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Artículos de revistas sobre el tema "GCaMP"

Autor: Grafiati

Publicado: 3 de julio de 2021

Última modificación: 10 de marzo de 2023

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1

Shigetomi, Eiji, Sebastian Kracun y Baljit S. Khakh. "Monitoring astrocyte calcium microdomains with improved membrane targeted GCaMP reporters". Neuron Glia Biology 6, n.º 3 (agosto de 2010): 183–91. http://dx.doi.org/10.1017/s1740925x10000219.

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Astrocytes are involved in synaptic and cerebrovascular regulation in the brain. These functions are regulated by intracellular calcium signalling that is thought to reflect a form of astrocyte excitability. In a recent study, we reported modification of the genetically encoded calcium indicator (GECI) GCaMP2 with a membrane-tethering domain, Lck, to generate Lck-GCaMP2. This GECI allowed us to detect novel microdomain calcium signals. The microdomains were random and ‘spotty’ in nature. In order to detect such signals more reliably, in the present study we further modified Lck-GCaMP2 to carry three mutations in the GCaMP2 moiety (M153K, T203V within EGFP and N60D in the CaM domain) to generate Lck-GCaMP3. We directly compared Lck-GCaMP2 and Lck-GCaMP3 by assessing their ability to monitor several types of astrocyte calcium signals with a focus on spotty microdomains. Our data show that Lck-GCaMP3 is between two- and four-times better than Lck-GCaMP2 in terms of its basal fluorescence intensity, signal-to-noise and its ability to detect microdomains. The use of Lck-GCaMP3 thus represents a significantly improved way to monitor astrocyte calcium signals, including microdomains, and will facilitate detailed exploration of their molecular mechanisms and physiological roles.

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2

Chen, Yen Lin, Thomas M. Baker, Frank Lee, Bo Shui, Jane C. Lee, Petr Tvrdik, Michael I. Kotlikoff y Swapnil K. Sonkusare. "Calcium Signal Profiles in Vascular Endothelium from Cdh5-GCaMP8 and Cx40-GCaMP2 Mice". Journal of Vascular Research 58, n.º 3 (2021): 159–71. http://dx.doi.org/10.1159/000514210.

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<b><i>Introduction:</i></b> Studies in Cx40-GCaMP2 mice, which express calcium biosensor GCaMP2 in the endothelium under connexin 40 promoter, have identified the unique properties of endothelial calcium signals. However, Cx40-GCaMP2 mouse is associated with a narrow dynamic range and lack of signal in the venous endothelium. Recent studies have proposed many GCaMPs (GCaMP5/6/7/8) with improved properties although their performance in endothelium-specific calcium studies is not known. <b><i>Methods:</i></b> We characterized a newly developed mouse line that constitutively expresses GCaMP8 in the endothelium under the VE-cadherin (Cdh5-GCaMP8) promoter. Calcium signals through endothelial IP3 receptors and TRP vanilloid 4 (TRPV4) ion channels were recorded in mesenteric arteries (MAs) and veins from Cdh5-GCaMP8 and Cx40-GCaMP2 mice. <b><i>Results:</i></b> Cdh5-GCaMP8 mice showed lower baseline fluorescence intensity, higher dynamic range, and higher amplitudes of individual calcium signals than Cx40-GCaMP2 mice. Importantly, Cdh5-GCaMP8 mice enabled the first recordings of discrete calcium signals in the intact venous endothelium and revealed striking differences in IP3 receptor and TRPV4 channel calcium signals between MAs and mesenteric veins. <b><i>Conclusion:</i></b> Our findings suggest that Cdh5-GCaMP8 mice represent significant improvements in dynamic range, sensitivity for low-intensity signals, and the ability to record calcium signals in venous endothelium.

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3

Krogman, William, J. Alan Sparks y Elison B. Blancaflor. "Cell Type-Specific Imaging of Calcium Signaling in Arabidopsis thaliana Seedling Roots Using GCaMP3". International Journal of Molecular Sciences 21, n.º 17 (2 de septiembre de 2020): 6385. http://dx.doi.org/10.3390/ijms21176385.

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Cytoplasmic calcium ([Ca2+]cyt) is a well-characterized second messenger in eukaryotic cells. An elevation in [Ca2+]cyt levels is one of the earliest responses in plant cells after exposure to a range of environmental stimuli. Advances in understanding the role of [Ca2+]cyt in plant development has been facilitated by the use of genetically-encoded reporters such as GCaMP. Most of these studies have relied on promoters such as Cauliflower Mosaic Virus (35S) and Ubiquitin10 (UBQ10) to drive expression of GCaMP in all cell/tissue types. Plant organs such as roots consist of various cell types that likely exhibit unique [Ca2+]cyt responses to exogenous and endogenous signals. However, few studies have addressed this question. Here, we introduce a set of Arabidopsis thaliana lines expressing GCaMP3 in five root cell types including the columella, endodermis, cortex, epidermis, and trichoblasts. We found similarities and differences in the [Ca2+]cyt signature among these root cell types when exposed to adenosine tri-phosphate (ATP), glutamate, aluminum, and salt, which are known to trigger [Ca2+]cyt increases in root cells. These cell type-targeted GCaMP3 lines provide a new resource that should enable more in depth studies that address how a particular environmental stimulus is linked to specific root developmental pathways via [Ca2+]cyt.

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4

Ma, Ying, Mohammed A. Shaik, Mariel G. Kozberg, Sharon H. Kim, Jacob P. Portes, Dmitriy Timerman y Elizabeth M. C. Hillman. "Resting-state hemodynamics are spatiotemporally coupled to synchronized and symmetric neural activity in excitatory neurons". Proceedings of the National Academy of Sciences 113, n.º 52 (14 de diciembre de 2016): E8463—E8471. http://dx.doi.org/10.1073/pnas.1525369113.

