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https://www.readbyqxmd.com/read/27922063/design-and-mechanistic-insight-into-ultrafast-calcium-indicators-for-monitoring-intracellular-calcium-dynamics
#1
Nordine Helassa, Borbala Podor, Alan Fine, Katalin Török
Calmodulin-based genetically encoded fluorescent calcium indicators (GCaMP-s) are powerful tools of imaging calcium dynamics from cells to freely moving animals. High affinity indicators with slow kinetics however distort the temporal profile of calcium transients. Here we report the development of reduced affinity ultrafast variants of GCaMP6s and GCaMP6f. We hypothesized that GCaMP-s have a common kinetic mechanism with a rate-limiting process in the interaction of the RS20 peptide and calcium-calmodulin...
December 6, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27847865/in-vivo-characterization-of-distinct-modality-specific-subsets-of-somatosensory-neurons-using-gcamp
#2
Edward C Emery, Ana P Luiz, Shafaq Sikandar, Rán Magnúsdóttir, Xinzhong Dong, John N Wood
Mechanistic insights into pain pathways are essential for a rational approach to treating this vast and increasing clinical problem. Sensory neurons that respond to tissue damage (nociceptors) may evoke pain sensations and are typically classified on the basis of action potential velocity. Electrophysiological studies have suggested that most of the C-fiber nociceptors are polymodal, responding to a variety of insults. In contrast, gene deletion studies in the sensory neurons of transgenic mice have frequently resulted in modality-specific deficits...
November 2016: Science Advances
https://www.readbyqxmd.com/read/27809317/a-microfluidics-based-method-for-measuring-neuronal-activity-in-drosophila-chemosensory-neurons
#3
Lena van Giesen, G Larisa Neagu-Maier, Jae Young Kwon, Simon G Sprecher
Monitoring neuronal responses to defined sensory stimuli is a powerful and widely used approach for understanding sensory coding in the nervous system. However, providing precise, stereotypic and reproducible cues while concomitantly recording neuronal activity remains technically challenging. Here we describe the fabrication and use of a microfluidics system that allows precise temporally restricted stimulation of Drosophila chemosensory neurons with an array of different chemical cues. The system can easily be combined with genetically encoded calcium sensors, and it can measure neuronal activity at single-cell resolution in larval sense organs and in the proboscis or leg of the adult fly...
December 2016: Nature Protocols
https://www.readbyqxmd.com/read/27725685/generation-of-transgenic-marmosets-expressing-genetically-encoded-calcium-indicators
#4
Jung Eun Park, Xian Feng Zhang, Sang-Ho Choi, Junko Okahara, Erika Sasaki, Afonso C Silva
Chronic monitoring of neuronal activity in the living brain with optical imaging techniques became feasible owing to the continued development of genetically encoded calcium indicators (GECIs). Here we report for the first time the successful generation of transgenic marmosets (Callithrix jacchus), an important nonhuman primate model in neurophysiological research, which were engineered to express the green fluorescent protein (GFP)-based family of GECIs, GCaMP, under control of either the CMV or the hSyn promoter...
October 11, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27464819/calcium-imaging-of-neuronal-activity-in-free-swimming-larval-zebrafish
#5
Akira Muto, Koichi Kawakami
Visualization of neuronal activity during animal behavior is a critical step in understanding how the brain generates behavior. In the model vertebrate zebrafish, imaging of the brain has been done mostly by using immobilized fish. Here, we describe a novel method to image neuronal activity of the larval zebrafish brain during prey capture behavior. We expressed a genetically encoded fluorescent calcium indicator, GCaMP, in the optic tectum of the midbrain using the Gal4-UAS system. Tectal activity was then imaged in unrestrained larvae during prey perception...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27441501/calcium-imaging-with-genetically-encoded-indicators-in-behaving-primates
#6
Eyal Seidemann, Yuzhi Chen, Yoon Bai, Spencer C Chen, Preeti Mehta, Bridget L Kajs, Wilson S Geisler, Boris V Zemelman
Understanding the neural basis of behaviour requires studying brain activity in behaving subjects using complementary techniques that measure neural responses at multiple spatial scales, and developing computational tools for understanding the mapping between these measurements. Here we report the first results of widefield imaging of genetically encoded calcium indicator (GCaMP6f) signals from V1 of behaving macaques. This technique provides a robust readout of visual population responses at the columnar scale over multiple mm(2) and over several months...
