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Literatura académica sobre el tema "GCaMP"

Autor: Grafiati

Publicado: 3 de julio de 2021

Última modificación: 10 de marzo de 2023

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Artículos de revistas sobre el tema "GCaMP"

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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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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Introduction: 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. Methods: 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. Results: 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. Conclusion: 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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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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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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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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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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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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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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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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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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Tesis sobre el tema "GCaMP"

Pereira, Lucas Borges. "Caracterização da apirase do parasita P. falciparum e análise do papel do Ca2+ no egresso de T. gondii". Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/42/42135/tde-17082016-151526/.

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Plasmodium falciparum e Toxoplasma gondii são protozoários parasitas pertencentes ao filo Apicomplexa. Apirases são enzimas metabolizadoras de nucleotídeos extracelulares. Nesta tese mostramos pela primeira vez a presença de um membro desta família de enzimas em P. falciparum, o qual foi capaz de degradar ATP extracelular. Análises por RT-qPCR revelaram a expressão da apirase durante todo o ciclo intraeritrocítico. A adição de inibidores desta classe de enzimas foi capaz de prejudicar o desenvolvimento dos parasitas e a invasão de novas hemácias pelos merozoitos, sugerindo assim um papel da apirase nestes processos. A via de sinalização por Ca2+ é universal e vital para todas as células. Para melhor entender a fisiologia celular de P. falciparum construímos uma nova linhagem de parasitas transgênicos, PfGCaMP3, que nos tornam capazes de monitorar a dinâmica de Ca2+ sem o uso de protocolos invasivos de marcação. De modo semelhante utilizamos uma nova linhagem de T. gondii expressando de forma estável o indicador de Ca2+ GCaMP3 para estudar o papel deste íon na saída da célula. T. gondii possui o Ca2+ necessário para promover este processo, entretanto Ca2+ extracelular age como um fator intensificador neste passo essencial do ciclo lítico.
Plasmodium falciparum and Toxoplasma gondii are protozoan parasites that belong to phylum Apicomplexa. Apirases are metabolizing enzymes of extracellular nucleotides. In this work we show for the first time the presence of an apyrase in P. falciparum, which was able to degrade extracellular ATP. RTqPCR analysis revealed the expression of apyrase throughout the intraerythrocytic cycle. Addition of apyrase inhibitors was able to impair the development of the parasites and the invasion of new erythrocytes by merozoites, thus suggesting a role of apyrase in these processes. Calcium signaling is universal and vital to all cells. To better understand the cellular physiology of P. falciparum we construct a new strain of transgenic parasites, PfGCaMP3, which enable us to monitor the Ca2+ dynamics without using invasive protocols. Similarly we use a new strain of T. gondii that stably express the Ca2+ indicator GCaMP3 to study the role Ca2+ in parasite egress. T. gondii has the Ca2+ required to promote this process, however extracellular Ca2+ acts as an enhancer factor in this crucial step of the lytic cycle.

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Iguchi, Moritake. "Direct monitoring of mitochondrial calcium levels in cultured cardiac myocytes using a novel fluorescent indicator protein, GCaMP2-mt". Kyoto University, 2011. http://hdl.handle.net/2433/142548.

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Schmidt, Elke. "Investigation of spatiotemporal calcium transients in astrocytic soma and processes upon purinergic receptor activation using genetically encoded calcium sensors". Thesis, Sorbonne Paris Cité, 2015. http://www.theses.fr/2015PA05T011.

