Imaging High-Resolution Structure of GFP-Expressing Neurons in Neocortex In Vivo

Chen, Brian E.; Lendvai, Balázs; Nimchinsky, Esther A.; Burbach, Barry; Fox, Kevin; Svoboda, Karel

doi:10.1101/lm.32700

Cited by 61 publications

(43 citation statements)

References 61 publications

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“…The cranial-window surgical procedure was performed as described elsewhere with slight modifications (23,35). Briefly, animals were anesthetized with isoflurane (4% induction, 1.5% maintenance) while being immobilized in a custom-made animal stereotaxic apparatus equipped with an anesthetic mask.…”

Section: Methodsmentioning

confidence: 99%

Anti-Aβ antibody treatment promotes the rapid recovery of amyloid-associated neuritic dystrophy in PDAPP transgenic mice

Brendza¹,

Bacskai²,

Cirrito³

et al. 2005

J. Clin. Invest.

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Neuritic plaques are a defining feature of Alzheimer disease (AD) pathology. These structures are composed of extracellular accumulations of amyloid-β peptide (Aβ) and other plaque-associated proteins, surrounded by large, swollen axons and dendrites (dystrophic neurites) and activated glia. Dystrophic neurites are thought to disrupt neuronal function, but whether this damage is static, dynamic, or reversible is unknown. To address this, we monitored neuritic plaques in the brains of living PDAPP;Thy-1:YFP transgenic mice, a model that develops AD-like pathology and also stably expresses yellow fluorescent protein (YFP) in a subset of neurons in the brain. Using multiphoton microscopy, we observed and monitored amyloid through cranial windows in PDAPP;Thy-1:YFP double-transgenic mice using the in vivo amyloid-imaging fluorophore methoxy-X04, and individual YFP-labeled dystrophic neurites by their inherent fluorescence. In vivo studies using this system suggest that amyloid-associated dystrophic neurites are relatively stable structures in PDAPP;Thy-1:YFP transgenic mice over several days. However, a significant reduction in the number and size of dystrophic neurites was seen 3 days after Aβ deposits were cleared by anti-Aβ antibody treatment. This analysis suggests that ongoing axonal and dendritic damage is secondary to Aβ and is, in part, rapidly reversible. IntroductionAlzheimer disease (AD) is a neurodegenerative disorder that results in memory deficits, changes in personality, and cognitive decline. It is the leading cause of dementia in the US, affecting approximately 10% of those over 65 and 50% of those over 85 years of age. One of the invariant pathological hallmarks of AD is the presence of neuritic plaques in areas of the brain responsible for memory and cognition. Neuritic plaques consist predominantly of extracellular fibrils of amyloid-β peptide (Aβ) and are closely associated with dystrophic neurites, activated microglia, and reactive astrocytes (1-3). The actual mechanisms that contribute to the pathogenesis of AD are not known; however, compelling genetic and biochemical evidence suggests that accumulation of amyloid-β protein plays a central role. Thus, preventing or reversing the formation of amyloid may be a viable treatment.The dystrophic neurites that surround amyloid deposits are markedly swollen, distorted axons and dendrites. In AD, the number of dystrophic neurites has been shown to correlate with the clinical severity of dementia (4), and neuronal dystrophy is associated with synaptic loss in cortical cultures exposed to fibrillar Aβ (5). Studies in human AD and in transgenic models suggest that alterations in dendritic curvature and morphology, including neuritic dystrophy, that are associated with deposits of fibrillar Aβ

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Section: Methodsmentioning

confidence: 99%

Anti-Aβ antibody treatment promotes the rapid recovery of amyloid-associated neuritic dystrophy in PDAPP transgenic mice

Brendza¹,

Bacskai²,

Cirrito³

et al. 2005

J. Clin. Invest.

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“…Multiphoton microscopy has been previously used in intravital studies of morphology and physiology of the brain [3, 4, 5, 6, 7, 8], metabolism of the skin [9], development and vascularization of tumors [10], and embryonic development [11]. Recently, the extended optical reach provided by multiphoton microscopy was applied to imaging of the kidneys of living rats [12].…”

