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https://www.readbyqxmd.com/read/28930702/gcamp-expression-in-retinal-ganglion-cells-characterized-using-a-low-cost-fundus-imaging-system
#1
Yao-Chuan Chang, Steven T Walston, Robert H Chow, James D Weiland
OBJECTIVE: Virus-transduced, intracellular-calcium indicators are effective reporters of neural activity, offering the advantage of cell-specific labeling. Due to the existence of an optimal time window for the expression of calcium indicators, a suitable tool for tracking GECI expression in vivo following transduction is highly desirable. APPROACH: We developed a noninvasive imaging approach based on a custom-modified, low-cost fundus viewing system that allowed us to monitor and characterize in vivo bright-field and fluorescence images of the mouse retina...
October 2017: Journal of Neural Engineering
https://www.readbyqxmd.com/read/28921719/inflammation-alters-ampa-stimulated-calcium-responses-in-dorsal-striatal-d2-but-not-d1-spiny-projection-neurons
#2
Carissa D Winland, Nora Welsh, Alberto Sepulveda-Rodriguez, Stefano Vicini, Kathleen A Maguire-Zeiss
Neuroinflammation precedes neuronal loss in striatal neurodegenerative diseases and can be exacerbated by the release of proinflammatory molecules by microglia. These molecules can affect trafficking of AMPARs. The preferential trafficking of calcium-permeable versus impermeable AMPARs can result in disruptions of [Ca(2+) ]i and alter cellular functions. In striatal neurodegenerative diseases, changes in [Ca(2+) ]i and L-type voltage-gated calcium channels (VGCCs) have been reported. Therefore, the present study sought to determine whether a proinflammatory environment alters AMPA-stimulated [Ca(2+) ]i through calcium-permeable AMPARs and/or L-type VGCCs in dopamine-2 and dopamine-1 expressing striatal spiny projection neurons (D2 and D1 SPNs) in the dorsal striatum...
September 16, 2017: European Journal of Neuroscience
https://www.readbyqxmd.com/read/28917183/visualisation-of-an-nspef-induced-calcium-wave-using-the-genetically-encoded-calcium-indicator-gcamp-in-u87-human-glioblastoma-cells
#3
Lynn Carr, Sylvia M Bardet, Delia Arnaud-Cormos, Philippe Leveque, Rodney P O'Connor
Cytosolic, synthetic chemical calcium indicators are typically used to visualise the rapid increase in intracellular calcium ion concentration that follows nanosecond pulsed electric field (nsPEF) application. This study looks at the application of genetically encoded calcium indicators (GECIs) to investigate the spatiotemporal nature of nsPEF-induced calcium signals using fluorescent live cell imaging. Calcium responses to 44kV/cm, 10ns pulses were observed in U87-MG cells expressing either a plasma membrane targeted GECI (GCaMP5-G), or one cytosolically expressed (GCaMP6-S), and compared to the response of cells loaded with cytosolic or plasma membrane targeted chemical calcium indicators...
September 8, 2017: Bioelectrochemistry
https://www.readbyqxmd.com/read/28874729/ratiometric-matryoshka-biosensors-from-a-nested-cassette-of-green-and-orange-emitting-fluorescent-proteins
#4
Cindy Ast, Jessica Foret, Luke M Oltrogge, Roberto De Michele, Thomas J Kleist, Cheng-Hsun Ho, Wolf B Frommer
Sensitivity, dynamic and detection range as well as exclusion of expression and instrumental artifacts are critical for the quantitation of data obtained with fluorescent protein (FP)-based biosensors in vivo. Current biosensors designs are, in general, unable to simultaneously meet all these criteria. Here, we describe a generalizable platform to create dual-FP biosensors with large dynamic ranges by employing a single FP-cassette, named GO-(Green-Orange) Matryoshka. The cassette nests a stable reference FP (large Stokes shift LSSmOrange) within a reporter FP (circularly permuted green FP)...