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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. In this study, neural activity and hemodynamics were recorded simultaneously over the bilateral cortex of awake and anesthetized Thy1-GCaMP mice using wide-field optical mapping. Neural activity was visualized via selective expression of the calcium-sensitive fluorophore GCaMP in layer 2/3 and 5 excitatory neurons. Characteristic patterns of resting-state hemodynamics were accompanied by more rapidly changing bilateral patterns of resting-state neural activity. Spatiotemporal hemodynamics could be modeled by convolving this neural activity with hemodynamic response functions derived through both deconvolution and gamma-variate fitting. Simultaneous imaging and electrophysiology confirmed that Thy1-GCaMP signals are well-predicted by multiunit activity. Neurovascular coupling between resting-state neural activity and hemodynamics was robust and fast in awake animals, whereas coupling in urethane-anesthetized animals was slower, and in some cases included lower-frequency (<0.04 Hz) hemodynamic fluctuations that were not well-predicted by local Thy1-GCaMP recordings. These results support that resting-state hemodynamics in the awake and anesthetized brain are coupled to underlying patterns of excitatory neural activity. The patterns of bilaterally-symmetric spontaneous neural activity revealed by wide-field Thy1-GCaMP imaging may depict the neural foundation of functional connectivity networks detected in resting-state fMRI.

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5

Ai, Minrong, Holly Mills, Makoto Kanai, Jason Lai, Jingjing Deng, Eric Schreiter, Loren Looger, Thomas Neubert y Greg Suh. "Green-to-Red Photoconversion of GCaMP". PLOS ONE 10, n.º 9 (18 de septiembre de 2015): e0138127. http://dx.doi.org/10.1371/journal.pone.0138127.

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6

Ivashkina, Olga I., Anna M. Gruzdeva, Marina A. Roshchina, Ksenia A. Toropova y Konstantin V. Anokhin. "Imaging of C-fos Activity in Neurons of the Mouse Parietal Association Cortex during Acquisition and Retrieval of Associative Fear Memory". International Journal of Molecular Sciences 22, n.º 15 (31 de julio de 2021): 8244. http://dx.doi.org/10.3390/ijms22158244.

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The parietal cortex of rodents participates in sensory and spatial processing, movement planning, and decision-making, but much less is known about its role in associative learning and memory formation. The present study aims to examine the involvement of the parietal association cortex (PtA) in associative fear memory acquisition and retrieval in mice. Using ex vivo c-Fos immunohistochemical mapping and in vivo Fos-EGFP two-photon imaging, we show that PtA neurons were specifically activated both during acquisition and retrieval of cued fear memory. Fos immunohistochemistry revealed specific activation of the PtA neurons during retrieval of the 1-day-old fear memory. In vivo two-photon Fos-EGFP imaging confirmed this result and in addition detected specific c-Fos responses of the PtA neurons during acquisition of cued fear memory. To allow a more detailed study of the long-term activity of such PtA engram neurons, we generated a Fos-Cre-GCaMP transgenic mouse line that employs the Targeted Recombination in Active Populations (TRAP) technique to detect calcium events specifically in cells that were Fos-active during conditioning. We show that gradual accumulation of GCaMP3 in the PtA neurons of Fos-Cre-GCaMP mice peaks at the 4th day after fear learning. We also describe calcium transients in the cell bodies and dendrites of the TRAPed neurons. This provides a proof-of-principle for TRAP-based calcium imaging of PtA functions during memory processes as well as in experimental models of fear- and anxiety-related psychiatric disorders and their specific therapies.

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7

Han, Su Young, Jenny Clarkson, Richard Piet y Allan E. Herbison. "Optical Approaches for Interrogating Neural Circuits Controlling Hormone Secretion". Endocrinology 159, n.º 11 (9 de octubre de 2018): 3822–33. http://dx.doi.org/10.1210/en.2018-00594.

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Abstract Developments in optical imaging and optogenetics are transforming the functional investigation of neuronal networks throughout the brain. Recent studies in the neuroendocrine field have used genetic mouse models combined with a variety of light-activated optical tools as well as GCaMP calcium imaging to interrogate the neural circuitry controlling hormone secretion. The present review highlights the benefits and caveats of these approaches for undertaking both acute brain slice and functional studies in vivo. We focus on the use of channelrhodopsin and the inhibitory optogenetic tools, archaerhodopsin and halorhodopsin, in addition to GCaMP imaging of individual cells in vitro and neural populations in vivo using fiber photometry. We also address issues around the use of genetic vs viral delivery of encoded proteins to specific Cre-expressing cell populations, their quantification, and the use of conscious vs anesthetized animal models. To date, optogenetics and GCaMP imaging have proven useful in dissecting functional circuitry within the brain and are likely to become essential investigative tools for deciphering the different neural networks controlling hormone secretion.

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8

Chen, Qian, Joseph Cichon, Wenting Wang, Li Qiu, Seok-Jin R. Lee, Nolan R. Campbell, Nicholas DeStefino et al. "Imaging Neural Activity Using Thy1-GCaMP Transgenic Mice". Neuron 76, n.º 2 (octubre de 2012): 297–308. http://dx.doi.org/10.1016/j.neuron.2012.07.011.

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9

Creamer, Matthew S., Kevin S. Chen, Andrew M. Leifer y Jonathan W. Pillow. "Correcting motion induced fluorescence artifacts in two-channel neural imaging". PLOS Computational Biology 18, n.º 9 (28 de septiembre de 2022): e1010421. http://dx.doi.org/10.1371/journal.pcbi.1010421.

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Imaging neural activity in a behaving animal presents unique challenges in part because motion from an animal’s movement creates artifacts in fluorescence intensity time-series that are difficult to distinguish from neural signals of interest. One approach to mitigating these artifacts is to image two channels simultaneously: one that captures an activity-dependent fluorophore, such as GCaMP, and another that captures an activity-independent fluorophore such as RFP. Because the activity-independent channel contains the same motion artifacts as the activity-dependent channel, but no neural signals, the two together can be used to identify and remove the artifacts. However, existing approaches for this correction, such as taking the ratio of the two channels, do not account for channel-independent noise in the measured fluorescence. Here, we present Two-channel Motion Artifact Correction (TMAC), a method which seeks to remove artifacts by specifying a generative model of the two channel fluorescence that incorporates motion artifact, neural activity, and noise. We use Bayesian inference to infer latent neural activity under this model, thus reducing the motion artifact present in the measured fluorescence traces. We further present a novel method for evaluating ground-truth performance of motion correction algorithms by comparing the decodability of behavior from two types of neural recordings; a recording that had both an activity-dependent fluorophore and an activity-independent fluorophore (GCaMP and RFP) and a recording where both fluorophores were activity-independent (GFP and RFP). A successful motion correction method should decode behavior from the first type of recording, but not the second. We use this metric to systematically compare five models for removing motion artifacts from fluorescent time traces. We decode locomotion from a GCaMP expressing animal 20x more accurately on average than from control when using TMAC inferred activity and outperforms all other methods of motion correction tested, the best of which were ~8x more accurate than control.