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Les astrocytes protoplasmiques de la matière grise corticale sont des cellules gliales dont les prolongements très fins et ramifiés sont en contact avec les éléments neuronaux pré- et post-synaptiques d’une part, et les vaisseaux sanguins d’autre part. Ils expriment plusieurs récepteurs des neurotransmetteurs, entre autres des récepteurs purinergiques dont l'activation facilite l’activité calcique astrocytaire et la libération de gliotransmitters (par exemple, le glutamate, le GABA, l'ATP, et la D sérine) qui régulent l’activité des neurones et des cellules gliales situées au voisinage. L’objectif de ma thèse était d’étudier in situ l’activité calcique des astrocytes et de leurs prolongements en réponse à l’application des agonistes purinergiques. Lors de ma thèse, j’ai tout d'abord testé la possibilité d’induire l’expression spécifique de gènes d’intérêt par les astrocytes corticaux de souris adultes par la technique de recombinaison Cre-LoxP. J’ai comparé les performances d’un virus adeno-associé de type 5 (AAV5) flexé (AAV5.FLEX.EGFP) et d’une souris qui exprime un indicateur calcique (GCaMP3) sous contrôle de la recombinase (souris Rosa-CAG-LSL-GCaMP3). L’injection d’AAV5.FLEX.EGFP dans le cortex d’une souris hGFAPcre n’a pas permis l’expression spécifique d’EGFP. La combinaison des souris exprimant le cre recombinase sous contrôle d’un promoteur sélectif des astrocytes (GLAST-CreERT2 et Cx30-CreERT2) avec le AAV5.FLEX.EGFP ou avec une lignée des souris Rosa-CAG-LSL-GCaMP3 permet l’expression spécifique des gènes d’intérêt (EGFP et GCaMP3) par les astrocytes corticaux. J’ai ensuite analysé l’activité calcique des astrocytes qui expriment GCaMP3. J’ai utilisé la microscopie biphotonique et enregistré l’activité calcique spontanée et évoquée par application d’agonistes purinergiques sur des tranches de cortex somatosensoriel primaire de souris adultes GLAST-CreERT2. L’activité calcique spontanée est complexe, généralement locale et désynchronisée, répartie dans les prolongements et la région somatique. Les régions actives ont été identifiées à partir d’une carte de corrélation temporale calculée en MATLAB, et leurs caractéristiques (amplitude, durée, position, fréquence) mesurées grâce à des routines établies sous IGOR. La fréquence et l’amplitude de l’activité calcique paraissent augmenter lors de l’enregistrement, ce qui suggère une sensibilité significative et une photoactivation des astrocytes, en imagerie biphotonique. La durée des impulsions laser modulerait ce phénomène. En présence d'adénosine (1-100 µM) et d’ATP (100 µM), et de façon marginale en présence d’un agoniste P2X7 non sélectif (BzATP 50-100 µM), une activité calcique synchronisée accrue est visible dans le soma et les prolongements astrocytaires en présence de tétrodotoxine qui bloque les potentiels d'action et minimise l’activité synaptique. Le mécanisme de ces réponses synchronisées reste à étudier. Aucun effet significatif n’a été observé en présence d’un agoniste spécifique P2Y1 (MRS2365 50 uM). Mon travail a permis le développement : i) de modèles murins pour l’adressage sélectif de protéines d’intérêt au niveau des astrocytes protoplasmiques ; ii) d’outils d’analyse des signaux calciques astrocytaires au niveau sub-cellulaire. Il a mis en évidence des limites possibles des protocoles standards d'enregistrement de l’activité calcique des astrocytes en imagerie biphotonique. Il confirme l’importance de l’ATP et de l’adénosine pour la signalisation astrocytaire
