Thy1-GCaMP6 Transgenic Mice for Neuronal Population Imaging In Vivo

Dana, Hod; Chen, Tsai Wen; Hu, Amy; Shields, Brenda C.; Guo, Caiying; Looger, Loren L.; Kim, Douglas S.; Svoboda, Karel

doi:10.1371/journal.pone.0108697

Cited by 540 publications

(648 citation statements)

References 36 publications

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“…Because photodamage can be mediated by fluorophore excitation, we conducted these experiments with both wild-type mice and GP4.12 (GCaMP6S) transgenic mice (Dana et al 2014). As with weakly focused light, increased immunoreactivity was evident at powers of 300 mW and above in both mouse strains following 20-min continuous illumination (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Brain heating induced by near-infrared lasers during multiphoton microscopy

Podgorski

Ranganathan

2016

Journal of Neurophysiology

244

234

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Podgorski K, Ranganathan G. Brain heating induced by nearinfrared lasers during multiphoton microscopy. J Neurophysiol 116: 1012-1023, 2016. First published June 8, 2016 doi:10.1152/jn.00275.2016.-Two-photon imaging and optogenetic stimulation rely on high illumination powers, particularly for stateof-the-art applications that target deeper structures, achieve faster measurements, or probe larger brain areas. However, little information is available on heating and resulting damage induced by high-power illumination in the brain. In the current study we used thermocouple probes and quantum dot nanothermometers to measure temperature changes induced by two-photon microscopy in the neocortex of awake and anaesthetized mice. We characterized heating as a function of wavelength, exposure time, and distance from the center of illumination. Although total power is highest near the surface of the brain, heating was most severe hundreds of micrometers below the focal plane, due to heat dissipation through the cranial window. Continuous illumination of a 1-mm 2 area produced a peak temperature increase of ϳ1.8°C/100 mW. Continuous illumination with powers above 250 mW induced lasting damage, detected with immunohistochemistry against Iba1, glial fibrillary acidic protein, heat shock proteins, and activated caspase-3. Higher powers were usable in experiments with limited duty ratios, suggesting an approach to mitigate damage in high-power microscopy experiments.

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Section: Resultsmentioning

confidence: 99%

“…One experiment (see Fig. 6) involved GP4.12 mice (Dana et al 2014). Both male and female mice, between 8 and 22 wk of age, were included in studies.…”

Section: Methodsmentioning

confidence: 99%

Brain heating induced by near-infrared lasers during multiphoton microscopy

Podgorski

Ranganathan

2016

Journal of Neurophysiology

244

234

View full text Add to dashboard Cite

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“…Importantly, the imaging paradigm presented here should be broadly applicable to other GCaMP6 mouse lines that are developed, such as those that do not require a separate driver line (Dana et al 2014) or those where expression is dependent on Cre recombinase (Madisen et al 2015). In addition, because this line is driven by the tetO system, it provides an orthogonal targeting scheme to Cre control that may be useful for other imaging approaches.…”

Section: Discussionmentioning

confidence: 99%

Large-scale imaging of cortical dynamics during sensory perception and behavior

Wekselblatt

Flister

Piscopo

et al. 2016

Journal of Neurophysiology

265

316

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Sensory-driven behaviors engage a cascade of cortical regions to process sensory input and generate motor output. To investigate the temporal dynamics of neural activity at this global scale, we have improved and integrated tools to perform functional imaging across large areas of cortex using a transgenic mouse expressing the genetically encoded calcium sensor GCaMP6s, together with a head-fixed visual discrimination behavior. This technique allows imaging of activity across the dorsal surface of cortex, with spatial resolution adequate to detect differential activity in local regions at least as small as 100 μm. Imaging during an orientation discrimination task reveals a progression of activity in different cortical regions associated with different phases of the task. After cortex-wide patterns of activity are determined, we demonstrate the ability to select a region that displayed conspicuous responses for two-photon microscopy and find that activity in populations of individual neurons in that region correlates with locomotion in trained mice. We expect that this paradigm will be a useful probe of information flow and network processing in brain-wide circuits involved in many sensory and cognitive processes.

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“…These indicators reliably detect rapid calcium transients in various neural cell types in vivo. [24][25][26] Using cultured PTE cells, we performed in vitro confocal microscopy studies; using two-photon microscopy, we examined the in vivo calcium modulations in renal PT cells. The in vitro experiments properly identified the cell types examined and allowed a detailed, properly calibrated examination of the effects of ligands and potential inhibitors on the PT epithelial cellular calcium levels.…”

Section: Discussionmentioning

confidence: 99%

Visualization of Calcium Dynamics in Kidney Proximal Tubules

Szebényi

Füredi

Kolacsek

et al. 2015

Journal of the American Society of Nephrology

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Intrarenal changes in cytoplasmic calcium levels have a key role in determining pathologic and pharmacologic responses in major kidney diseases. However, cell-specific delivery of calcium-sensitive probes in vivo remains problematic. We generated a transgenic rat stably expressing the green fluorescent protein-calmodulinbased genetically encoded calcium indicator (GCaMP2) predominantly in the kidney proximal tubules. The transposon-based method used allowed the generation of homozygous transgenic rats containing one copy of the transgene per allele with a defined insertion pattern, without genetic or phenotypic alterations. We applied in vitro confocal and in vivo two-photon microscopy to examine basal calcium levels and ligand-and drug-induced alterations in these levels in proximal tubular epithelial cells. Notably, renal ischemia induced a transient increase in cellular calcium, and reperfusion resulted in a secondary calcium load, which was significantly decreased by systemic administration of specific blockers of the angiotensin receptor and the Na-Ca exchanger. The parallel examination of in vivo cellular calcium dynamics and renal circulation by fluorescent probes opens new possibilities for physiologic and pharmacologic investigations.

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Thy1-GCaMP6 Transgenic Mice for Neuronal Population Imaging In Vivo

Cited by 540 publications

References 36 publications

Brain heating induced by near-infrared lasers during multiphoton microscopy

Brain heating induced by near-infrared lasers during multiphoton microscopy

Large-scale imaging of cortical dynamics during sensory perception and behavior

Visualization of Calcium Dynamics in Kidney Proximal Tubules

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