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10

Cho, Jung-Hwa, Carter J. Swanson, Jeannie Chen, Ang Li, Lisa G. Lippert, Shannon E. Boye, Kasey Rose, Sivaraj Sivaramakrishnan, Cheng-Ming Chuong y Robert H. Chow. "The GCaMP-R Family of Genetically Encoded Ratiometric Calcium Indicators". ACS Chemical Biology 12, n.º 4 (marzo de 2017): 1066–74. http://dx.doi.org/10.1021/acschembio.6b00883.

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11

Muto, Akira, Junichi Nakai y Koichi Kawakami. "Calcium imaging of the zebrafish visual system with the GCaMP". Neuroscience Research 71 (septiembre de 2011): e68-e69. http://dx.doi.org/10.1016/j.neures.2011.07.292.

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12

Akerboom, J., T. W. Chen, T. J. Wardill, L. Tian, J. S. Marvin, S. Mutlu, N. C. Calderon et al. "Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging". Journal of Neuroscience 32, n.º 40 (3 de octubre de 2012): 13819–40. http://dx.doi.org/10.1523/jneurosci.2601-12.2012.

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13

Yang, Yaxiong, Yuanyuan He y Xiaodong Liu. "Design and Applications of the New Calcium Sensor GCaMP-X". Biophysical Journal 116, n.º 3 (febrero de 2019): 111a. http://dx.doi.org/10.1016/j.bpj.2018.11.630.

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14

Mao, Tianyi, Daniel H. O'Connor, Volker Scheuss, Junichi Nakai y Karel Svoboda. "Characterization and Subcellular Targeting of GCaMP-Type Genetically-Encoded Calcium Indicators". PLoS ONE 3, n.º 3 (19 de marzo de 2008): e1796. http://dx.doi.org/10.1371/journal.pone.0001796.

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15

Weitz, Andrew C., Matthew R. Behrend, Nan Sook Lee, Ronald L. Klein, Vince A. Chiodo, William W. Hauswirth, Mark S. Humayun, James D. Weiland y Robert H. Chow. "Imaging the response of the retina to electrical stimulation with genetically encoded calcium indicators". Journal of Neurophysiology 109, n.º 7 (1 de abril de 2013): 1979–88. http://dx.doi.org/10.1152/jn.00852.2012.

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Epiretinal implants for the blind are designed to stimulate surviving retinal neurons, thus bypassing the diseased photoreceptor layer. Single-unit or multielectrode recordings from isolated animal retina are commonly used to inform the design of these implants. However, such electrical recordings provide limited information about the spatial patterns of retinal activation. Calcium imaging overcomes this limitation, as imaging enables high spatial resolution mapping of retinal ganglion cell (RGC) activity as well as simultaneous recording from hundreds of RGCs. Prior experiments in amphibian retina have demonstrated proof of principle, yet experiments in mammalian retina have been hindered by the inability to load calcium indicators into mature mammalian RGCs. Here, we report a method for labeling the majority of ganglion cells in adult rat retina with genetically encoded calcium indicators, specifically GCaMP3 and GCaMP5G. Intravitreal injection of an adeno-associated viral vector targets ∼85% of ganglion cells with high specificity. Because of the large fluorescence signals provided by the GCaMP sensors, we can now for the first time visualize the response of the retina to electrical stimulation in real-time. Imaging transduced retinas mounted on multielectrode arrays reveals how stimulus pulse shape can dramatically affect the spatial extent of RGC activation, which has clear implications in prosthetic applications. Our method can be easily adapted to work with other fluorescent indicator proteins in both wild-type and transgenic mammals.

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16

Valley, M. T., M. G. Moore, J. Zhuang, N. Mesa, D. Castelli, D. Sullivan, M. Reimers y J. Waters. "Separation of hemodynamic signals from GCaMP fluorescence measured with wide-field imaging". Journal of Neurophysiology 123, n.º 1 (1 de enero de 2020): 356–66. http://dx.doi.org/10.1152/jn.00304.2019.

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Wide-field calcium imaging is often used to measure brain dynamics in behaving mice. With a large field of view and a high sampling rate, wide-field imaging can monitor activity from several distant cortical areas simultaneously, revealing cortical interactions. Interpretation of wide-field images is complicated, however, by the absorption of light by hemoglobin, which can substantially affect the measured fluorescence. One approach to separating hemodynamics and calcium signals is to use multiwavelength backscatter recordings to measure light absorption by hemoglobin. Following this approach, we develop a spatially detailed regression-based method to estimate hemodynamics. This Spatial Model is based on a linear form of the Beer–Lambert relationship but is fit at every pixel in the image and does not rely on the estimation of physical parameters. In awake mice of three transgenic lines, the Spatial Model offers improved separation of hemodynamics and changes in GCaMP fluorescence. The improvement is pronounced near blood vessels and, in contrast with the Beer–Lambert equations, can remove vascular artifacts along the sagittal midline and in general permits more accurate fluorescence-based determination of neuronal activity across the cortex. NEW & NOTEWORTHY This paper addresses a well-known and strong source of contamination in wide-field calcium-imaging data: hemodynamics. To guide researchers toward the best method to separate calcium signals from hemodynamics, we compare the performance of several methods in three commonly used mouse lines and present a novel regression model that outperforms the other techniques we consider.

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17

Singh, Mahendra, Brendan Lujan y Robert Renden. "Presynaptic GCaMP expression decreases vesicle release probability at the calyx of Held". Synapse 72, n.º 12 (17 de julio de 2018): e22040. http://dx.doi.org/10.1002/syn.22040.