Grey matter protoplasmic astrocytes are compact glial cells with highly branched processes, enwrapping synapses, and one or two endfeet contacting the blood vessels. Several neurotransmitter receptors are expressed by astrocytes, among them purinergic receptors. Upon activation of these receptors, intracellular calcium (Ca2+) transients can be induced, that, in turn, trigger gliotransmitter release (e.g. glutamate, GABA, ATP, D-serine) and participate in astrocyte-to-astrocyte signaling as well as in the communication between astrocytes and neurons or other glia. During my PhD work, I first implemented and validated several approaches for targeting transgene expression specifically to cortical astrocytes and employed them to study purinergic signaling in astrocytes. To achieve astrocyte-specific transgene expression, I used either floxed adeno-associated viral (AAV) vectors or a Cre-dependent mouse line and several mouse lines expressing the Cre recombinase under astrocyte-specific promoters. Intracerebral injections of a Cre-dependent AAV serotype 5 containing the ubiquitous CAG promoter and an enhanced green fluorescent protein (AAV5.CAG.flex.EGFP) in adult mice expressing Cre recombinase under the human glial fibrillary protein (hGFAP) promoter resulted in a non-astrocyte specific expression in the cortex. Combining inducible mouse lines expressing Cre recombinase under the glutamate aspartate transporter (GLAST) promoter with the same AAV vector resulted in a virtually astrocyte-specific expression of the reporter gene. As an alternative approach for astrocyte-specific transgene expression, we used a Cre-dependent mouse line expressing the genetically encoded Ca2+ indicator GCaMP3. Crossing this mouse line with the above described GLAST-CreERT2 mouse line or a Connexin30 (Cx30)-CreERT2 line led to selective GCaMP3 expression in cortical astrocytes. Second, I investigated both spontaneous and agonist-evoked Ca2+ transients in astrocytic processes, the investigation of which has presented a major challenge in earlier studies, due to the unspecific and weak labeling by membrane-permeable chemical Ca2+ indicators. Using the strategy developed in the first part of my work allowing an astrocyte-specific expression of the genetically encoded Ca2+ indicator GCaMP3. Using two-photon excitation fluorescence (2PEF) imaging in acute slices of the primary somatosensory cortex, I recorded Ca2+ transients in the astrocytic soma and processes. By aid of a custom-made MATLAB routine based on a temporal Pearson correlation coefficient, active regions could be identified in an unbiased manner. Evoked Ca2+ transients were quantified using custom IGOR routines. Spontaneous desynchronized Ca2+ transients occurred in the processes and rarely in the soma. Ca2+ signals appeared localized in distinct microdomains. Their frequency appeared to increase during long recordings of several hundred images, suggesting that fine astrocytes are vulnerable to photodamage under imaging conditions routine in 2PEF microscopy. The possibility to minimize photodamage, by varying the length of the femtosecond laser pulses is under investigation. Bath application of adenosine (1-100 µM) and adenosine-triphosphate (ATP, 100 µM), as well as the application of the non-selective P2X7 receptor agonist (2'(3')-O-(4-Benzoylbenzoyl)adenosine-5'-triphosphate, BzATP, 50-100 µM), in the presence of tetrodotoxin to block neuronal action potentials, evoked synchronized Ca2+ rises in the soma and the processes of astrocytes. The effect of adenosine was dose-dependent. No significant effect of the specific P2Y1 agonist (MRS2365, 50 µM) was seen. Altogether, my work sets up a powerful and versatile toolbox for studying astrocytic Ca2+ signaling at the sub-cellular level. It also pinpoints possible limits of standard two-photon recording protocols to investigate the local Ca2+ signals in fine astrocytic processes

