The processing and biological function of the human amyloid precursor protein (APP): lessons from different cellular models.

One of the major neuropathological hallmarks of Alzheimer's disease is the presence of senile plaques in vulnerable regions of CNS. These plaques are formed of aggregated amyloid peptide. Amyloid peptide is released by the cleavage of its precursor (APP). The establishment of cell lines expressing human APP allowed to characterize both amyloidogenic and non-amyloidogneic pathways of APP catabolism and to identify some of the proteins involved in this processing (known as secretases). This led to a better comprehension of amyloid peptide production, which needs to be further characterized since gamma-secretase is as yet not identified; moreover, we still lack a clear overview of the interactions between APP and other proteins promoting Alzheimer's disease (tau, presinilinsellipsis). An important limitation of these cell lines for studying the mechanisms involved in Alzheimer's disease is supported by the observation that human APP expression does not modify transfected cells survival. The infection of primary neuronal cultures with full-length human APP indicates that APP expression induces neuronal apoptosis by itself; this neurotoxicity does not rely on extracellular production of APP derivatives (secreted APP, amyloid peptide). It is now essential to understand, in neuronal models, the production, localization and involvement of amyloid peptide in neurodegenerative processes

Intracellular amyloid-beta 1-42, but not extracellular soluble amyloid-beta peptides, induces neuronal apoptosis.

Alzheimer disease (AD), the most frequent cause of dementia, is characterized by an important neuronal loss. A typical histological hallmark of AD is the extracellular deposition of beta-amyloid peptide (A beta), which is produced by the cleavage of the amyloid precursor protein (APP). Most of the gene mutations that segregate with the inherited forms of AD result in increasing the ratio of A beta 42/A beta 40 production. A beta 42 also accumulates in neurons of AD patients. Altogether, these data strongly suggest that the neuronal production of A beta 42 is a critical event in AD, but the intraneuronal A beta 42 toxicity has never been demonstrated. Here, we report that the long term expression of human APP in rat cortical neurons induces apoptosis. Although APP processing leads to production of extracellular A beta 1-40 and soluble APP, these extracellular derivatives do not induce neuronal death. On the contrary, neurons undergo apoptosis as soon as they accumulate intracellular A beta 1-42 following the expression of full-length APP or a C-terminal deleted APP isoform. The inhibition of intraneuronal A beta 1-42 production by a functional gamma-secretase inhibitor increases neuronal survival. Therefore, the accumulation of intraneuronal A beta 1-42 is the key event in the neurodegenerative process that we observed

Inhibition of neuronal calcium oscillations by cell surface APP phosphorylated on T668.

Adenoviral expression of human APP (hAPP), but not of hAPP deleted from its C-terminal intracellular domain, in rat cortical neurons abolishes spontaneous synchronous calcium oscillations. The intracellular domain of APP695 contains several residues that can be phosphorylated. Contrary to non-neuronal cells, a very high phosphorylation of APP on T668 is observed in neurons, which is mediated by JNK, GSK3 and Cdk5 protein kinases. JNK activity, modulated by GSK3, enhances the traffic of phosphorylated APP to nerve terminals, contrary to Cdk5. Here we show that inhibition of GSK3 and JNK restores calcium oscillations in an hAPP expressing neuronal network, whereas inhibition of Cdk5 does not. Expression of mutant hAPPT668A does not inhibit calcium oscillations, and the proportion of hAPPT668A at the plasma membrane is reduced by more than 50%. Altogether, these results indicate that the intracellular domain of APP is needed to inhibit neuronal calcium oscillations because GSK3/JNK phosphorylation of T668 controls APP trafficking at the plasma membrane

The role of presenilin-1 in the gamma-secretase cleavage of the amyloid precursor protein of Alzheimer's disease.

Presenilin-1 (PS1) is required for the release of the intracellular domain of Notch from the plasma membrane as well as for the cleavage of the amyloid precursor protein (APP) at the gamma-secretase cleavage site. It remains to be demonstrated whether PS1 acts by facilitating the activity of the protease concerned or is the protease itself. PS1 could have a gamma-secretase activity by itself or could traffic APP and Notch to the appropriate cellular compartment for processing. Human APP 695 and PS1 were coexpressed in Sf9 insect cells, in which endogenous gamma-secretase activity is not detected. In baculovirus-infected Sf9 cells, PS1 undergoes endoproteolysis and interacts with APP. However, PS1 does not cleave APP in Sf9 cells. In CHO cells, endocytosis of APP is required for Abeta secretion. Deletion of the cytoplasmic sequence of APP (APPDeltaC) inhibits both APP endocytosis and Abeta production. When APPDeltaC and PS1 are coexpressed in CHO cells, Abeta is secreted without endocytosis of APP. Taken together, these results conclusively show that, although PS1 does not cleave APP in Sf9 cells, PS1 allows the secretion of Abeta without endocytosis of APP by CHO cells