2016: ELife
https://www.readbyqxmd.com/read/27383588/transient-oxygen-glucose-deprivation-causes-a-delayed-loss-of-mitochondria-and-increases-spontaneous-calcium-signaling-in-astrocytic-processes
#7
John C O'Donnell, Joshua G Jackson, Michael B Robinson
UNLABELLED: Recently, mitochondria have been localized to astrocytic processes where they shape Ca(2+) signaling; this relationship has not been examined in models of ischemia/reperfusion. We biolistically transfected astrocytes in rat hippocampal slice cultures to facilitate fluorescent confocal microscopy, and subjected these slices to transient oxygen/glucose deprivation (OGD) that causes delayed excitotoxic death of CA1 pyramidal neurons. This insult caused a delayed loss of mitochondria from astrocytic processes and increased colocalization of mitochondria with the autophagosome marker LC3B...
July 6, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/27380932/in-vivo-transplantation-of-fetal-human-gut-derived-enteric-neural-crest-cells
#8
J E Cooper, D Natarajan, C J McCann, S Choudhury, H Godwin, A J Burns, N Thapar
The prospect of using neural cell replacement for the treatment of severe enteric neuropathies has seen significant progress in the last decade. The ability to harvest and transplant enteric neural crest cells (ENCCs) that functionally integrate within recipient intestine has recently been confirmed by in vivo murine studies. Although similar cells can be harvested from human fetal and postnatal gut, no studies have as yet verified their functional viability upon in vivo transplantation. We sought to determine whether ENCCs harvested from human fetal bowel are capable of engraftment and functional integration within recipient intestine following in vivo transplantation into postnatal murine colon...
July 6, 2016: Neurogastroenterology and Motility: the Official Journal of the European Gastrointestinal Motility Society
https://www.readbyqxmd.com/read/27335411/cell-type-specific-modulation-of-sensory-responses-in-olfactory-bulb-circuits-by-serotonergic-projections-from-the-raphe-nuclei
#9
Daniela Brunert, Yusuke Tsuno, Markus Rothermel, Michael T Shipley, Matt Wachowiak
UNLABELLED: Serotonergic neurons in the brainstem raphe nuclei densely innervate the olfactory bulb (OB), where they can modulate the initial representation and processing of olfactory information. Serotonergic modulation of sensory responses among defined OB cell types is poorly characterized in vivo Here, we used cell-type-specific expression of optical reporters to visualize how raphe stimulation alters sensory responses in two classes of GABAergic neurons of the mouse OB glomerular layer, periglomerular (PG) and short axon (SA) cells, as well as mitral/tufted (MT) cells carrying OB output to piriform cortex...
June 22, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/27335402/rapid-postnatal-expansion-of-neural-networks-occurs-in-an-environment-of-altered-neurovascular-and-neurometabolic-coupling
#10
Mariel G Kozberg, Ying Ma, Mohammed A Shaik, Sharon H Kim, Elizabeth M C Hillman
UNLABELLED: In the adult brain, increases in neural activity lead to increases in local blood flow. However, many prior measurements of functional hemodynamics in the neonatal brain, including functional magnetic resonance imaging (fMRI) in human infants, have noted altered and even inverted hemodynamic responses to stimuli. Here, we demonstrate that localized neural activity in early postnatal mice does not evoke blood flow increases as in the adult brain, and elucidate the neural and metabolic correlates of these altered functional hemodynamics as a function of developmental age...
