Construction of a two-photon microscope for video-rate Ca2+ imaging

Nguyen, QT; Callamaras, Nick; Hsieh, Candace Y.; Parker, Ian

doi:10.1054/ceca.2001.0246

Cited by 116 publications

(98 citation statements)

References 20 publications

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“…Ca 2ϩ imaging and flash photolysis. Imaging was performed using a custom-made video-rate two-photon microscope, as described previously (Nguyen et al, 2001;Stutzmann et al, 2003;Stutzmann and Parker, 2005). In brief, excitation was provided by trains (80 MHz) of ϳ100 fs pulses at 780 nm from a Ti:sapphire laser (Tsunami; Spectra-Physics, Mountain View, CA).…”

Section: Methodsmentioning

confidence: 99%

Enhanced Ryanodine Receptor Recruitment Contributes to Ca²⁺Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice

Stutzmann¹,

Smith²,

Caccamo³

et al. 2006

J. Neurosci.

313

324

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Neuronal Ca2ϩ signaling through inositol triphosphate receptors (IP 3 R) and ryanodine receptors (RyRs) must be tightly regulated to maintain cell viability, both acutely and over a lifetime. Exaggerated intracellular Ca 2ϩ levels have been associated with expression of Alzheimer's disease (AD) mutations in young mice, but little is known of Ca 2ϩ dysregulations during normal and pathological aging processes. Here, we used electrophysiological recordings with two-photon imaging to study Ca 2ϩ signaling in nontransgenic (NonTg) and several AD mouse models (PS1 KI , 3xTg-AD, and APP Swe Tau P301L ) at young (6 week), adult (6 months), and old (18 months) ages. At all ages, the PS1 KI and 3xTg-AD mice displayed exaggerated endoplasmic reticulum (ER) Ca 2ϩ signals relative to NonTg mice. The PS1 mutation was the predominant "calciopathic" factor, because responses in 3xTg-AD mice were similar to PS1 KI mice , and APP Swe Tau P301L mice were not different from controls. In addition, we uncovered powerful signaling interactions and differences between IP 3 R-and RyR-mediated Ca 2ϩ components in NonTg and AD mice. In NonTg mice, RyR contributed modestly to IP 3 -evoked Ca 2ϩ , whereas the exaggerated signals in 3xTg-AD and PS1 KI mice resulted primarily from enhanced RyR-Ca 2ϩ release and were associated with increased RyR expression across all ages. Moreover, IP 3 -evoked membrane hyperpolarizations in AD mice were even greater than expected from exaggerated Ca 2ϩ signals, suggesting increased coupling efficiency between cytosolic [Ca 2ϩ ] and K ϩ channel regulation. We conclude that lifelong ER Ca 2ϩ disruptions in AD are related to a modulation of RyR signaling associated with PS1 mutations and represent a discrete "calciumopathy," not merely an acceleration of normal aging.

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Section: Methodsmentioning

confidence: 99%

Enhanced Ryanodine Receptor Recruitment Contributes to Ca²⁺Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice

Stutzmann¹,

Smith²,

Caccamo³

et al. 2006

J. Neurosci.

313

324

View full text Add to dashboard Cite

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“…1, 2). For that purpose, the combination of high-speed raster scanning (Leybaert and Sanderson, 2001;Nguyen et al, 2001;Rochefort et al, 2009;Jia et al, 2010;Cheng et al, 2011), principal component analysis (Mitra and Pesaran, 1999;Mukamel et al, 2009), and nonlinear deconvolution (Vogelstein et al, 2010) was particularly powerful. Because each frame is scanned in 30 ms and because physiological motion from, for example, heartbeat or breathing is small and comparatively slow (Յ10 Hz), motion in x-y is approximately rigid and can be corrected with 2D registration.…”

Section: Random Stimuli For Population Imagingmentioning

confidence: 99%

Local Diversity and Fine-Scale Organization of Receptive Fields in Mouse Visual Cortex

