Altered amyloid precursor protein (APP) processing leading to increased production and oligomerization of A␤ may contribute to Alzheimer's disease (AD). Understanding how APP processing is regulated under physiological conditions may provide new insights into AD pathogenesis. Recent reports demonstrate that excitatory neural activity regulates APP metabolism and A␤ levels, although understanding of the molecular mechanisms involved is incomplete. We have investigated whether NMDA receptor activity regulates APP metabolism in primary cultured cortical neurons. We report that a pool of APP is localized to the postsynaptic compartment in cortical neurons and observed partial overlap of APP with both NR1 and PSD-95. NMDA receptor stimulation increased nonamyloidogenic ␣-secretase-mediated APP processing, as measured by a 2.5-fold increase in cellular ␣-C-terminal fragment (C83) levels after glutamate or NMDA treatment. This increase was blocked by the NMDA receptor antagonists D-AP5 and MK801 but not by the AMPA receptor antagonist CNQX or the L-type calcium channel blocker nifedipine, was prevented by chelation of extracellular calcium, and was blocked by the ␣-secretase inhibitor TAPI-1. Cotreatment of cortical neurons with bicuculline and 4-AP, which stimulates glutamate release and activates synaptic NMDA receptors, evoked an MK801-sensitive increase in C83 levels. Furthermore, NMDA receptor stimulation caused a twofold increase in the amount of soluble APP detected in the neuronal culture medium. Finally, NMDA receptor activity inhibited both A␤1-40 release and Gal4-dependent luciferase activity induced by ␤-␥-secretase-mediated cleavage of an APP-Gal4 fusion protein. Altogether, these data suggest that calcium influx through synaptic NMDA receptors promotes nonamyloidogenic ␣-secretase-mediated APP processing.

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This is a study of the interaction between the two NMDA neurotransmitter receptor subtypes, NR1/NR2A and NR1/NR2B, and amyloid precursor protein (APP) 695, the major APP variant expressed in neurones. APP695 co‐immunoprecipitated with assembled NR1‐1a/NR2A and NR1‐1a/NR2B NMDA receptors following expression in mammalian cells. Single NR1‐1a, NR1‐2a, NR1‐4bc‐Myc, or NR2 subunit transfections revealed that co‐association of APP695 with assembled NMDA receptors was mediated via the NR1 subunit; it was independent of the NR1 C1, C2, and C2′ cassettes and, the use of an NR1‐2ac‐Myc‐trafficking mutant suggested that interaction between the two proteins occurs in the endoplasmic reticulum. The use of antibodies directed against extracellular and intracellular NR2 subunit epitopes for immunoprecipitations suggested that APP/NMDA receptor association was mediated via N‐terminal domains. Anti‐APP antibodies immunoprecipitated NR1, NR2A, and NR2B immunoreactive bands from detergent extracts of mammalian brain; reciprocally, anti‐NR1 or anti‐NR2A antibodies co‐immunoprecipitated APP immunoreactivity. Immune pellets from brain were sensitive to endoglycosidase H suggesting that, as for heterologous expression, APP and NMDA receptor association occurs in the endoplasmic reticulum. Co‐expression of APP695 in mammalian cells resulted in enhanced cell surface expression of both NR1‐1a/NR2A and NR1‐1a/NR2B NMDA receptors with no increase in total subunit expression. These findings are further evidence for a role of APP in intracellular trafficking mechanisms. Further, they provide a link between two major brain proteins that have both been implicated in Alzheimer’s disease.

Abdominal computed tomography (CT) studies of 19 dogs with no history or clinical signs of gastrointestinal disease, and two dogs with a histological diagnosis of gastrointestinal neoplasia were examined retrospectively. Gastrointestinal segments were evaluated subjectively for conspicuity, contrast enhancement, and wall layering after contrast medium administration. In dogs without gastrointestinal disease, there were 62.8% of gastrointestinal segments (serosa to serosa) and 77.7% of gastrointestinal walls (serosa to mucosa) visualized. Wall layering on postcontrast images was seen in 21.8% of gastrointestinal segments. There was significant association between gastrointestinal diameter and wall thickness. There was significant association between weight and gastrointestinal wall thickness in the following regions: gastric fundus, gastric body, gastric pylorus, gastric pyloric antrum, duodenal cranial flexure, jejunum and ascending colon, and between patient weight and gastrointestinal diameter in cranial duodenal flexure, descending duodenum, transverse duodenum, ascending duodenum, and jejunum. Measurements acquired from CT studies correlated well with previously published normal reference ranges for radiographic and ultrasonographic studies. Gastrointestinal neoplasia, diagnosed in two dogs, had a gastrointestinal wall thickness greater than the range of the dogs without gastrointestinal disease. Computed tomography offers identification of the gastrointestinal tract segments in dogs, allows for evaluation of gastrointestinal diameter and aids in investigation of gastrointestinal wall thickness.

There was a small side-to-side variation of radial torsion in normal dogs. With directed training, torsion assessment using CT is reliable in dogs with and without antebrachial bone deformity.