June 22, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/26933036/using-a-genetically-encoded-sensor-to-identify-inhibitors-of-toxoplasma-gondii-ca2-signaling
#11
Saima M Sidik, Miryam A Hortua Triana, Aditya S Paul, Majida El Bakkouri, Caroline G Hackett, Fanny Tran, Nicholas J Westwood, Raymond Hui, William J Zuercher, Manoj T Duraisingh, Silvia N J Moreno, Sebastian Lourido
The life cycles of apicomplexan parasites progress in accordance with fluxes in cytosolic Ca(2+) Such fluxes are necessary for events like motility and egress from host cells. We used genetically encoded Ca(2+) indicators (GCaMPs) to develop a cell-based phenotypic screen for compounds that modulate Ca(2+) signaling in the model apicomplexan Toxoplasma gondii In doing so, we took advantage of the phosphodiesterase inhibitor zaprinast, which we show acts in part through cGMP-dependent protein kinase (protein kinase G; PKG) to raise levels of cytosolic Ca(2+) We define the pool of Ca(2+) regulated by PKG to be a neutral store distinct from the endoplasmic reticulum...
April 29, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/26863447/calcium-imaging-of-neuronal-activity-in-drosophila-can-identify-anticonvulsive-compounds
#12
Anne K Streit, Yuen Ngan Fan, Laura Masullo, Richard A Baines
Although there are now a number of antiepileptic drugs (AEDs) available, approximately one-third of epilepsy patients respond poorly to drug intervention. The reasons for this are complex, but are probably reflective of the increasing number of identified mutations that predispose individuals to this disease. Thus, there is a clear requirement for the development of novel treatments to address this unmet clinical need. The existence of gene mutations that mimic a seizure-like behaviour in the fruit fly, Drosophila melanogaster, offers the possibility to exploit the powerful genetics of this insect to identify novel cellular targets to facilitate design of more effective AEDs...
2016: PloS One
https://www.readbyqxmd.com/read/26851502/neuron-class-specific-requirements-for-fragile-x-mental-retardation-protein-in-critical-period-development-of-calcium-signaling-in-learning-and-memory-circuitry
#13
Caleb A Doll, Kendal Broadie
Neural circuit optimization occurs through sensory activity-dependent mechanisms that refine synaptic connectivity and information processing during early-use developmental critical periods. Fragile X Mental Retardation Protein (FMRP), the gene product lost in Fragile X syndrome (FXS), acts as an activity sensor during critical period development, both as an RNA-binding translation regulator and channel-binding excitability regulator. Here, we employ a Drosophila FXS disease model to assay calcium signaling dynamics with a targeted transgenic GCaMP reporter during critical period development of the mushroom body (MB) learning/memory circuit...
May 2016: Neurobiology of Disease
https://www.readbyqxmd.com/read/26439313/ziram-a-pesticide-associated-with-increased-risk-for-parkinson-s-disease-differentially-affects-the-presynaptic-function-of-aminergic-and-glutamatergic-nerve-terminals-at-the-drosophila-neuromuscular-junction
#14
Ciara A Martin, Katherine M Myers, Audrey Chen, Nathan T Martin, Angel Barajas, Felix E Schweizer, David E Krantz
Multiple populations of aminergic neurons are affected in Parkinson's disease (PD), with serotonergic and noradrenergic loci responsible for some non-motor symptoms. Environmental toxins, such as the dithiocarbamate fungicide ziram, significantly increase the risk of developing PD and the attendant spectrum of both motor and non-motor symptoms. The mechanisms by which ziram and other environmental toxins increase the risk of PD, and the potential effects of these toxins on aminergic neurons, remain unclear...
January 2016: Experimental Neurology
https://www.readbyqxmd.com/read/26387442/itches-stimulating-compounds-from-colocasia-esculenta-taro-bioactive-guided-screening-and-lc-ms-ms-identification
#15
Jin-Gao Yu, Pei Liu, Jin-Ao Duan, Zong-Xiang Tang, Yan Yang
Colocasia esculenta (L.) Schoot (taro) is one of the most common crops in the world. Its rhizome was a tonic medicine and accustomed to treat some gastrointestinal disorders in traditional Chinese medicine. Today, the taro was further developed as anticancer prescription in herbal therapy. However, the mucilage of the fresh taro has irritation, and causes itchy feeling. The components in the mucilage were not evident up to now. Two active compounds, uracil and glycol-protein taro lectin (Accession number: A5HMM7), were purified and identified from the fresh taro...