Bonin

Histed²,

Yurgenson³

et al. 2011

J. Neurosci.

242

217

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Many thousands of cortical neurons are activated by any single sensory stimulus, but the organization of these populations is poorly understood. For example, are neurons in mouse visual cortex-whose preferred orientations are arranged randomly-organized with respect to other response properties? Using high-speed in vivo two-photon calcium imaging, we characterized the receptive fields of up to 100 excitatory and inhibitory neurons in a 200 m imaged plane. Inhibitory neurons had nonlinearly summating, complex-like receptive fields and were weakly tuned for orientation. Excitatory neurons had linear, simple receptive fields that can be studied with noise stimuli and system identification methods. We developed a wavelet stimulus that evoked rich population responses and yielded the detailed spatial receptive fields of most excitatory neurons in a plane. Receptive fields and visual responses were locally highly diverse, with nearby neurons having largely dissimilar receptive fields and response time courses. Receptive-field diversity was consistent with a nearly random sampling of orientation, spatial phase, and retinotopic position. Retinotopic positions varied locally on average by approximately half the receptive-field size. Nonetheless, the retinotopic progression across the cortex could be demonstrated at the scale of 100 m, with a magnification of ϳ10 m/°. Receptive-field and response similarity were in register, decreasing by 50% over a distance of 200 m. Together, the results indicate considerable randomness in local populations of mouse visual cortical neurons, with retinotopy as the principal source of organization at the scale of hundreds of micrometers.

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“…Imaging was performed with a custom-built, two-photon microscope system as described, except that three photomultiplier channels were used for simultaneous imaging of green-, red-, and blue-labeled lymphocytes (18,19). The system was based on an upright Olympus BX50 microscope fitted with a ϫ20 water-immersion objective (numerical aperture 0.95), a Spectra-Physics Tsunami femtosecond laser tuned to 780 nm, an x-y mirror scan head, and a motorized focus controller (Prior Scientific).…”

Section: Two-photon Imaging and Data Analysismentioning

confidence: 99%

Class IA Phosphoinositide 3-Kinase Modulates Basal Lymphocyte Motility in the Lymph Node

Matheu¹,

Deane²,

Parker

et al. 2007

The Journal of Immunology

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Recruitment of PI3K to the cell membrane is an indispensable step in normal lymphocyte proliferation and activation. In this study we identify PI3K as an important signaling molecule for maintaining basal T and B lymphocyte motility and homing in the intact lymph node. Pharmacological inhibition of PI3K catalytic isoforms exerted broad effects on basal lymphocyte motility, including changes in homing kinetics, localization of B cells within the lymph node, and reduced cell velocities. Lymphocytes deficient in either or both of the class IA PI3K regulatory subunits p85α and p85β also exhibited reduced velocities, with the magnitude of reduction depending upon both cell type and isoform specificity. B cells deficient in p85α exhibited gross morphological abnormalities that were not evident in cells treated with a PI3K inhibitor. Our results show, for the first time, that class IA PI3Ks play an important role in regulating basal lymphocyte motility and that p85α regulatory subunit expression is required to maintain B cell morphology in a manner independent of PI3K catalytic function. Moreover, we demonstrate distinct roles for catalytic domain function and class IA PI3K regulatory domain activity in lymphocyte motility, homing, and homeostatic localization of mature resting B cells.

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Construction of a two-photon microscope for video-rate Ca2+ imaging

Cited by 116 publications

References 20 publications

Enhanced Ryanodine Receptor Recruitment Contributes to Ca²⁺Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice

Enhanced Ryanodine Receptor Recruitment Contributes to Ca²⁺Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice

Local Diversity and Fine-Scale Organization of Receptive Fields in Mouse Visual Cortex

Class IA Phosphoinositide 3-Kinase Modulates Basal Lymphocyte Motility in the Lymph Node

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Construction of a two-photon microscope for video-rate Ca2+ imaging

Cited by 116 publications

References 20 publications

Enhanced Ryanodine Receptor Recruitment Contributes to Ca2+Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice

Enhanced Ryanodine Receptor Recruitment Contributes to Ca2+Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice

Local Diversity and Fine-Scale Organization of Receptive Fields in Mouse Visual Cortex

Class IA Phosphoinositide 3-Kinase Modulates Basal Lymphocyte Motility in the Lymph Node

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Product

Resources

About

Enhanced Ryanodine Receptor Recruitment Contributes to Ca²⁺Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice

Enhanced Ryanodine Receptor Recruitment Contributes to Ca²⁺Disruptions in Young, Adult, and Aged Alzheimer's Disease Mice