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Schröder, Thomas [Verfasser], Hauke [Akademischer Betreuer] Lilie, Daniel [Akademischer Betreuer] Huster y Karl-Wilhelm [Akademischer Betreuer] Koch. "Konformationsänderung des Guanylatzyklase-aktivierenden Proteins 2 (GCAP-2) zur Aktivierung der Sehstäbchenaußensegment-Guanylatzyklase (ROS-GC1) / Thomas Schröder. Betreuer: Hauke Lilie ; Daniel Huster ; Karl-Wilhelm Koch". Halle, Saale : Universitäts- und Landesbibliothek Sachsen-Anhalt, 2011. http://d-nb.info/102513513X/34.

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Matthus, Elsa. "Phosphate starvation alters calcium signalling in roots of Arabidopsis thaliana". Thesis, University of Cambridge, 2019. https://www.repository.cam.ac.uk/handle/1810/290260.

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Low bioavailability of phosphate (P) due to low concentration and high immobility in soils is a key limiting factor in crop production. Application of excess amounts of P fertilizer is costly and by no means sustainable, as world-wide P resources are finite and running out. To facilitate the breeding of crops adapted to low-input soils, it is essential to understand the consequences of P deficiency. The second messenger calcium (Ca2+) is known to signal in plant development and stress perception, and most recently its direct role in signalling nutrient availability and deficiency has been partially elucidated. The use of Ca2+ as a signal has to be tightly controlled, as Ca2+ easily complexes with P groups and therefore is highly toxic to cellular P metabolism. It is unknown whether Ca2+ signals P availability or whether signalling is altered under P starvation conditions. The aim of this PhD project was to characterise the use of Ca2+ ions, particularly cytosolic free Ca2+ ([Ca2+]cyt), in stress signalling by P-starved roots of the model plant Arabidopsis thaliana. The hypothesis was that under P starvation and a resulting decreased cellular P pool, the use of [Ca2+]cyt may have to be restricted to avoid cytotoxic complexation of Ca2+ with limited P groups. Employing a range of genetically encoded Ca2+ reporters in Arabidopsis, P starvation but not nitrogen starvation was found to strongly dampen the root [Ca2+]cyt increases evoked by mechanical, salt, osmotic, and oxidative stress as well as by extracellular nucleotides. The strongly altered root [Ca2+]cyt response to extracellular nucleotides was shown to manifest itself during seedling development under chronic P deprivation, but could be reversed by P resupply. Fluorescent imaging elucidated that P-starved roots showed a normal [Ca2+]cyt response to extracellular nucleotides at the apex, but a strongly dampened [Ca2+]cyt response in distal parts of the root tip, correlating with high reactive oxygen species (ROS) levels induced by P starvation. Excluding iron, as well as P, rescued the altered [Ca2+]cyt response, and restored ROS levels to those seen under nutrient-replete conditions. P availability was not signalled through [Ca2+]cyt. In another part of this PhD project, a library of 77 putative Ca2+ channel mutants was compiled and screened for aberrant root hair growth under P starvation conditions. No mutant line showed aberrant root hair growth. These results indicate that P starvation strongly affects stress-induced [Ca2+]cyt modulations. The data generated in this thesis further understanding of how plants can integrate nutritional and environmental cues, adding another layer of complexity to the use of Ca2+ as a signal transducer.

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Gualtieri, Charles J. "Sensory Representation of Social Stimuli in Aromatase Expressing Neurons in the Medial Amygdala". 2021. https://scholarworks.umass.edu/masters_theses_2/1050.

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The ability of animals to sense, interpret, and respond appropriately to social stimuli in their environment is essential for identifying and distinguishing between members of their own species. In mammals, social interactions both within and across species play a key role in determining if an animal will live to pass on its genes to the next generation or else be removed from the gene pool. The result of this selection pressure can be observed in specialized neural circuits that respond to social stimuli and orchestrate appropriate behavioral responses. This highly conserved network of brain structures is often referred to as the Social Behavior Network (SBN). The medial amygdala (MeA) is a central node in the SBN and has been shown to be involved in transforming information from olfactory sensory systems into social and defensive behavioral responses. Previous research has shown that individual neurons in the MeA of anesthetized mice respond selectively to different chemosensory social cues, a characteristic not observed in its upstream relay, the accessory olfactory bulb (AOB). However, the cause of this stimulus selectivity in the MeA is not yet understood. Here, I hypothesize that a subpopulation of neurons in the MeA that express the enzyme aromatase are involved in the sensory representation of social stimuli in awake, behaving animals. To test this hypothesis, I designed and built a novel behavioral apparatus that allows for discrete presentations of social stimuli in a highly controllable and reproducible environment. I then injected the adeno-associated virus (AAV) AAV-Syn-Flex-GCAMP6s into the MeA of Aromatase:Cre transgenic mice and implanted a fiber optic cannula slightly above the injection site. The combination of this transgenic mouse line and conditional AAV caused GCaMP6s expression to be exclusive to aromatase-expressing neurons. By coupling my novel behavioral apparatus to a fiber photometry system, I successfully recorded the moment-to-moment activity of aromatase neurons in the MeA of awake, behaving animals as they investigated various social stimuli. Aromatase neurons in the MeA of adult male mice respond strongly to conspecific social stimuli, including live adult mice, mouse pups, and mouse urine samples. Sniffing and investigative behaviors correlated strongly with increased GCaMP6s signal in aromatase neurons, reflecting increases in their neural activity. Interestingly, after repeated investigations of the same stimuli the activity of aromatase neurons gradually diminished. Presenting a novel stimulus following repeated investigations of a familiar stimulus reinstated some, but not all of the initial GCaMP6s signal. This points to the potential role that aromatase neurons may play in the habituation to social stimuli that are consistently present in their environment. Investigations of predator stimuli did not evoke significant responses from aromatase neurons, nor did investigations of non-social stimuli. These results demonstrate that aromatase expressing neurons in the MeA of awake, behaving animals encode the sensory representation of conspecific social stimuli, and their responses are highly selective to the type of stimulus presented.