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18

Tian, Lin, S. Andrew Hires, Tianyi Mao, Daniel Huber, M. Eugenia Chiappe, Sreekanth H. Chalasani, Leopoldo Petreanu et al. "Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators". Nature Methods 6, n.º 12 (8 de noviembre de 2009): 875–81. http://dx.doi.org/10.1038/nmeth.1398.

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19

Han, Su Young, Grace Kane, Isaiah Cheong y Allan E. Herbison. "Characterization of GnRH Pulse Generator Activity in Male Mice Using GCaMP Fiber Photometry". Endocrinology 160, n.º 3 (15 de enero de 2019): 557–67. http://dx.doi.org/10.1210/en.2018-01047.

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20

Scott, Benjamin B., Stephan Y. Thiberge, Caiying Guo, D. Gowanlock R. Tervo, Carlos D. Brody, Alla Y. Karpova y David W. Tank. "Imaging Cortical Dynamics in GCaMP Transgenic Rats with a Head-Mounted Widefield Macroscope". Neuron 100, n.º 5 (diciembre de 2018): 1045–58. http://dx.doi.org/10.1016/j.neuron.2018.09.050.

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21

Emery, Edward C., Ana P. Luiz, Shafaq Sikandar, Rán Magnúsdóttir, Xinzhong Dong y John N. Wood. "In vivo characterization of distinct modality-specific subsets of somatosensory neurons using GCaMP". Science Advances 2, n.º 11 (noviembre de 2016): e1600990. http://dx.doi.org/10.1126/sciadv.1600990.

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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. We have used an in vivo imaging approach using the genetically encoded fluorescent calcium indicator GCaMP to study the activity of dorsal root ganglion sensory neurons in live animals challenged with painful stimuli. Using this approach, we can visualize spatially distinct neuronal responses and find that >85% of responsive dorsal root ganglion neurons are modality-specific, responding to either noxious mechanical, cold, or heat stimuli. These observations are mirrored in behavioral studies of transgenic mice. For example, deleting sodium channel Nav1.8 silences mechanical- but not heat-sensing sensory neurons, consistent with behavioral deficits. In contrast, primary cultures of axotomized sensory neurons show high levels of polymodality. After intraplantar treatment with prostaglandin E2, neurons in vivo respond more intensely to noxious thermal and mechanical stimuli, and additional neurons (silent nociceptors) are unmasked. Together, these studies define polymodality as an infrequent feature of nociceptive neurons in normal animals.

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22

Hartung, Jane E. y Michael S. Gold. "GCaMP as an indirect measure of electrical activity in rat trigeminal ganglion neurons". Cell Calcium 89 (julio de 2020): 102225. http://dx.doi.org/10.1016/j.ceca.2020.102225.

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23

Herdman, Ashley, Alex Lagasse, Kenzie MacNicol, Anessa Haney, Uli Boehm, Melanie MacNicol, Angus MacNicol, Gwen Childs, James Hyde y Angela Odle. "PMON55 Calcium Imaging in the Intact Mouse Pituitary: A Novel Design for Understanding the Formation and Modulation of the Gonadotrope Network". Journal of the Endocrine Society 6, Supplement_1 (1 de noviembre de 2022): A556. http://dx.doi.org/10.1210/jendso/bvac150.1155.

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Abstract The reproductive cells of the pituitary, the gonadotropes, develop after e12.75 in mice and continue to differentiate and migrate throughout puberty. In response to physiological signals, these cells change in morphology to alter connections with other cells. Previous studies have indicated that gonadotropes have functional relationships in an interconnected, organized network. However, little work has been done to determine (1) what developmental cues are needed to organize this network, or (2) how this network functions to promote gonadotropin secretion. Activation of gonadotropin-releasing hormone receptors (GnRHRs) leads to an increase in intracellular calcium; therefore, this pathway can be used for visualizing the activation of gonadotropes. We have developed a novel system to analyze the gonadotrope network which will allow us to study the influence of physiological signals to this interconnected population. To visualize calcium signals specifically in gonadotropes, we created a line of Gonadotrope-GCaMP mice using Gnrhr-driven Cre to express the GCaMP calcium indicator. For present studies, intact pituitary glands were taken from adult Gonadotrope-GCaMP mice (males and proestrous females). Whole pituitaries were mounted in a chamber, and gonadotrope network responses to GnRH (1 or 5nM, at least 4 pituitaries/dose) were recorded using confocal microscopy. Calcium signals from an average of 177 ± 8 gonadotropes per pituitary were analyzed. MATLAB software was used to quantify network connections of gonadotropes for each treatment. Correlation coefficients were determined by their patterns of calcium signaling. Gonadotrope connectivity increases following GnRH stimulation in a dose-dependent manner in female pituitaries (17% following 1nM GnRH, p<0.05; 44% following 5nM GnRH, p<0.05). While male gonadotropes also increase connectivity following stimulation with GnRH, the effect is not enhanced by the higher dose (27% following 1nM GnRH, p<0.05; 20% following 5nM GnRH, NS). Although the strength of gonadotrope correlations is not changed by 1nM GnRH in females, higher dosage of GnRH doubles the strength of correlations over pre-stimulation levels (Pre: 0.08, Post: 0.16, p<0.05). The strength of correlations in males was not significantly altered by GnRH at either dose. In conclusion, the data from whole mouse pituitaries confirm that gonadotropes are capable of high levels of coordinated activity. All pituitaries showed baseline levels of coordination among gonadotropes, confirming the presence of the network. Network activation is significantly increased by GnRH stimulation in both males and females. In females, coordination of gonadotropes is also strengthened by GnRH in a dose-dependent manner. Using a novel gonadotrope-specific calcium indicator mouse model, we have shown GnRH-mediated changes in gonadotrope network connectivity in the intact mouse pituitary. Future studies in our lab will determine how other physiological signals are involved in development and/or maintenance of this coordinated activation. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

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24

Chang, Yao-Chuan, Steven T. Walston, Robert H. Chow y James D. Weiland. "GCaMP expression in retinal ganglion cells characterized using a low-cost fundus imaging system". Journal of Neural Engineering 14, n.º 5 (20 de septiembre de 2017): 056018. http://dx.doi.org/10.1088/1741-2552/aa7ded.