October 15, 2015: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/26382605/green-to-red-photoconversion-of-gcamp
#16
Minrong Ai, Holly Mills, Makoto Kanai, Jason Lai, Jingjing Deng, Eric Schreiter, Loren Looger, Thomas Neubert, Greg Suh
Genetically encoded calcium indicators (GECIs) permit imaging intracellular calcium transients. Among GECIs, the GFP-based GCaMPs are the most widely used because of their high sensitivity and rapid response to changes in intracellular calcium concentrations. Here we report that the fluorescence of GCaMPs--including GCaMP3, GCaMP5 and GCaMP6--can be converted from green to red following exposure to blue-green light (450-500 nm). This photoconversion occurs in both insect and mammalian cells and is enhanced in a low oxygen environment...
2015: PloS One
https://www.readbyqxmd.com/read/26379509/prenatal-and-acute-cocaine-exposure-affects-neural-responses-and-habituation-to-visual-stimuli
#17
Elizabeth Riley, Konstantin Kopotiyenko, Irina Zhdanova
Psychostimulants have many effects on visual function, from adverse following acute and prenatal exposure to therapeutic on attention deficit. To determine the impact of prenatal and acute cocaine exposure on visual processing, we studied neuronal responses to visual stimuli in two brain regions of a transgenic larval zebrafish expressing the calcium indicator GCaMP-HS. We found that both red light (LF) and dark (DF) flashes elicited similar responses in the optic tectum neuropil (TOn), while the dorsal telencephalon (dTe) responded only to LF...
2015: Frontiers in Neural Circuits
https://www.readbyqxmd.com/read/26022075/putative-microcircuit-level-substrates-for-attention-are-disrupted-in-mouse-models-of-autism
#18
Francisco J Luongo, Meryl E Horn, Vikaas S Sohal
BACKGROUND: Deep layer excitatory circuits in the prefrontal cortex represent the strongest locus for genetic convergence in autism, but specific abnormalities within these circuits that mediate key features of autism, such as cognitive or attentional deficits, remain unknown. Attention normally increases the sensitivity of neural populations to incoming signals by decorrelating ongoing cortical circuit activity. Here, we investigated whether mechanisms underlying this phenomenon might be disrupted within deep layer prefrontal circuits in mouse models of autism...
April 15, 2016: Biological Psychiatry
https://www.readbyqxmd.com/read/25989524/in-vivo-two-photon-imaging-of-axonal-dieback-blood-flow-and-calcium-influx-with-methylprednisolone-therapy-after-spinal-cord-injury
#19
Peifu Tang, Yiling Zhang, Chao Chen, Xinran Ji, Furong Ju, Xingyu Liu, Wen-Biao Gan, Zhigang He, Shengxiang Zhang, Wei Li, Lihai Zhang
Severe spinal cord injury (SCI) can cause neurological dysfunction and paralysis. However, the early dynamic changes of neurons and their surrounding environment after SCI are poorly understood. Although methylprednisolone (MP) is currently the standard therapeutic agent for treating SCI, its efficacy remains controversial. The purpose of this project was to investigate the early dynamic changes and MP's efficacy on axonal damage, blood flow, and calcium influx into axons in a mouse SCI model. YFP H-line and Thy1-GCaMP transgenic mice were used in this study...
May 19, 2015: Scientific Reports
https://www.readbyqxmd.com/read/25970202/monitoring-brain-activity-with-protein-voltage-and-calcium-sensors
#20
Douglas A Storace, Oliver R Braubach, Lei Jin, Lawrence B Cohen, Uhna Sung
Understanding the roles of different cell types in the behaviors generated by neural circuits requires protein indicators that report neural activity with high spatio-temporal resolution. Genetically encoded fluorescent protein (FP) voltage sensors, which optically report the electrical activity in distinct cell populations, are, in principle, ideal candidates. Here we demonstrate that the FP voltage sensor ArcLight reports odor-evoked electrical activity in the in vivo mammalian olfactory bulb in single trials using both wide-field and 2-photon imaging...
May 13, 2015: Scientific Reports
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