Fluorescence Changes of Genetic Calcium Indicators and OGB-1 Correlated with Neural Activity and Calcium<i>In Vivo</i>and<i>In Vitro</i>

Hendel, Thomas; Mank, Marco; Schnell, Bettina; Griesbeck, Oliver; Borst, Alexander; Reiff, Dierk F.

doi:10.1523/jneurosci.1038-08.2008

Cited by 167 publications

(134 citation statements)

References 69 publications

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“…Our finding is congruent with previous observations that the same cameleon may show different K d values in different subcellular compartments (14,32). One potential limitation of D3cpv is the slower kinetics compared with the Ca 2ϩ fluorescent dyes (19), so it may miss the detection of very rapid transient events. However, D3cpv has been used successfully to detect single Ca 2ϩ spikes in neuronal cells (54) sparks and regenerative global Ca 2ϩ release through crossactivation of ryanodine receptors (61).…”

Section: Et-1 and Ang II Preferentially Triggered Subsarcolemmal Ca 2supporting

confidence: 92%

“…6E) 2ϩ of D3cpv and 3NLS-D3cpv determined in situ are similar, but they are slightly lower than the K d of 0.6 M determined in vitro using recombinant D3cpv (38). This is in agreement with earlier findings on several types of cameleons showing a decrease in the K d value when calibrated in vivo compared with in vitro (19). In contrast, the PM-targeted Lyn-D3cpv has a slightly higher in situ K d compared with those of D3cpv and 3NLS-D3cpv.…”

Section: Et-1 and Ang II Preferentially Triggered Subsarcolemmal Ca 2supporting

confidence: 91%

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Detection of differentially regulated subsarcolemmal calcium signals activated by vasoactive agonists in rat pulmonary artery smooth muscle cells

Subedi

Paudel

Sham

2014

American Journal of Physiology-Cell Physiology

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Section: Et-1 and Ang II Preferentially Triggered Subsarcolemmal Ca 2supporting

confidence: 92%

Section: Et-1 and Ang II Preferentially Triggered Subsarcolemmal Ca 2supporting

confidence: 91%

Detection of differentially regulated subsarcolemmal calcium signals activated by vasoactive agonists in rat pulmonary artery smooth muscle cells

Subedi

Paudel

Sham

2014

American Journal of Physiology-Cell Physiology

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“…G-CaMP family of calcium sensors have been used in a variety of animal systems and their signals have been shown to correlate well with calcium transients [25][26][27][28] . The use of genetically-encoded sensor provides a sensitive readout of the neuronal responses and allows specific expression of the sensor in targeted tissue and cell type.…”

Section: Discussionmentioning

confidence: 99%

Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor

Haga-Yamanaka

et al. 2011

JoVE

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The vomeronasal organ (VNO) detects chemosensory signals that carry information about the social, sexual and reproductive status of the individuals within the same species 1,2 . These intraspecies signals, the pheromones, as well as signals from some predators 3 , activate the vomeronasal sensory neurons (VSNs) with high levels of specificity and sensitivity 4 . At least three distinct families of G-protein coupled receptors, V1R, , are expressed in VNO neurons to mediate the detection of the chemosensory cues. To understand how pheromone information is encoded by the VNO, it is critical to analyze the response profiles of individual VSNs to various stimuli and identify the specific receptors that mediate these responses.The neuroepithelia of VNO are enclosed in a pair of vomer bones. The semi-blind tubular structure of VNO has one open end (the vomeronasal duct) connecting to the nasal cavity. VSNs extend their dendrites to the lumen part of the VNO, where the pheromone cues are in contact with the receptors expressed at the dendritic knobs. The cell bodies of the VSNs form pseudo-stratified layers with V1R and V2R expressed in the apical and basal layers respectively 6-8 . Several techniques have been utilized to monitor responses of VSNs to sensory stimuli 4,12,[15][16][17][18][19] . Among these techniques, acute slice preparation offers several advantages. First, compared to dissociated VSNs 3,17 , slice preparations maintain the neurons in their native morphology and the dendrites of the cells stay relatively intact. Second, the cell bodies of the VSNs are easily accessible in coronal slice of the VNO to allow electrophysiology studies and imaging experiments as compared to whole epithelium and whole-mount preparations 12,20 . Third, this method can be combined with molecular cloning techniques to allow receptor identification.Sensory stimulation elicits strong Ca 2+ influx in VSNs that is indicative of receptor activation 4,21 . We thus develop transgenic mice that express G-CaMP2 in the olfactory sensory neurons, including the VSNs 15,22 . The sensitivity and the genetic nature of the probe greatly facilitate Ca 2+ imaging experiments. This method has eliminated the dye loading process used in previous studies 4,21 . We also employ a ligand delivery system that enables application of various stimuli to the VNO slices. The combination of the two techniques allows us to monitor multiple neurons simultaneously in response to large numbers of stimuli. Finally, we have established a semi-automated analysis pipeline to assist image processing.

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“…In addition, by using tissue specific promoters and selective cellular targeting modifications, these proteins became most useful for calcium signaling analysis within selected tissues, specific cell types or intracellular compartments (see Refs. [13][14][15][16][17]). In most cases, under proper expression conditions, GECIs have minimum effects on cellular activity, can be used in vivo as well, and therefore are also suitable for long-term studies (see Ref.…”

Section: Methods For Studying Calcium Signals In Human Ps Cellsmentioning

confidence: 99%

Calcium signaling in human pluripotent stem cells

2016

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a b s t r a c tHuman pluripotent stem cells provide new tools for developmental and pharmacological studies as well as for regenerative medicine applications. Calcium homeostasis and ligand-dependent calcium signaling are key components of major cellular responses, including cell proliferation, differentiation or apoptosis. Interestingly, these phenomena have not been characterized in detail as yet in pluripotent human cell sates. Here we review the methods applicable for studying both short-and long-term calcium responses, focusing on the expression of fluorescent calcium indicator proteins and imaging methods as applied in pluripotent human stem cells. We discuss the potential regulatory pathways involving calcium responses in hPS cells and compare these to the implicated pathways in mouse PS cells. A recent development in the stem cell field is the recognition of so called "naïve" states, resembling the earliest potential forms of stem cells during development, as well as the "fuzzy" stem cells, which may be alternative forms of pluripotent cell types, therefore we also discuss the potential role of calcium homeostasis in these PS cell types.

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Fluorescence Changes of Genetic Calcium Indicators and OGB-1 Correlated with Neural Activity and CalciumIn VivoandIn Vitro

Cited by 167 publications

References 69 publications

Detection of differentially regulated subsarcolemmal calcium signals activated by vasoactive agonists in rat pulmonary artery smooth muscle cells

Detection of differentially regulated subsarcolemmal calcium signals activated by vasoactive agonists in rat pulmonary artery smooth muscle cells

Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor

Calcium signaling in human pluripotent stem cells

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