September 5, 2017: Nature Communications
https://www.readbyqxmd.com/read/28742117/comparison-of-gcamp3-and-gcamp6f-for-studying-astrocyte-ca2-dynamics-in-the-awake-mouse-brain
#5
Liang Ye, Mateen A Haroon, Angelica Salinas, Martin Paukert
In recent years it has become increasingly clear that astrocytes play a much more active role in neural processes than the traditional view of them as supporting cells suggests. Although not electrically excitable, astrocytes exhibit diverse Ca2+ dynamics across spatial and temporal scales, more or less dependent on the animal's behavioral state. Ca2+ dynamics range from global elevations lasting multiple seconds encompassing the soma up to the finest processes, to short elevations restricted to so-called microdomains within fine processes...
2017: PloS One
https://www.readbyqxmd.com/read/28599354/a-profile-of-auditory-responsive-neurons-in-the-larval-zebrafish-brain
#6
Gilles Vanwalleghem, Lucy A Heap, Ethan K Scott
Many features of auditory processing are conserved among vertebrates, but the degree to which these pathways are established at early stages is not well explored. In this study, we have observed single cell activity throughout the brains of larval zebrafish with the goal of identifying the cellular responses, brain regions, and brain-wide pathways through which these larvae perceive and process auditory stimuli. Using GCaMP and selective plane illumination microscopy, we find strong responses to auditory tones ranging from 100 Hz to 400 Hz...
June 9, 2017: Journal of Comparative Neurology
https://www.readbyqxmd.com/read/28596596/optical-mapping-of-neuronal-activity-during-seizures-in-zebrafish
#7
L Turrini, C Fornetto, G Marchetto, M C Müllenbroich, N Tiso, A Vettori, F Resta, A Masi, G Mannaioni, F S Pavone, F Vanzi
Mapping neuronal activity during the onset and propagation of epileptic seizures can provide a better understanding of the mechanisms underlying this pathology and improve our approaches to the development of new drugs. Recently, zebrafish has become an important model for studying epilepsy both in basic research and in drug discovery. Here, we employed a transgenic line with pan-neuronal expression of the genetically-encoded calcium indicator GCaMP6s to measure neuronal activity in zebrafish larvae during seizures induced by pentylenetretrazole (PTZ)...
June 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28501905/investigation-of-pain-mechanisms-by-calcium-imaging-approaches
#8
REVIEW
Michael Anderson, Qin Zheng, Xinzhong Dong
Due to the complex circuitry and plethora of cell types involved in somatosensation, it is becoming increasingly important to be able to observe cellular activity at the population level. In addition, since cells rely on an intricate variety of extracellular factors, it is important to strive to maintain the physiological environment. Many electrophysiological techniques require the implementation of artificially-produced physiological environments and it can be difficult to assess the activity of many cells simultaneously...
May 13, 2017: Neuroscience Bulletin
https://www.readbyqxmd.com/read/28248351/extended-depth-of-field-microscopy-with-a-high-speed-deformable-mirror
#9
William J Shain, Nicholas A Vickers, Bennett B Goldberg, Thomas Bifano, Jerome Mertz
We present a wide-field fluorescence microscopy add-on that provides a fast, light-efficient extended depth-of-field (EDOF) using a deformable mirror with an update rate of 20 kHz. Out-of-focus contributions in the raw EDOF images are suppressed with a deconvolution algorithm derived directly from the microscope 3D optical transfer function. Demonstrations of the benefits of EDOF microscopy are shown with GCaMP-labeled mouse brain tissue.
March 1, 2017: Optics Letters
https://www.readbyqxmd.com/read/28235895/spike-and-neuropeptide-dependent-mechanisms-control-gnrh-neuron-nerve-terminal-ca-2-over-diverse-time-scales
#10
Karl J Iremonger, Robert Porteous, Allan E Herbison
Fast cell-to-cell communication in the brain is achieved by action potential-dependent synaptic release of neurotransmitters. The fast kinetics of transmitter release are determined by transient Ca(2+) elevations in presynaptic nerve terminals. Neuromodulators have previously been shown to regulate transmitter release by inhibiting presynaptic Ca(2+) influx. Few studies to date have demonstrated the opposite, that is, neuromodulators directly driving presynaptic Ca(2+) rises and increases in nerve terminal excitability...