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25

Li, Qiaoran, Libo Yang y Craig Montell. "Drosophila proboscis extension response and GCaMP imaging for assaying food appeal based on grittiness". STAR Protocols 3, n.º 4 (diciembre de 2022): 101806. http://dx.doi.org/10.1016/j.xpro.2022.101806.

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26

Muto, Akira y Koichi Kawakami. "Imaging functional neural circuits in zebrafish with a new GCaMP and the Gal4FF-UAS system". Communicative & Integrative Biology 4, n.º 5 (septiembre de 2011): 566–68. http://dx.doi.org/10.4161/cib.15848.

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27

Cai, Bin, Xia Chen, Fang Liu, Jun Li, Lijuan Gu, Jason R. Liu y Jay Liu. "A Cell-Based Functional Assay Using a Green Fluorescent Protein-Based Calcium Indicator dCys-GCaMP". ASSAY and Drug Development Technologies 12, n.º 6 (agosto de 2014): 342–51. http://dx.doi.org/10.1089/adt.2014.584.

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28

Luo, Jin, Lvli Chen, Feifei Huang, Ping Gao, Heping Zhao, Yingdian Wang y Shengcheng Han. "Intraorganellar calcium imaging in Arabidopsis seedling roots using the GCaMP variants GCaMP6m and R-CEPIA1er". Journal of Plant Physiology 246-247 (marzo de 2020): 153127. http://dx.doi.org/10.1016/j.jplph.2020.153127.

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Collier, Daniel M., Nuria Villalba, Adrian Sackheim, Adrian D. Bonev, Zachary D. Miller, Jesse S. Moore, Bo Shui et al. "Extracellular histones induce calcium signals in the endothelium of resistance-sized mesenteric arteries and cause loss of endothelium-dependent dilation". American Journal of Physiology-Heart and Circulatory Physiology 316, n.º 6 (1 de junio de 2019): H1309—H1322. http://dx.doi.org/10.1152/ajpheart.00655.2018.

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Histone proteins are elevated in the circulation after traumatic injury owing to cellular lysis and release from neutrophils. Elevated circulating histones in trauma contribute to coagulopathy and mortality through a mechanism suspected to involve endothelial cell (EC) dysfunction. However, the functional consequences of histone exposure on intact blood vessels are unknown. Here, we sought to understand the effects of clinically relevant concentrations of histones on the endothelium in intact, resistance-sized, mesenteric arteries (MAs). EC Ca2+ was measured with high spatial and temporal resolution in MAs from mice selectively expressing the EC-specific, genetically encoded ratiometric Ca2+ indicator, Cx40-GCaMP-GR, and vessel diameter was measured by edge detection. Application of purified histone protein directly to the endothelium of en face mouse and human MA preparations produced large Ca2+ signals that spread within and between ECs. Surprisingly, luminal application of histones had no effect on the diameter of pressurized arteries. Instead, after prolonged exposure (30 min), it reduced dilations to endothelium-dependent vasodilators and ultimately caused death of ~25% of ECs, as evidenced by markedly elevated cytosolic Ca2+ levels (793 ± 75 nM) and uptake of propidium iodide. Removal of extracellular Ca2+ but not depletion of intracellular Ca2+ stores prevented histone-induced Ca2+ signals. Histone-induced signals were not suppressed by transient receptor potential vanilloid 4 (TRPV4) channel inhibition (100 nM GSK2193874) or genetic ablation of TRPV4 channels or Toll-like receptor receptors. These data demonstrate that histones are robust activators of noncanonical EC Ca2+ signaling, which cause vascular dysfunction through loss of endothelium-dependent dilation in resistance-sized MAs. NEW & NOTEWORTHY We describe the first use of the endothelial cell (EC)-specific, ratiometric, genetically encoded Ca2+ indicator, Cx40-GCaMP-GR, to study the effect of histone proteins on EC Ca2+ signaling. We found that histones induce an influx of Ca2+ in ECs that does not cause vasodilation but instead causes Ca2+ overload, EC death, and vascular dysfunction in the form of lost endothelium-dependent dilation.

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Fleming, Weston, Sean Jewell, Ben Engelhard, Daniela M. Witten y Ilana B. Witten. "Inferring spikes from calcium imaging in dopamine neurons". PLOS ONE 16, n.º 6 (4 de junio de 2021): e0252345. http://dx.doi.org/10.1371/journal.pone.0252345.

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Calcium imaging has led to discoveries about neural correlates of behavior in subcortical neurons, including dopamine (DA) neurons. However, spike inference methods have not been tested in most populations of subcortical neurons. To address this gap, we simultaneously performed calcium imaging and electrophysiology in DA neurons in brain slices and applied a recently developed spike inference algorithm to the GCaMP fluorescence. This revealed that individual spikes can be inferred accurately in this population. Next, we inferred spikes in vivo from calcium imaging from these neurons during Pavlovian conditioning, as well as during navigation in virtual reality. In both cases, we quantitatively recapitulated previous in vivo electrophysiological observations. Our work provides a validated approach to infer spikes from calcium imaging in DA neurons and implies that aspects of both tonic and phasic spike patterns can be recovered.

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31

Han, Jung Woo, Woon Heo, Donghyuk Lee, Choeun Kang, Hye-Yeon Kim, Ikhyun Jun, Insuk So et al. "Plasma Membrane Localized GCaMP-MS4A12 by Orai1 Co-Expression Shows Thapsigargin- and Ca2+-Dependent Fluorescence Increases". Molecules and Cells 44, n.º 4 (30 de abril de 2021): 223–32. http://dx.doi.org/10.14348/molcells.2021.2031.

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32

Holman, Holly A., Micah D. Frerck y Richard D. Rabbitt. "GCAMP Calcium Imaging Reveals Kinetics and Location of MET Channels in Mammalian Semicircular Canal Hair Cells". Biophysical Journal 114, n.º 3 (febrero de 2018): 287a. http://dx.doi.org/10.1016/j.bpj.2017.11.1647.