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Turrini, Lapo. "Development of optical methods for real-time whole-brain functional imaging of zebrafish neuronal activity". Doctoral thesis, 2019. http://hdl.handle.net/2158/1152459.

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In this PhD thesis, we performed functional imaging of zebrafish larvae with a multi-modal and multi-scale approach. In particular, we focused our research on measuring neuronal activity in both physiological and pathological conditions. We adopted a pharmacological model of epilepsy, by administering pentylentetrazole at different concentrations and inducing seizures of different entity in zebrafish larvae expressing in all CNS neurons the Ca2+ reporter GCaMP6s. Owing to the relation between neuronal activity (i.e. action potentials) and intracellular Ca2+ concentration, we were able to measure neuronal activity by recording the changes in fluorescence of GCaMP indicator. Using a custom-made widefield fluorescence microscope, we measured activity during the onset and propagation of seizures, investigating the dynamics between different brain regions along with tail locomotor activity. We implemented commonly used zebrafish high-throughput drug screening assays, measuring only behavioural parameters (i.e. velocity of swimming, total travelled length), with a direct measure of the overall brain activity, thus laying the foundation for novel drug screening methods capable of improved efficacy. In order to improve the spatio-temporal resolution of brain activity recordings, being able to perform optical sectioning of the transparent zebrafish brain, we performed Bessel beam illumination light-sheet fluorescence microscopy measurements. Indeed, with respect to conventional Gaussian illumination, Bessel beams, owing to their nondiffractive and self-healing properties, allow for a substantial reduction of haemodynamic artefacts jeopardizing functional recordings in conventional measurements. We applied a custom analysis pipeline to produce 3D maps of neuronal activity, with single cell resolution. Finally, in order to perform real-time whole-brain measurements with singleneuron resolution, we devised a novel two-photon light-sheet microscope able to image the entire larval brain at 1 Hz. Applying a pixel-wise custom analysis we were able to identify functional circuitries involved both in physiological and pathological neuronal communication.

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Gilyan, Andrew. "Optimizing Genetically Encoded Calcium Indicators to Measure Presynaptic Calcium Transients". 2012. http://hdl.handle.net/10222/50610.

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Neurotransmitter release is modulated by multiple regulatory mechanisms that control several stages of synaptic vesicle (SV) exocytosis. At the final stage, SV fusion with the presynaptic membrane requires calcium influx through voltage-gated calcium channels, and regulatory mechanisms that alter the surface expression or conductance of calcium channels have large effects on neurotransmitter release. To determine how these mechanisms contribute to synapse-specific modulations of neurotransmitter release and synaptic strength, we require a means to monitor presynaptic calcium transients at individual synapses. Genetically encoded calcium indicators (GECIs), engineered proteins that change their fluorescence emission properties upon calcium binding, generally lack the sensitivity to measure such transients in response to isolated stimuli. Therefore, we modified the GECI, GCaMP3, by altering its sensitivity for calcium. Our results suggest the modified GCaMP-based presynaptically targeted GECIs are excellent tools to quantify presynaptic calcium transients at individual synapses in response to isolated action potentials.

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"Visual Analytics Tool for the Global Change Assessment Model". Master's thesis, 2015. http://hdl.handle.net/2286/R.I.35998.

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abstract: The Global Change Assessment Model (GCAM) is an integrated assessment tool for exploring consequences and responses to global change. However, the current iteration of GCAM relies on NetCDF file outputs which need to be exported for visualization and analysis purposes. Such a requirement limits the uptake of this modeling platform for analysts that may wish to explore future scenarios. This work has focused on a web-based geovisual analytics interface for GCAM. Challenges of this work include enabling both domain expert and model experts to be able to functionally explore the model. Furthermore, scenario analysis has been widely applied in climate science to understand the impact of climate change on the future human environment. The inter-comparison of scenario analysis remains a big challenge in both the climate science and visualization communities. In a close collaboration with the Global Change Assessment Model team, I developed the first visual analytics interface for GCAM with a series of interactive functions to help users understand the simulated impact of climate change on sectors of the global economy, and at the same time allow them to explore inter comparison of scenario analysis with GCAM models. This tool implements a hierarchical clustering approach to allow inter-comparison and similarity analysis among multiple scenarios over space, time, and multiple attributes through a set of coordinated multiple views. After working with this tool, the scientists from the GCAM team agree that the geovisual analytics tool can facilitate scenario exploration and enable scientific insight gaining process into scenario comparison. To demonstrate my work, I present two case studies, one of them explores the potential impact that the China south-north water transportation project in the Yangtze River basin will have on projected water demands. The other case study using GCAM models demonstrates how the impact of spatial variations and scales on similarity analysis of climate scenarios varies at world, continental, and country scales.
Dissertation/Thesis
Masters Thesis Computer Science 2015