March 22, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28195691/the-gcamp-r-family-of-genetically-encoded-ratiometric-calcium-indicators
#11
Jung-Hwa Cho, Carter J Swanson, Jeannie Chen, Ang Li, Lisa G Lippert, Shannon E Boye, Kasey Rose, Sivaraj Sivaramakrishnan, Cheng-Ming Chuong, Robert H Chow
We report on GCaMP-Rs, a new family of genetically encoded ratiometric calcium indicators that extend the virtues of the GCaMP proteins to ratiometric measurements. We have engineered a tandem construct of calcium-dependent GCaMP and calcium-independent mCherry fluorescent proteins. The tandem design assures that the two proteins localize in the same cellular compartment(s) and facilitates pixelwise ratiometric measurements; however, Förster resonance energy transfer (FRET) between the fluorophores reduces brightness of the sensor by up to half (depending on the GCaMP variant)...
April 21, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28160463/mapping-cortical-mesoscopic-networks-of-single-spiking-cortical-or-sub-cortical-neurons
#12
Dongsheng Xiao, Matthieu P Vanni, Catalin C Mitelut, Allen W Chan, Jeffrey M LeDue, Yicheng Xie, Andrew Cn Chen, Nicholas V Swindale, Timothy H Murphy
Understanding the basis of brain function requires knowledge of cortical operations over wide-spatial scales, but also within the context of single neurons. In vivo, wide-field GCaMP imaging and sub-cortical/cortical cellular electrophysiology were used in mice to investigate relationships between spontaneous single neuron spiking and mesoscopic cortical activity. We make use of a rich set of cortical activity motifs that are present in spontaneous activity in anesthetized and awake animals. A mesoscale spike-triggered averaging procedure allowed the identification of motifs that are preferentially linked to individual spiking neurons by employing genetically targeted indicators of neuronal activity...
February 4, 2017: ELife
https://www.readbyqxmd.com/read/27984692/probing-deep-brain-circuitry-new-advances-in-in-vivo-calcium-measurement-strategies
#13
Kasey S Girven, Dennis R Sparta
The study of neuronal ensembles in awake and behaving animals is a critical question in contemporary neuroscience research. Through the examination of calcium fluctuations, which are correlated with neuronal activity, we are able to better understand complex neural circuits. Recently, the development of technologies including two-photon microscopy, miniature microscopes, and fiber photometry has allowed us to examine calcium activity in behaving subjects over time. Visualizing changes in intracellular calcium in vivo has been accomplished utilizing GCaMP, a genetically encoded calcium indicator...
February 15, 2017: ACS Chemical Neuroscience
https://www.readbyqxmd.com/read/27974609/resting-state-hemodynamics-are-spatiotemporally-coupled-to-synchronized-and-symmetric-neural-activity-in-excitatory-neurons
#14
Ying Ma, Mohammed A Shaik, Mariel G Kozberg, Sharon H Kim, Jacob P Portes, Dmitriy Timerman, Elizabeth M C Hillman
Brain hemodynamics serve as a proxy for neural activity in a range of noninvasive neuroimaging techniques including functional magnetic resonance imaging (fMRI). In resting-state fMRI, hemodynamic fluctuations have been found to exhibit patterns of bilateral synchrony, with correlated regions inferred to have functional connectivity. However, the relationship between resting-state hemodynamics and underlying neural activity has not been well established, making the neural underpinnings of functional connectivity networks unclear...
December 27, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27922063/design-and-mechanistic-insight-into-ultrafast-calcium-indicators-for-monitoring-intracellular-calcium-dynamics
#15
Nordine Helassa, Borbala Podor, Alan Fine, Katalin Török
Calmodulin-based genetically encoded fluorescent calcium indicators (GCaMP-s) are powerful tools of imaging calcium dynamics from cells to freely moving animals. High affinity indicators with slow kinetics however distort the temporal profile of calcium transients. Here we report the development of reduced affinity ultrafast variants of GCaMP6s and GCaMP6f. We hypothesized that GCaMP-s have a common kinetic mechanism with a rate-limiting process in the interaction of the RS20 peptide and calcium-calmodulin...
