Aβ Plaques Lead to Aberrant Regulation of Calcium Homeostasis In Vivo Resulting in Structural and Functional Disruption of Neuronal Networks

Kuchibhotla, Kishore V.; Goldman, Samuel T.; Lattarulo, Carli; Wu, Hai Yan; Hyman, Bradley T.; Bacskai, Brian J.

doi:10.1016/j.neuron.2008.06.008

Cited by 572 publications

(651 citation statements)

References 55 publications

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“…Many, in fact, have used this lack of neuronal death as evidence that amyloid is not relevant to dementia in AD. We and others (1)(2)(3)(4) have identified structural and functional alterations of neurons in the brains of APP mice that implicate amyloid-mediated toxicity, but we have never detected neuronal death. The ability to monitor cell death in an experimental model provides the opportunity to intervene with neuroprotective agents that could be applied to the spectrum of neurodegenerative diseases and CNS disorders.…”

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confidence: 53%

“…The maximal ratio change using 2 mM DTDP increased the roGFP1 ratio from 1.01 ± 0.01 to 1.78 ± 0.04 (n ≥ 59 imaging sites from three mice); whereas 2 mM DTT decreased the resting roGFP ratio (0.90 ± 0.01, n ≥ 20 imaging sites from three mice). Although the dynamic range of roGFP is reduced in the living brain compared with in vitro preparations, as with other genetically encoded reporters (2), it remains a sensitive reporter of changes in redox potential. Thus, this approach provides a quantitative functional readout of intracellular oxidative stress in cortical neurons in living mouse brain.…”

Section: Resultsmentioning

confidence: 99%

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Rapid cell death is preceded by amyloid plaque-mediated oxidative stress

Xie

Hou

Jiang

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Neuronal loss is the ultimate outcome in a variety of neurodegenerative diseases and central nerve system disorders. Understanding the sequelae of events that leads to cell death would provide insight into neuroprotective approaches. We imaged neurons in the living brain of a mouse model of Alzheimer's disease that overexpresses mutant human amyloid precursor protein and presenilin 1 and followed the death of individual neurons in real time. This mouse model exhibited limited neurodegeneration and atrophy, but we were able to identify a small fraction of vulnerable cells that would not have been detectable by using standard approaches. By exploiting a genetically encoded reporter of oxidative stress, we identified susceptible neurons by their increased redox potential. The oxidative stress was most dramatic in neurites near plaques, propagated to cell bodies, and preceded activation of caspases that led to cell death within 24 h. Thus, local oxidative stress surrounding plaques contributes to longrange toxicity and selective neuronal death in Alzheimer's disease.in vivo imaging | reduction-oxidation sensitive GFP A lzheimer's disease (AD) is underscored by neurodegeneration and is the most common form of dementia. The pathological hallmarks of this disease include amyloid plaques, neurofibrillary tangles, and neuronal loss. Early-onset familial AD is caused by genetic mutations of amyloid precursor protein (APP) or presenilin 1 and 2 (PS1 and PS2). Although recent genetic studies have revealed risk factors for late onset AD, the pathogenic pathways for sporadic AD remain largely unknown. The development of mouse models of AD that develop senile plaques similar to those found in AD patients was a critical step in identifying the role of amyloid β (Aβ) on neuronal function. A major disappointment of most of the mouse models is the lack of overt neuronal loss that is a hallmark of the human disease. Many, in fact, have used this lack of neuronal death as evidence that amyloid is not relevant to dementia in AD. We and others (1-4) have identified structural and functional alterations of neurons in the brains of APP mice that implicate amyloid-mediated toxicity, but we have never detected neuronal death. The ability to monitor cell death in an experimental model provides the opportunity to intervene with neuroprotective agents that could be applied to the spectrum of neurodegenerative diseases and CNS disorders.We were able to identify vulnerable cells by quantitatively imaging the redox potential of neurons in the living brain. Our hypothesis was that amyloid-mediated increases in oxidative stress are the initiators of the toxic cascade that leads to cell loss. Accumulating evidence supports a role for oxidative stress in the pathogenesis of neuronal degeneration and death in AD (5-8). The evidence supporting oxidative stress in AD comes largely from postmortem samples and includes increased lipid peroxidation, decreased polyunsaturated fatty acids (9-12), increased protein oxidation (13,14), and DNA oxidation (15, ...

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confidence: 53%

Section: Resultsmentioning

confidence: 99%

Rapid cell death is preceded by amyloid plaque-mediated oxidative stress

Xie

Hou

Jiang

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

146

120

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“…[39][40][41][42] When Ca 2+ is measured in the spines and dendrites of cortical pyramidal neurons of transgenic mice, there was a higher than normal resting level in those neurons located close to amyloid deposits. 43 Similarly, the resting level of Ca 2+ in the cortical neurons of 3xTg-AD animals was 247 nmol/L, which was twice that found in the non-Tg controls (110 nmol/L). 44 There are indications that this increase in Ca 2+ signaling is caused by changes in both the entry of external Ca 2+ and release from internal stores.…”