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33

Muto, A., M. Ohkura, T. Kotani, S. i. Higashijima, J. Nakai y K. Kawakami. "Genetic visualization with an improved GCaMP calcium indicator reveals spatiotemporal activation of the spinal motor neurons in zebrafish". Proceedings of the National Academy of Sciences 108, n.º 13 (7 de marzo de 2011): 5425–30. http://dx.doi.org/10.1073/pnas.1000887108.

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34

Akerboom, Jasper, Jonathan D. Vélez Rivera, María M. Rodríguez Guilbe, Elisa C. Alfaro Malavé, Hector H. Hernandez, Lin Tian, S. Andrew Hires, Jonathan S. Marvin, Loren L. Looger y Eric R. Schreiter. "Crystal Structures of the GCaMP Calcium Sensor Reveal the Mechanism of Fluorescence Signal Change and Aid Rational Design". Journal of Biological Chemistry 284, n.º 10 (18 de diciembre de 2008): 6455–64. http://dx.doi.org/10.1074/jbc.m807657200.

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35

Sanchez, Colline, Christine Berthier, Bruno Allard y Vincent Jacquemond. "Measurements of Triadic Calcium in Differentiated Muscle Fibers using a Gcamp Probe Targeted to the Junctional Sr Membrane". Biophysical Journal 116, n.º 3 (febrero de 2019): 522a. http://dx.doi.org/10.1016/j.bpj.2018.11.2815.

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36

Rosenthal, Zachary P., Ryan V. Raut, Ping Yan, Deima Koko, Andrew W. Kraft, Leah Czerniewski, Benjamin Acland et al. "Local Perturbations of Cortical Excitability Propagate Differentially Through Large-Scale Functional Networks". Cerebral Cortex 30, n.º 5 (10 de febrero de 2020): 3352–69. http://dx.doi.org/10.1093/cercor/bhz314.

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Abstract Electrophysiological recordings have established that GABAergic interneurons regulate excitability, plasticity, and computational function within local neural circuits. Importantly, GABAergic inhibition is focally disrupted around sites of brain injury. However, it remains unclear whether focal imbalances in inhibition/excitation lead to widespread changes in brain activity. Here, we test the hypothesis that focal perturbations in excitability disrupt large-scale brain network dynamics. We used viral chemogenetics in mice to reversibly manipulate parvalbumin interneuron (PV-IN) activity levels in whisker barrel somatosensory cortex. We then assessed how this imbalance affects cortical network activity in awake mice using wide-field optical neuroimaging of pyramidal neuron GCaMP dynamics as well as local field potential recordings. We report 1) that local changes in excitability can cause remote, network-wide effects, 2) that these effects propagate differentially through intra- and interhemispheric connections, and 3) that chemogenetic constructs can induce plasticity in cortical excitability and functional connectivity. These findings may help to explain how focal activity changes following injury lead to widespread network dysfunction.

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37

Carr, Lynn, Sylvia M. Bardet, Delia Arnaud-Cormos, Philippe Leveque y Rodney P. O'Connor. "Visualisation of an nsPEF induced calcium wave using the genetically encoded calcium indicator GCaMP in U87 human glioblastoma cells". Bioelectrochemistry 119 (febrero de 2018): 68–75. http://dx.doi.org/10.1016/j.bioelechem.2017.09.003.

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38

Pais-Roldán, Patricia, Kengo Takahashi, Filip Sobczak, Yi Chen, Xiaoning Zhao, Hang Zeng, Yuanyuan Jiang y Xin Yu. "Indexing brain state-dependent pupil dynamics with simultaneous fMRI and optical fiber calcium recording". Proceedings of the National Academy of Sciences 117, n.º 12 (5 de marzo de 2020): 6875–82. http://dx.doi.org/10.1073/pnas.1909937117.

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Pupillometry, a noninvasive measure of arousal, complements human functional MRI (fMRI) to detect periods of variable cognitive processing and identify networks that relate to particular attentional states. Even under anesthesia, pupil dynamics correlate with brain-state fluctuations, and extended dilations mark the transition to more arousable states. However, cross-scale neuronal activation patterns are seldom linked to brain state-dependent pupil dynamics. Here, we complemented resting-state fMRI in rats with cortical calcium recording (GCaMP-mediated) and pupillometry to tackle the linkage between brain-state changes and neural dynamics across different scales. This multimodal platform allowed us to identify a global brain network that covaried with pupil size, which served to generate an index indicative of the brain-state fluctuation during anesthesia. Besides, a specific correlation pattern was detected in the brainstem, at a location consistent with noradrenergic cell group 5 (A5), which appeared to be dependent on the coupling between different frequencies of cortical activity, possibly further indicating particular brain-state dynamics. The multimodal fMRI combining concurrent calcium recordings and pupillometry enables tracking brain state-dependent pupil dynamics and identifying unique cross-scale neuronal dynamic patterns under anesthesia.

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39

Asrican, Brent y Juan Song. "Extracting meaningful circuit-based calcium dynamics in astrocytes and neurons from adult mouse brain slices using single-photon GCaMP imaging". STAR Protocols 2, n.º 1 (marzo de 2021): 100306. http://dx.doi.org/10.1016/j.xpro.2021.100306.

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Chen, Yingxiao, Xianqiang Song, Sheng Ye, Lin Miao, Yun Zhu, Rong-Guang Zhang y Guangju Ji. "Structural insight into enhanced calcium indicator GCaMP3 and GCaMPJ to promote further improvement". Protein & Cell 4, n.º 4 (abril de 2013): 299–309. http://dx.doi.org/10.1007/s13238-013-2103-4.

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41

Qiu, Jian, Todd L. Stincic, Martha A. Bosch, Ashley M. Connors, Stefanie Kaech Petrie, Oline K. Rønnekleiv y Martin J. Kelly. "Deletion of Stim1 in Hypothalamic Arcuate Nucleus Kiss1 Neurons Potentiates Synchronous GCaMP Activity and Protects against Diet-Induced Obesity". Journal of Neuroscience 41, n.º 47 (15 de octubre de 2021): 9688–701. http://dx.doi.org/10.1523/jneurosci.0622-21.2021.