Section: Multiphoton Microscopy and Intravital Microscopymentioning

confidence: 99%

Intravital Imaging of the Kidney Using Multiparameter Multiphoton Microscopy

Dunn¹,

Sandoval²,

Molitoris

2004

Nephron Exp Nephrol

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Intravital optical microscopy provides a powerful means of studying the cell biology in the most physiologically relevant setting. The ability of multiphoton microscopy to collect optical sections deep into biological tissues has opened up the field of intravital microscopy to high-resolution studies of multiple organs. Presented here are examples of how two-photon microscopy can be applied to intravital studies of kidney physiology and the study of disease processes. These include studies of cell vitality and apoptosis, fluid transport, receptor-mediated endocytosis, blood flow, and leukocyte trafficking. Efficient two-photon excitation of multiple fluorophores permits comparison of multiple probes and simultaneous characterization of multiple parameters. Two-photon microscopy can now provide a level of investigation previously unattainable in intravital microscopy, enabling kinetic analyses and physiological studies of the organs of living animals with subcellular resolution. Therefore, application of this technology will provide direct visualization of organ-specific and cell-specific responses to an array of stimuli and therapeutic approaches, enhancing our understanding and treatment of disease processes.

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“…A small window was opened in the skull 2 mm posterior and 4.5 mm lateral to the anterior fontanel. These coordinates reliably target the barrel cortex in neonatal rats (Chen et al, 2000a;Lendvai et al, 2000;Takahashi et al, 2003). Glass pipettes (tip diameter, ϳ10 -12 m) were used to penetrate the dura and to pressure-inject (ϳ15 psi) recombinant Sindbis virus into the barrel cortex while avoiding damage to surface blood vessels.…”

Section: Gfp-tagged Constructs and Expression Strategiesmentioning

confidence: 99%

Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity

Ehrlich¹,

Malinow²

2004

J. Neurosci.

492

413

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The regulated delivery of AMPA-type glutamate receptors (AMPARs) to synapses is an important mechanism underlying synaptic plasticity. Here, we ask whether the synaptic scaffolding protein PSD-95 (postsynaptic density 95) participates in AMPAR incorporation during two forms of synaptic plasticity. In hippocampal slice cultures, the expression of PSD-95-green fluorescent protein (PSD-95-GFP) increases AMPAR currents by selectively delivering glutamate receptor 1 (GluR1)-containing receptors to synapses, thus mimicking long-term potentiation (LTP). Mutational analysis shows that the N terminal of PSD-95 including the first two PDZ [ PSD-95/Discs large (Dlg)/zona occludens-1 (ZO-1)] domains is necessary and sufficient to mediate this effect. Further supporting a role in synaptic plasticity, wild-type PSD-95 occludes LTP and dominant negative forms block LTP. Moreover, we demonstrate that PSD-95 also participates in AMPAR delivery during experience-driven plasticity in vivo. In the barrel cortex from experience-deprived animals, the expression of PSD-95-GFP selectively increases AMPAR currents, mimicking experience-driven plasticity. In nondeprived animals, PSD-95-GFP produces no additional potentiation, indicating common mechanisms between PSD-95-mediated potentiation and experience-driven synaptic strengthening. A dominant negative form of PSD-95 blocks experience-driven potentiation of synapses. Pharmacological analysis in slice cultures reveals that PSD-95 acts downstream of other signaling pathways involved in LTP. We conclude that PSD-95 controls activity-dependent AMPAR incorporation at synapses via PDZ interactions not only during LTP in vitro but also during experience-driven synaptic strengthening by natural stimuli in vivo.

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Imaging High-Resolution Structure of GFP-Expressing Neurons in Neocortex In Vivo

Cited by 61 publications

References 61 publications

Anti-Aβ antibody treatment promotes the rapid recovery of amyloid-associated neuritic dystrophy in PDAPP transgenic mice

Anti-Aβ antibody treatment promotes the rapid recovery of amyloid-associated neuritic dystrophy in PDAPP transgenic mice

Intravital Imaging of the Kidney Using Multiparameter Multiphoton Microscopy

Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity

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