Senile plaques accumulate over the course of decades in the brains of patients with Alzheimer's disease. A fundamental tenet of the amyloid hypothesis of Alzheimer's disease is that the deposition of amyloid-β precedes and induces the neuronal abnormalities that underlie dementia 1 . This idea has been challenged, however, by the suggestion that alterations in axonal trafficking and morphological abnormalities precede and lead to senile plaques 2 . The role of microglia in accelerating or retarding these processes has been uncertain. To investigate the temporal relation between plaque formation and the changes in local neuritic architecture, we used longitudinal in vivo multiphoton microscopy to sequentially image young APPswe/PS1d9xYFP (B6C3-YFP) transgenic mice 3 . Here we show that plaques form extraordinarily quickly, over 24 h. Within 1-2 days of a new plaque's appearance, microglia are activated and recruited to the site. Progressive neuritic changes ensue, leading to increasingly dysmorphic neurites over the next days to weeks. These data establish plaques as a critical mediator of neuritic pathology.To explore the formation of amyloid plaques and to determine the effects of newly formed dense-cored plaques on the microarchitecture of the brain, we have developed a novel in vivo multiphoton imaging technique. This recognizes newly formed plaques and allows monitoring of their immediate vicinity thereafter to determine the rate of their formation and the temporal sequence of pathophysiological events. We imaged young (5-to 6-month-old) B6C3-YFP mice, an age when plaques begin to appear 4 (Fig. 1). We used three-colour imaging to establish fiduciary markers for repeated imaging: YFP positive neurons, dendrites and axons in the cortex, methoxy-XO4-labelled fibrillar amyloid-β deposits in the parenchyma and on vessel walls, and a fluorescent angiogram with Texas red dextran to image blood vessels. A volume of cortex (lamina I-III) that initially did not contain plaques was re-imaged until repeat imaging detected a new plaque, establishing its 'birthday'. To ensure that the appearance of a new plaque did not simply reflect a greater depth of imaging or a slightly different imaging volume, we went through each image stack and compared NIH Public Access Author ManuscriptNature. Author manuscript; available in PMC 2012 January 23. We postulated that we would occasionally observe the appearance and growth of new plaques within an imaging volume if the time interval between imaging sessions was long enough. From one weekly imaging session to the next, most of the sites remained unchanged ( Supplementary Fig. 1a-c). However, we identified 14 new plaques: instances in which a plaque appeared in a second imaging session in a volume that had clearly been unoccupied in the first images one week earlier (Fig. 1a-c).We examined the spatial relation between newly identified plaques and blood vessels. Measurements of the distance between vessel wall and the edge of a plaque confirmed that dense-core plaques develop...

Synapse loss correlates with a cognitive decline in Alzheimer's disease (AD), but whether this is caused by fibrillar deposits known as senile plaques or soluble oligomeric forms of amyloid ␤ (A␤) is controversial. By using array tomography, a technique that combines ultrathin sectioning of tissue with immunofluorescence, allowing precise quantification of small structures, such as synapses, we have tested the hypothesis that oligomeric A␤ surrounding plaques contributes to synapse loss in a mouse model of AD. We find that senile plaques are surrounded by a halo of oligomeric A␤. Analysis of >14,000 synapses (represented by PSD95-stained excitatory synapses) shows that there is a 60% loss of excitatory synapses in the halo of oligomeric A␤ surrounding plaques and that the density increases to reach almost control levels in volumes further than 50 m from a plaque in an approximately linear fashion (linear regression, r 2 ‫؍‬ 0.9; P < 0.0001). Further, in transgenic cortex, microdeposits of oligomeric A␤ associate with a subset of excitatory synapses, which are significantly smaller than those not in contact with oligomeric A␤. The proportion of excitatory synapses associated with A␤ correlates with decreasing density (correlation, ؊0.588; P < 0.0001). These data show that senile plaques are a potential reservoir of oligomeric A␤, which colocalizes with the postsynaptic density and is associated with spine collapse, reconciling the apparently competing schools of thought of ''plaque'' vs. ''oligomeric A␤'' as the synaptotoxic species in the brain of AD patients.Alzheimer ͉ array tomography ͉ neurodegeneration ͉ synaptotoxicity L oss of connectivity caused by neuronal death and synapse loss is thought to underlie cognitive decline in neurodegenerative conditions, such as Alzheimer's disease (AD). Synapse loss appears to be particularly important in the pathogenesis of AD. Indeed, it is known that synapses are lost during AD and that in AD tissue, synapse loss correlates strongly with cognitive decline (1-3). There is a growing consensus, based primarily on cell-based assays, that amyloid ␤ (A␤), the main component of senile plaques, is toxic to synapses (4-6). In both AD patients and animal models of the disease, synapse loss is greatest near senile plaques, indicating a link between amyloid pathology and synaptotoxicity in vivo. Work by several groups has shown a decrease in dendritic spine density and synaptophysin-positive synapses radiating out from the surface of plaques in mouse models of AD (7-10). Whether this is caused by fibrillar plaques or soluble oligomeric A␤ is controversial. We used multiphoton imaging of the living brain to show that this spine loss is caused by impaired spine stability over time near plaques and postulated that a plaque-related diffusible bioactive molecule was responsible (11). Here, we test the hypothesis that oligomeric A␤ is directly synaptotoxic.We hypothesize that soluble oligomeric A␤ associates with the postsynaptic density and causes the loss of synapses and spines observ...

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