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42

Krishnan, Vijai, Lauren C. Wade-Kleyn, Ron R. Israeli y Galit Pelled. "Peripheral Nerve Injury Induces Changes in the Activity of Inhibitory Interneurons as Visualized in Transgenic GAD1-GCaMP6s Rats". Biosensors 12, n.º 6 (1 de junio de 2022): 383. http://dx.doi.org/10.3390/bios12060383.

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Peripheral nerve injury induces cortical remapping that can lead to sensory complications. There is evidence that inhibitory interneurons play a role in this process, but the exact mechanism remains unclear. Glutamate decarboxylase-1 (GAD1) is a protein expressed exclusively in inhibitory interneurons. Transgenic rats encoding GAD1–GCaMP were generated to visualize the activity in GAD1 neurons through genetically encoded calcium indicators (GCaMP6s) in the somatosensory cortex. Forepaw denervation was performed in adult rats, and fluorescent Ca2+ imaging on cortical slices was obtained. Local, intrahemispheric stimulation (cortical layers 2/3 and 5) induced a significantly higher fluorescence change of GAD1-expressing neurons, and a significantly higher number of neurons were responsive to stimulation in the denervated rats compared to control rats. However, remote, interhemispheric stimulation of the corpus callosum induced a significantly lower fluorescence change of GAD1-expressing neurons, and significantly fewer neurons were deemed responsive to stimulation within layer 5 in denervated rats compared to control rats. These results suggest that injury impacts interhemispheric communication, leading to an overall decrease in the activity of inhibitory interneurons in layer 5. Overall, our results provide direct evidence that inhibitory interneuron activity in the deprived S1 is altered after injury, a phenomenon likely to affect sensory processing.

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43

Wang, Maosen, Yi He, Terrence J. Sejnowski y Xin Yu. "Brain-state dependent astrocytic Ca2+ signals are coupled to both positive and negative BOLD-fMRI signals". Proceedings of the National Academy of Sciences 115, n.º 7 (30 de enero de 2018): E1647—E1656. http://dx.doi.org/10.1073/pnas.1711692115.

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Astrocytic Ca2+-mediated gliovascular interactions regulate the neurovascular network in situ and in vivo. However, it is difficult to measure directly both the astrocytic activity and fMRI to relate the various forms of blood-oxygen-level-dependent (BOLD) signaling to brain states under normal and pathological conditions. In this study, fMRI and GCaMP-mediated Ca2+ optical fiber recordings revealed distinct evoked astrocytic Ca2+ signals that were coupled with positive BOLD signals and intrinsic astrocytic Ca2+ signals that were coupled with negative BOLD signals. Both evoked and intrinsic astrocytic calcium signal could occur concurrently or respectively during stimulation. The intrinsic astrocytic calcium signal can be detected globally in multiple cortical sites in contrast to the evoked astrocytic calcium signal only detected at the activated cortical region. Unlike propagating Ca2+ waves in spreading depolarization/depression, the intrinsic Ca2+ spikes occurred simultaneously in both hemispheres and were initiated upon the activation of the central thalamus and midbrain reticular formation. The occurrence of the intrinsic astrocytic calcium signal is strongly coincident with an increased EEG power level of the brain resting-state fluctuation. These results demonstrate highly correlated astrocytic Ca2+ spikes with bidirectional fMRI signals based on the thalamic regulation of cortical states, depicting a brain-state dependency of both astrocytic Ca2+ and BOLD fMRI signals.

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44

Barykina, Natalia V., Vladimir P. Sotskov, Anna M. Gruzdeva, You Kure Wu, Ruben Portugues, Oksana M. Subach, Elizaveta S. Chefanova et al. "FGCaMP7, an Improved Version of Fungi-Based Ratiometric Calcium Indicator for In Vivo Visualization of Neuronal Activity". International Journal of Molecular Sciences 21, n.º 8 (24 de abril de 2020): 3012. http://dx.doi.org/10.3390/ijms21083012.

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Genetically encoded calcium indicators (GECIs) have become a widespread tool for the visualization of neuronal activity. As compared to popular GCaMP GECIs, the FGCaMP indicator benefits from calmodulin and M13-peptide from the fungi Aspergillus niger and Aspergillus fumigatus, which prevent its interaction with the intracellular environment. However, FGCaMP exhibits a two-phase fluorescence behavior with the variation of calcium ion concentration, has moderate sensitivity in neurons (as compared to the GCaMP6s indicator), and has not been fully characterized in vitro and in vivo. To address these limitations, we developed an enhanced version of FGCaMP, called FGCaMP7. FGCaMP7 preserves the ratiometric phenotype of FGCaMP, with a 3.1-fold larger ratiometric dynamic range in vitro. FGCaMP7 demonstrates 2.7- and 8.7-fold greater photostability compared to mEGFP and mTagBFP2 fluorescent proteins in vitro, respectively. The ratiometric response of FGCaMP7 is 1.6- and 1.4-fold higher, compared to the intensiometric response of GCaMP6s, in non-stimulated and stimulated neuronal cultures, respectively. We reveal the inertness of FGCaMP7 to the intracellular environment of HeLa cells using its truncated version with a deleted M13-like peptide; in contrast to the similarly truncated variant of GCaMP6s. We characterize the crystal structure of the parental FGCaMP indicator. Finally, we test the in vivo performance of FGCaMP7 in mouse brain using a two-photon microscope and an NVista miniscope; and in zebrafish using two-color ratiometric confocal imaging.

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45

Bouffard, Jeff, Alyssa D. Cecchetelli, Coleman Clifford, Kriti Sethi, Ronen Zaidel-Bar y Erin J. Cram. "The RhoGAP SPV-1 regulates calcium signaling to control the contractility of theCaenorhabditis elegansspermatheca during embryo transits". Molecular Biology of the Cell 30, n.º 7 (21 de marzo de 2019): 907–22. http://dx.doi.org/10.1091/mbc.e18-10-0633.