Section: Bipolar Disordermentioning

confidence: 80%

“…4). 43,[45][46][47] The β-amyloid peptides, which aggregate to form complexes may enhance entry either directly by forming channels in the membrane 48 or by stimulating pre-existing channels such as the NMDARs. 49 The cellular prion protein (PrP C ), which is tethered to the outside of the membrane through a glycosyl phosphatidylinositol (GPI) anchor, functions as an amyloid β receptor and may thus carry out some of these actions on Ca 2+ entry at the plasma membrane.…”

Section: Bipolar Disordermentioning

confidence: 99%

Dysregulation of neural calcium signaling in Alzheimer disease, bipolar disorder and schizophrenia

Berridge

2013

Prion

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“…The Ca 2+ influx signals (i.e., the transmission strength) varied along the axonal branches with a gradient of weak, at the proximal boutons, to strong, at the distal ones. More recently, quantitative in vivo imaging using YC3.60 revealed elevated [Ca 2+ ] i in neurites near Aβ plaques (Kuchibhotla et al 2008). Although FRET-based indicators for Ca 2+ have been developed, our understanding of the structure-photochemistry relationships of GFP has enabled the development of Ca 2+ probes based on a single GFP variant.…”

Section: Discussionmentioning

confidence: 99%

Imaging Intracellular Free Ca²⁺ Concentration Using Yellow Cameleons

Miyawaki¹,

Nagai²,

Mizuno³

2013

Cold Spring Harb Protoc

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Green fluorescent protein (GFP)-based fluorescent indicators for Ca 2+ offer significant promise for monitoring Ca 2+ in previously unexplored organisms, tissues, and submicroscopic environments because they are genetically encoded, function without cofactors, can be targeted to any intracellular location, and are bright enough for single-cell imaging. These probes use simple GFP variants, circularly permuted GFP (cpGFP), in which the amino and carboxyl portions have been interchanged and reconnected by short spacers between the original termini, or pairs of GFP variants that permit fluorescence resonance energy transfer (FRET). Yellow cameleons (YCs) use FRET between cyanand yellow-emitting variants of Aequorea GFP (cyan fluorescent protein [CFP] and yellow fluorescent protein [YFP], respectively). YCs are composed of a linear combination of CFP, calmodulin (CaM), a glycylglycine linker, the CaM-binding peptide of myosin light-chain kinase (M13), and YFP. Binding of Ca 2+ to the CaM moiety of the YC initiates an intramolecular interaction between the CaM and the M13 domains, causing the chimeric protein to shift from an extended conformation to a more compact one, thereby increasing the efficiency of FRET from CFP to YFP. This technique is amenable to emission ratioing, which is more quantitative than single-wavelength monitoring. YC3.60 was engineered to enhance its performance as a fluorescent Ca 2+ indicator and here we describe the use of this cameleon to image rapid changes in intracellular free Ca 2+ concentration ([Ca 2+ ] i ) within HeLa cells. FRET imaging is performed using a laser-scanning confocal microscope. MATERIALSIt is essential that you consult the appropriate Material Safety Data Sheets and your institution's Environmental Health and Safety Office for proper handling of equipment and hazardous material used in this protocol. ReagentsCa 2+ -free medium For fast and simultaneous acquisition of cpVenus173 and CFP images from HeLa cells expressing YC3.60, this camera, which comprises three charge-coupled device (CCD) chips (RGB: red, green, and blue) and a prism, can be used. The cpVenus173 and CFP images are captured by the G and B chips, respectively. In addition, to improve spatial resolution along the z-axis, a spinning-disk unit (e.g., CSU21 from Yokogawa Electric Corporation) can be placed in front of the camera.Diode-pumped solid-state laser (430 nm) (Melles Griot) Glass-bottomed dishes (35 mm) (Matsunami Glass Ind., Ltd., D111300) Image acquisition software to control the Ashura 3CCD camera (e.g., . These include intensity of the laser, scanning speed, binning, and sensitivity of the camera.In our experience, the intensity of the laser should be attenuated greatly using neutral-density filters for Ca 2+ imaging on a timescale of subseconds. Photobleaching does not happen with a laser that is sufficiently attenuated.6. Observe fluorescence signals from CFP and cp173Venus by the blue and green channels of the 3CCD camera, respectively. 7. Set 4 × 4 binning for high-sensitive fluorescence ...

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Aβ Plaques Lead to Aberrant Regulation of Calcium Homeostasis In Vivo Resulting in Structural and Functional Disruption of Neuronal Networks

Cited by 572 publications

References 55 publications

Rapid cell death is preceded by amyloid plaque-mediated oxidative stress

Rapid cell death is preceded by amyloid plaque-mediated oxidative stress

Dysregulation of neural calcium signaling in Alzheimer disease, bipolar disorder and schizophrenia

Imaging Intracellular Free Ca²⁺ Concentration Using Yellow Cameleons

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Aβ Plaques Lead to Aberrant Regulation of Calcium Homeostasis In Vivo Resulting in Structural and Functional Disruption of Neuronal Networks

Cited by 572 publications

References 55 publications

Rapid cell death is preceded by amyloid plaque-mediated oxidative stress

Rapid cell death is preceded by amyloid plaque-mediated oxidative stress

Dysregulation of neural calcium signaling in Alzheimer disease, bipolar disorder and schizophrenia

Imaging Intracellular Free Ca2+ Concentration Using Yellow Cameleons

Contact Info

Product

Resources

About

Imaging Intracellular Free Ca²⁺ Concentration Using Yellow Cameleons