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Mati, Jacob Mwathi. "Global civil society advocacy alliances and networks in the changing terrain of global governance and development : a critical inquiry into the politics and dynamics in crafting and operations of the Global Action against Poverty (GCAP)". Thesis, 2009. http://hdl.handle.net/10539/6102.

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The last few decades have witnessed the emergence of global civil society advocacy networks as major players in global governance. This research aimed at using a case study of GCAP in critically analysing the politics and dynamics of crafting a global civil society advocacy alliance. Specifically, the study aimed to: a) identify, analyse, and document GCAP’s experiences, strategies and challenges in trans-national networking and advocacy; b) generate knowledge on recent developments in global civil society networking and advocacy. The study analyses the study phenomenon using two central features of GCAP: networking and advocacy. Chapter one attempts to give a background of the study and also discusses the methods used. Chapter two lays the theoretical framework and operationalises the concepts explored in the study. The report argues that alliances are very different from ‘normal’ forms of organisations because they are made up of diverse forms of organisations, coming together voluntarily to achieve a specific purpose. They are therefore, by their very nature, complex, unstable, and difficult to co-ordinate. Chapters Three and Four look at such intricacies and complexities of crafting and operations of global advocacy networks. I conclude this research arguing that despite challenges in alliances building and operations, global civil society organisations will still need to network if they are to remain relevant and effective in current global governance context. It is only in their unity that they will be able to confront their common challenges.

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Libros sobre el tema "GCaMP"

J, Foose Thomas y IUCN/SSC Captive Breeding Specialist Group., eds. Rhino global captive action plan (GCAP): 1 September 1992. [S.l.]: IUCN/SSC Captive Breeding Specialist Group, 1992.

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Cinzia, Virno, ed. GCAMC: Roma, Galleria comunale d'arte moderna e contemporanea : catalogo generale delle collezioni : autori dell'Ottocento. Roma: F.lli Palombi, 2004.

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Cinzia, Virno y Bonasegale Pittei Giovanna, eds. GCAMC: Roma, Galleria comunale d'arte moderna e contemporanea : catalogo generale delle collezioni : autori dell'Ottocento. Roma: F.lli Palombi, 2004.

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Capítulos de libros sobre el tema "GCaMP"

Anil Sethi, Anjula Garg, Jan Sacharko, Regina List, Jesse O. Bollinger, Lisiunia Romanienko, Allyson Reaves, David B. Howard et al. "GCAP". En International Encyclopedia of Civil Society, 751. New York, NY: Springer US, 2010. http://dx.doi.org/10.1007/978-0-387-93996-4_9115.

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Baehr, Wolfgang, Iswari Subbaraya, Wojciech A. Gorczyca y Krzysztof Palczewski. "Guanylate Cyclase-Activating Protein (GCAP)". En Degenerative Diseases of the Retina, 339–47. Boston, MA: Springer US, 1995. http://dx.doi.org/10.1007/978-1-4615-1897-6_38.

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Koch, Karl-Wilhelm. "GCAP (Guanylate Cyclase–Activating Protein)". En Encyclopedia of Signaling Molecules, 2041–45. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-67199-4_12.

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Koch, Karl-Wilhelm. "GCAP (Guanylate Cyclase–Activating Protein)". En Encyclopedia of Signaling Molecules, 1–5. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4614-6438-9_12-1.

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Meigs, Thomas E., Alex Lyakhovich, Hoon Shim, Ching-Kang Chen, Denis J. Dupré, Terence E. Hébert, Joe B. Blumer et al. "GCAP (Guanylate Cyclase–Activating Protein)". En Encyclopedia of Signaling Molecules, 769–73. New York, NY: Springer New York, 2012. http://dx.doi.org/10.1007/978-1-4419-0461-4_12.