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Contractility of the nonmuscle and smooth muscle cells that comprise biological tubing is regulated by the Rho-ROCK (Rho-associated protein kinase) and calcium signaling pathways. Although many molecular details about these signaling pathways are known, less is known about how they are coordinated spatiotemporally in biological tubes. The spermatheca of the Caenorhabditis elegans reproductive system enables study of the signaling pathways regulating actomyosin contractility in live adult animals. The RhoGAP (GTPase-activating protein toward Rho family small GTPases) SPV-1 was previously identified as a negative regulator of RHO-1/Rho and spermathecal contractility. Here, we uncover a role for SPV-1 as a key regulator of calcium signaling. spv-1 mutants expressing the calcium indicator GCaMP in the spermatheca exhibit premature calcium release, elevated calcium levels, and disrupted spatial regulation of calcium signaling during spermathecal contraction. Although RHO-1 is required for spermathecal contractility, RHO-1 does not play a significant role in regulating calcium. In contrast, activation of CDC-42 recapitulates many aspects of spv-1 mutant calcium signaling. Depletion of cdc-42 by RNA interference does not suppress the premature or elevated calcium signal seen in spv-1 mutants, suggesting other targets remain to be identified. Our results suggest that SPV-1 works through both the Rho-ROCK and calcium signaling pathways to coordinate cellular contractility.

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46

Negussie, Mikias, Saritha Krishna y Shawn Hervey-Jumper. "CNTM-02. Regulation of glioma-network integration by tumor mediated secretion of TSP-1". Neuro-Oncology 23, Supplement_6 (2 de noviembre de 2021): vi224. http://dx.doi.org/10.1093/neuonc/noab196.900.

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Abstract Glioma exists in the complex neural circuitry of the brain, making the interface between neoplastic and healthy neurons and glia potentially damaging to long range neural networks and stimulatory to tumor growth. Thrombospondin-1 (TSP-1), an astrocyte derived neurogenic factor expressed by glia of the normal brain, has been found to be upregulated in intratumoral regions with high network functional connectivity (HFC). This modified cell signaling represents cancer cell hijacking of normal physiology with direct impact on tumor biology. There is emerging evidence that neuronal activity influences glioma proliferation and gliomas promote neuronal hyperexcitability. In humans, we have recently shown that bidirectional cellular interactions between gliomas and neurons alter cognitive circuit dynamics and ultimately patient survival. Previously, a subpopulation of human high-grade glioma cells which are enriched for tumor cells with synaptogenic potential were identified (HFC-IHDwtGBM). We plan to study the mechanisms of TSP1 signaling in three different established glioma models (1) HFC-IDHwtGBM hippocampal neuron co-culture, (2) HFC-IDHwtGBM + induced neuron organoids, (3) patient derived xenografts (PDX) for in vivo GCaMP calcium imaging. This project aims to test the hypothesis that increased TSP-1 secretion from HFC-IHDwtGBM cells plays a central role in the maintenance of an invasive and proliferative tumor phenotype when compared with LFC-IHDwtGBM PDX. We hope our study guides future work focused on preventing the infiltration of tumor cells into healthy brain tissues.

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47

Oehler, Beatrice, Cindy Perier, Amy Fisher, Mikhail Kalinichev y Stephen McMahon. "Effects of recombinant botulinum neurotoxin type A1 on CFA-induced mechanical allodynia and sensory neuron responses to mechanical stimulation monitored with GCaMP fluorescence in mice". Toxicon 190 (enero de 2021): S52—S53. http://dx.doi.org/10.1016/j.toxicon.2020.11.452.

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48

Xing, Xiaomin y Chun-Fang Wu. "Unraveling Synaptic GCaMP Signals: Differential Excitability and Clearance Mechanisms Underlying Distinct Ca2+ Dynamics in Tonic and Phasic Excitatory, and Aminergic Modulatory Motor Terminals in Drosophila". eneuro 5, n.º 1 (enero de 2018): ENEURO.0362–17.2018. http://dx.doi.org/10.1523/eneuro.0362-17.2018.

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49

Xing, Xiaomin y Chun-Fang Wu. "Inter-relationships among physical dimensions, distal–proximal rank orders, and basal GCaMP fluorescence levels in Ca2+ imaging of functionally distinct synaptic boutons at Drosophila neuromuscular junctions". Journal of Neurogenetics 32, n.º 3 (3 de julio de 2018): 195–208. http://dx.doi.org/10.1080/01677063.2018.1504043.

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50

Frost, Nicholas A., Anna Haggart y Vikaas S. Sohal. "Dynamic patterns of correlated activity in the prefrontal cortex encode information about social behavior". PLOS Biology 19, n.º 5 (3 de mayo de 2021): e3001235. http://dx.doi.org/10.1371/journal.pbio.3001235.

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New technologies make it possible to measure activity from many neurons simultaneously. One approach is to analyze simultaneously recorded neurons individually, then group together neurons which increase their activity during similar behaviors into an “ensemble.” However, this notion of an ensemble ignores the ability of neurons to act collectively and encode and transmit information in ways that are not reflected by their individual activity levels. We used microendoscopic GCaMP imaging to measure prefrontal activity while mice were either alone or engaged in social interaction. We developed an approach that combines a neural network classifier and surrogate (shuffled) datasets to characterize how neurons synergistically transmit information about social behavior. Notably, unlike optimal linear classifiers, a neural network classifier with a single linear hidden layer can discriminate network states which differ solely in patterns of coactivity, and not in the activity levels of individual neurons. Using this approach, we found that surrogate datasets which preserve behaviorally specific patterns of coactivity (correlations) outperform those which preserve behaviorally driven changes in activity levels but not correlated activity. Thus, social behavior elicits increases in correlated activity that are not explained simply by the activity levels of the underlying neurons, and prefrontal neurons act collectively to transmit information about socialization via these correlations. Notably, this ability of correlated activity to enhance the information transmitted by neuronal ensembles is diminished in mice lacking the autism-associated gene Shank3. These results show that synergy is an important concept for the coding of social behavior which can be disrupted in disease states, reveal a specific mechanism underlying this synergy (social behavior increases correlated activity within specific ensembles), and outline methods for studying how neurons within an ensemble can work together to encode information.

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