Search for massive neutrinos in muon-capture: development of a gas scintillation detector

A measurement of the triton recoil-spectrum from the reaction mu /sup -/+/sup 3/He+/sup 3/H+ nu /sub mu / should allow the observation of possible heavy neutrinos in the mass range 30-90 MeV. The ideal tool for the determination of the kinetic energy of the triton in a gas scintillation proportional chamber (GSPC). Two problems have to be solved in building a Helium-GSPC. The first one is that there is a mismatch of the emission spectrum (600-1000 A ) with the sensitive region of the photomultiplier. The goal of these tests is to show that small quantities of nitrogen (100-5000 p.p.m.) added to the gas can replace the normally used wavelength shifters. Secondly, with its quenching property, nitrogen should prevent the UV-light to reach the walls of the counter where these hard photons from He/sub 2//sup +/ can cause photoeffect and liberate new electrons.Anglai

Optical-model Description of Alpha+o-16 Elastic-scattering and Alpha-cluster Structure in Ne-20

O16(,) elastic scattering angular distributions have been measured for incident energies 39.3, 49.5, and 69.5 MeV. These data, and previous measurements at 32.2, 104, and 146 MeV, have been subjected to a global optical model analysis. The deduced global potential has two energy-dependent parameters which are found to vary smoothly with energy and it is uniquely determined by the data. Backward angular distributions measured between 40 and 54 MeV are also presented and shown to be nicely reproduced by the model. The sensitivity of the cross sections to the various regions of the real potential has been investigated as a function of energy using the notch test technique. The low energy behavior of the differential cross sections can be understood in terms of the semiclassical decomposition of Brink and Takigawa. A natural extrapolation of the global potential below 30 MeV is shown to reproduce the wide bump observed in the experimental excitation functions around 20 MeV. This bump is shown to be due to an l=8 shape resonance and is interpreted as the J=8+ member of the K=04+ rotational band of Ne20, in contradiction with the current attribution. Other bound and quasibound states supported by the potential are discussed in the light of orthogonality condition model-type arguments and shown to be consistent with the well-known K=01+ and 0- bands, and with the first three states of the K=04+ band of Ne20. NUCLEAR REACTIONS O16(,), measured (), E=39.3, 49.5, 69.5 MeV; global optical model analysis, E=32-146 MeV; semiclassical decomposition of the scattering amplitude; investigation of the compatibility of the potential description with existing low energy data and comparison with cluster models. © 1983 The American Physical Society.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Epigenetic regulations of immediate early genes expression involved in memory formation by the amyloid precursor protein of Alzheimer disease

We previously demonstrated that APP epigenetically regulates Egr1 expression both in cultured neurons and in vivo. Since Egr1 is an immediate early gene involved in memory formation, we wondered whether other early genes involved in memory were regulated by APP and we studied molecular mechanisms involved. By comparing prefrontal (PF) cortex from wild type (APP+/+) and APP knockout mice (APP-/-), we observed that APP down regulates expression of four immediate early genes, Egr1, c-Fos, Bdnf and Arc. Down regulation of Egr1, c-Fos and Bdnf transcription resulted from a decreased enrichment of acetylated histone H4 on the corresponding gene promoter. Further characterization of H4 acetylation at Egr1 and c-Fos promoters revealed increased acetylation of H4K5 and H4K12 residues in APP-/- mice. Whereas APP affected Egr1 promoter activity by reducing access of the CREB transcription factor, its effect on c-Fos appeared to depend on increased recruitment of HDAC2 histone deacetylase to the gene promoter. The physiological relevance of the epigenetic regulation of Egr1 and c-Fos gene transcription by APP was further analyzed following exposure of mice to novelty. Although transcription of Egr1 and c-Fos was increased following exposure of APP+/+ mice to novelty, such an induction was not possible in APP-/- mice with a high basal level of expression of these immediate early genes. Altogether, these results demonstrate that APP-mediated regulation of c-Fos and Egr1 by different epigenetic mechanisms is needed for their induction during exposure to novelty

Search for heavy neutrinos in the capture of muons by /sup 3/He: development of a helium-filled gas scintillation proportional counter

The authors have developed a high-pressure /sup 3/He target based on the principle of a gas scintillation proportional counter (GSPC) to observe with high luminosity and good energy resolution tritons emitted in the capture of muons by /sup 3/He. The techniques used with this new detector are described and the first results obtained in the search for massive neutrinos which couple to muons are shown.Anglai