‐time analytical application was performed on the online assay of living tissue as the specimen was moving. The methods are deemed useful in interfaced assay for physiological control, nanodiode fabrication, and in the production of laboratory on a biochip. © 2009 Wiley Periodicals, Inc. J Biochem Mol Toxicol 23:256–262, 2009; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/jbt.20287     

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents^1,2. We developed a novel nanoinjection/electrophysiology assay that allows the detection of bioactive modulatory effects of compounds on the function of a neuronal circuit that mediates the escape response in Drosophila melanogaster^3,4. Our in vivo assay, which uses the Drosophila Giant Fiber System (GFS, Figure 1) allows screening of different types of compounds, such as small molecules or peptides, and requires only minimal quantities to elicit an effect. In addition, the Drosophila GFS offers a large variety of potential molecular targets on neurons or muscles. The Giant Fibers (GFs) synapse electrically (Gap Junctions) as well as chemically (cholinergic) onto a Peripheral Synapsing Interneuron (PSI) and the Tergo Trochanteral Muscle neuron (TTMn)⁵. The PSI to DLMn (Dorsal Longitudinal Muscle neuron) connection is dependent on Dα7 nicotinic acetylcholine receptors (nAChRs)⁶. Finally, the neuromuscular junctions (NMJ) of the TTMn and the DLMn with the jump (TTM) and flight muscles (DLM) are glutamatergic^7-12. Here, we demonstrate how to inject nanoliter quantities of a compound, while obtaining electrophysiological intracellular recordings from the Giant Fiber System¹³ and how to monitor the effects of the compound on the function of this circuit. We show specificity of the assay with methyllycaconitine citrate (MLA), a nAChR antagonist, which disrupts the PSI to DLMn connection but not the GF to TTMn connection or the function of the NMJ at the jump or flight muscles.Before beginning this video it is critical that you carefully watch and become familiar with the JoVE video titled "Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster " from Augustin et al⁷, as the video presented here is intended as an expansion to this existing technique. Here we use the electrophysiological recordings method and focus in detail only on the addition of the paired nanoinjections and monitoring technique.     

Parallel-plate Flow Chamber and Continuous Flow Circuit to Evaluate Endothelial Progenitor Cells under Laminar Flow Shear Stress

The overall goal of this method is to describe a technique to subject adherent cells to laminar flow conditions and evaluate their response to well quantifiable fluid shear stresses¹.Our flow chamber design and flow circuit (Fig. 1) contains a transparent viewing region that enables testing of cell adhesion and imaging of cell morphology immediately before flow (Fig. 11A, B), at various time points during flow (Fig. 11C), and after flow (Fig. 11D). These experiments are illustrated with human umbilical cord blood-derived endothelial progenitor cells (EPCs) and porcine EPCs^2,3.This method is also applicable to other adherent cell types, e.g. smooth muscle cells (SMCs) or fibroblasts.The chamber and all parts of the circuit are easily sterilized with steam autoclaving. In contrast to other chambers, e.g. microfluidic chambers, large numbers of cells (> 1 million depending on cell size) can be recovered after the flow experiment under sterile conditions for cell culture or other experiments, e.g. DNA or RNA extraction, or immunohistochemistry (Fig. 11E), or scanning electron microscopy⁵. The shear stress can be adjusted by varying the flow rate of the perfusate, the fluid viscosity, or the channel height and width. The latter can reduce fluid volume or cell needs while ensuring that one-dimensional flow is maintained. It is not necessary to measure chamber height between experiments, since the chamber height does not depend on the use of gaskets, which greatly increases the ease of multiple experiments. Furthermore, the circuit design easily enables the collection of perfusate samples for analysis and/or quantification of metabolites secreted by cells under fluid shear stress exposure, e.g. nitric oxide (Fig. 12)⁶.     

Systems level circuit model of C. elegans undulatory locomotion: mathematical modeling and molecular genetics

To establish the relationship between locomotory behavior and dynamics of neural circuits in the nematode C. elegans we combined molecular and theoretical approaches. In particular, we quantitatively analyzed the motion of C. elegans with defective synaptic GABA and acetylcholine transmission, defective muscle calcium signaling, and defective muscles and cuticle structures, and compared the data with our systems level circuit model. The major experimental findings are: (1) anterior-to-posterior gradients of body bending flex for almost all strains both for forward and backward motion, and for neuronal mutants, also analogous weak gradients of undulatory frequency, (2) existence of some form of neuromuscular (stretch receptor) feedback, (3) invariance of neuromuscular wavelength, (4) biphasic dependence of frequency on synaptic signaling, and (5) decrease of frequency with increase of the muscle time constant. Based on (1) we hypothesize that the Central Pattern Generator (CPG) is located in the head both for forward and backward motion. Points (1) and (2) are the starting assumptions for our theoretical model, whose dynamical patterns are qualitatively insensitive to the details of the CPG design if stretch receptor feedback is sufficiently strong and slow. The model reveals that stretch receptor coupling in the body wall is critical for generation of the neuromuscular wave. Our model agrees with our behavioral data (3), (4), and (5), and with other pertinent published data, e.g., that frequency is an increasing function of muscle gap-junction coupling. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Identifying climate refugia and its potential impact on Tibetan brown bear (Ursus arctos pruinosus) in Sanjiangyuan National Park,China

Climate change has direct impacts on wildlife and future biodiversity protection efforts. Vulnerability assessment and habitat connectivity analyses are necessary for drafting effective conservation strategies for threatened species such as the Tibetan brown bear (Ursus arctos pruinosus). We used the maximum entropy (MaxEnt) model to assess the current (1950–2000) and future (2041–2060) habitat suitability by combining bioclimatic and environmental variables, and identified potential climate refugia for Tibetan brown bears in Sanjiangyuan National Park, China. Next, we selected Circuit model to simulate potential migration paths based on current and future climatically suitable habitat. Results indicate a total area of potential suitable habitat under the current climate scenario of approximately 31,649.46 km², of which 28,778.29 km² would be unsuitable by the 2050s. Potentially suitable habitat under the future climate scenario was projected to cover an area of 23,738.6 km². Climate refugia occupied 2,871.17 km², primarily in the midwestern and northeastern regions of Yangtze River Zone, as well as the northern region of Yellow River Zone. The altitude of climate refugia ranged from 4,307 to 5,524 m, with 52.93% lying at altitudes between 4,300 and 4,600 m. Refugia were mainly distributed on bare rock, alpine steppe, and alpine meadow. Corridors linking areas of potentially suitable brown bear habitat and a substantial portion of paths with low‐resistance value were distributed in climate refugia. We recommend various actions to ameliorate the impact of climate change on brown bears, such as protecting climatically suitable habitat, establishing habitat corridors, restructuring conservation areas, and strengthening monitoring efforts.