December 6, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27847865/in-vivo-characterization-of-distinct-modality-specific-subsets-of-somatosensory-neurons-using-gcamp
#16
Edward C Emery, Ana P Luiz, Shafaq Sikandar, Rán Magnúsdóttir, Xinzhong Dong, John N Wood
Mechanistic insights into pain pathways are essential for a rational approach to treating this vast and increasing clinical problem. Sensory neurons that respond to tissue damage (nociceptors) may evoke pain sensations and are typically classified on the basis of action potential velocity. Electrophysiological studies have suggested that most of the C-fiber nociceptors are polymodal, responding to a variety of insults. In contrast, gene deletion studies in the sensory neurons of transgenic mice have frequently resulted in modality-specific deficits...
November 2016: Science Advances
https://www.readbyqxmd.com/read/27809317/a-microfluidics-based-method-for-measuring-neuronal-activity-in-drosophila-chemosensory-neurons
#17
Lena van Giesen, G Larisa Neagu-Maier, Jae Young Kwon, Simon G Sprecher
Monitoring neuronal responses to defined sensory stimuli is a powerful and widely used approach for understanding sensory coding in the nervous system. However, providing precise, stereotypic and reproducible cues while concomitantly recording neuronal activity remains technically challenging. Here we describe the fabrication and use of a microfluidics system that allows precise temporally restricted stimulation of Drosophila chemosensory neurons with an array of different chemical cues. The system can easily be combined with genetically encoded calcium sensors, and it can measure neuronal activity at single-cell resolution in larval sense organs and in the proboscis or leg of the adult fly...
December 2016: Nature Protocols
https://www.readbyqxmd.com/read/27725685/generation-of-transgenic-marmosets-expressing-genetically-encoded-calcium-indicators
#18
Jung Eun Park, Xian Feng Zhang, Sang-Ho Choi, Junko Okahara, Erika Sasaki, Afonso C Silva
Chronic monitoring of neuronal activity in the living brain with optical imaging techniques became feasible owing to the continued development of genetically encoded calcium indicators (GECIs). Here we report for the first time the successful generation of transgenic marmosets (Callithrix jacchus), an important nonhuman primate model in neurophysiological research, which were engineered to express the green fluorescent protein (GFP)-based family of GECIs, GCaMP, under control of either the CMV or the hSyn promoter...
October 11, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27464819/calcium-imaging-of-neuronal-activity-in-free-swimming-larval-zebrafish
#19
Akira Muto, Koichi Kawakami
Visualization of neuronal activity during animal behavior is a critical step in understanding how the brain generates behavior. In the model vertebrate zebrafish, imaging of the brain has been done mostly by using immobilized fish. Here, we describe a novel method to image neuronal activity of the larval zebrafish brain during prey capture behavior. We expressed a genetically encoded fluorescent calcium indicator, GCaMP, in the optic tectum of the midbrain using the Gal4-UAS system. Tectal activity was then imaged in unrestrained larvae during prey perception...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27441501/calcium-imaging-with-genetically-encoded-indicators-in-behaving-primates
#20
Eyal Seidemann, Yuzhi Chen, Yoon Bai, Spencer C Chen, Preeti Mehta, Bridget L Kajs, Wilson S Geisler, Boris V Zemelman
Understanding the neural basis of behaviour requires studying brain activity in behaving subjects using complementary techniques that measure neural responses at multiple spatial scales, and developing computational tools for understanding the mapping between these measurements. Here we report the first results of widefield imaging of genetically encoded calcium indicator (GCaMP6f) signals from V1 of behaving macaques. This technique provides a robust readout of visual population responses at the columnar scale over multiple mm(2) and over several months...
July 21, 2016: ELife
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