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Anil Sethi, Anjula Garg, Jan Sacharko, Regina List, Jesse O. Bollinger, Lisiunia Romanienko, Allyson Reaves, David B. Howard et al. "Global Call to Action Against Poverty (GCAP)". En International Encyclopedia of Civil Society, 769–70. New York, NY: Springer US, 2010. http://dx.doi.org/10.1007/978-0-387-93996-4_759.

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Diaz, Michel, Roberto Canonico, Luis Costa, Serge Fdida, David Hutchison, Laurent Mathy, Andreas Meissner, Stephane Owezarski, Rolland Vida y Lars Wolf. "GCAP: A New Multimedia Multicast Architecture for QoS". En Protocols for Multimedia Systems, 103–15. Berlin, Heidelberg: Springer Berlin Heidelberg, 2001. http://dx.doi.org/10.1007/3-540-45481-0_9.

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Mendez, Ana y Jeannie Chen. "Mouse Models to Study GCAP Functions In Intact Photoreceptors". En Advances in Experimental Medicine and Biology, 361–88. Boston, MA: Springer US, 2002. http://dx.doi.org/10.1007/978-1-4615-0121-3_22.

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Zammali, Saloua, Khedija Arour y Amel Bouzeghoub. "GCAPM: A Generic Context-Aware Model in Peer-to-Peer Environment". En Lecture Notes in Computer Science, 364–71. Berlin, Heidelberg: Springer Berlin Heidelberg, 2013. http://dx.doi.org/10.1007/978-3-642-40173-2_29.

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Ames, B. y Mitsuhiko Ikura. "Structure and Membrane-Targeting Mechanism of Retinal Ca2+-Binding Proteins, Recoverin and GCAP-2". En Advances in Experimental Medicine and Biology, 333–48. Boston, MA: Springer US, 2002. http://dx.doi.org/10.1007/978-1-4615-0121-3_20.

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Actas de conferencias sobre el tema "GCaMP"

Hubert, Antoine, Fabrice Harms, Sophia Imperato, Vincent Loriette, Cynthia Veilly, Xavier Levecq, Georges Farkouh, François Rouyer y Alexandra Fragola. "Adaptive Optics Light-Sheet Microscopy for Functional Neuroimaging". En European Conference on Biomedical Optics. Washington, D.C.: Optica Publishing Group, 2021. http://dx.doi.org/10.1364/ecbo.2021.em2b.1.

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We present a new implementation of adaptive optics for light-sheet microscopy, with a direct extended-scene wavefront sensing measurement for fast aberration correction. We report AO-enhanced images of GCaMP in freshly dissected drosophila brains.

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Ciuparu, Andrei y Raul C. Muresan. "Jittered sampling - a potential solution for detecting high frequencies in GCaMP recordings". En 2021 IEEE 17th International Conference on Intelligent Computer Communication and Processing (ICCP). IEEE, 2021. http://dx.doi.org/10.1109/iccp53602.2021.9733598.

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Perez-Zoghbi, J. F., G. T. Yocum y C. W. Emala. "Intracellular Calcium Dynamics in Murine Airway Smooth Muscle Studied with a GCaMP Probe". En American Thoracic Society 2021 International Conference, May 14-19, 2021 - San Diego, CA. American Thoracic Society, 2021. http://dx.doi.org/10.1164/ajrccm-conference.2021.203.1_meetingabstracts.a4331.

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Zhu, Xiaobo. "Computer Simulation on Global Atmospheric Transport of Mercury by GEOS-Chem/GCAP". En 2018 3rd International Conference on Modelling, Simulation and Applied Mathematics (MSAM 2018). Paris, France: Atlantis Press, 2018. http://dx.doi.org/10.2991/msam-18.2018.21.

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Brondi, Marco, Manuel Molano-Mazón, Stefano Panzeri y Tommaso Fellin. "High Accuracy Two-Photon Population Imaging of GCaMP6 Signals with Fast Smart Line Scan". En Optics and the Brain. Washington, D.C.: OSA, 2018. http://dx.doi.org/10.1364/brain.2018.bw2c.5.

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Najafizadeh, Laleh, David Margolis, Li Zhu y Christian Lee. "Probing the dynamics of spontaneous cortical activities via widefield Ca+2 imaging in GCaMP6 transgenic mice". En Wavelets and Sparsity XVII, editado por Yue M. Lu, Manos Papadakis y Dimitri Van De Ville. SPIE, 2017. http://dx.doi.org/10.1117/12.2274119.

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Wang, Tianyu, Dimitre G. Ouzounov, Mengran Wang, Danielle Feng, Jean C. Cruz-Hernandez, Jacob Reimer, Andreas Tolias, Nozomi Nishimura y Chris Xu. "In vivo three-photon activity imaging of GCaMP6-labeled neurons in deep cortex and the hippocampus of the mouse brain". En SPIE BiOS, editado por Ammasi Periasamy, Peter T. C. So, Karsten König y Xiaoliang S. Xie. SPIE, 2017. http://dx.doi.org/10.1117/12.2251220.

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Futia, Gregory L., Arjun Fontaine, Samuel Littich, Connor McCullough, Diego Restrepo, Richard Weir, John Caldwell y Emily A. Gibson. "In vivo holographic photo-stimulation and two photon GCaMP6 imaging of vagus nerve axons using a GRIN lens integrated nerve cuff". En Optogenetics and Optical Manipulation 2019, editado por Samarendra K. Mohanty y E. Duco Jansen. SPIE, 2019. http://dx.doi.org/10.1117/12.2521830.

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Lazarou, Stavros, Christos Christodoulou y Vasiliki Vita. "Global Change Assessment Model (GCAM) considerations of the primary sources energy mix for an energetic scenario that could meet Paris agreement". En 2019 54th International Universities Power Engineering Conference (UPEC). IEEE, 2019. http://dx.doi.org/10.1109/upec.2019.8893507.

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Informes sobre el tema "GCaMP"

Smith, S. J., A. H. Mizrahi, J. F. Karas y M. Nathan. US Renewable Futures in the GCAM. Office of Scientific and Technical Information (OSTI), octubre de 2011. http://dx.doi.org/10.2172/1219303.

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Smith, Steven J., Andrew H. Mizrahi, Joseph F. Karas y Mayda Nathan. US Renewable Futures in the GCAM. Office of Scientific and Technical Information (OSTI), octubre de 2011. http://dx.doi.org/10.2172/1027702.

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Dooley, James J. y Yuyu Zhou. Explicitly Accounting for Protected Lands within the GCAM 3.0. Office of Scientific and Technical Information (OSTI), mayo de 2012. http://dx.doi.org/10.2172/1068653.

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Smith, Steven J., April C. Volke y Sabrina Delgado Arias. Enhancement of Solar Energy Representation in the GCAM Model. Office of Scientific and Technical Information (OSTI), febrero de 2010. http://dx.doi.org/10.2172/1033089.

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Kyle, G. Page, Patrick Luckow, Katherine V. Calvin, William R. Emanuel, Mayda Nathan y Yuyu Zhou. GCAM 3.0 Agriculture and Land Use: Data Sources and Methods. Office of Scientific and Technical Information (OSTI), diciembre de 2011. http://dx.doi.org/10.2172/1036082.

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Hannam, Phil, G. Page Kyle y Steven J. Smith. Global Deployment of Geothermal Energy Using a New Characterization in GCAM 1.0. Office of Scientific and Technical Information (OSTI), septiembre de 2009. http://dx.doi.org/10.2172/991595.

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Edmonds, J. A., M. A. Wise y C. N. MacCracken. ADVANCED ENERGY TECHNOLOGIES AND CLIMATE CHANGE: AN ANALYSIS USING THE GLOBAL CHANGE ASSESSMENT MODEL (GCAM). Office of Scientific and Technical Information (OSTI), mayo de 1994. http://dx.doi.org/10.2172/1127203.

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Binsted, Matthew, Harry Suchyta, Ying Zhang, Laura Vimmerstedt, Matt Mowers, Catherine Ledna, Matteo Muratori y Chioke Harris. Renewable Energy and Efficiency Technologies in Scenarios of U.S. Decarbonization in Two Types of Models: Comparison of GCAM Modeling and Sector-Specific Modeling. Office of Scientific and Technical Information (OSTI), diciembre de 2022. http://dx.doi.org/10.2172/1903177.

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