%0 Journal Article %T New highly sensitive rodent and human tests for soluble amyloid precursor protein alpha quantification: preclinical and clinical applications in Alzheimer's disease. %+ Centre de Psychiatrie et Neurosciences (U894) %+ Service de Biochimie et de Biologie moléculaire %+ Centre de Mémoire de Ressources et de Recherche %+ Laboratoire d'Histologie et de Biologie du Vieillissement %+ Institut du Fer à Moulin %+ Service Neuroscience %A Rose, Christiane %A Peoc'H, Katell %A Chasseigneaux, Stéphanie %A Paquet, Claire %A Dumurgier, Julien %A Bourasset, Fanchon %A Calon, Frédéric %A Laplanche, Jean-Louis %A Hugon, Jacques %A Allinquant, Bernadette %Z This work was supported by Association Internationale pour la Recherche sur la Maladie d'Alzheimer and INSERM %< avec comité de lecture %@ 1471-2202 %J BMC Neuroscience %I BioMed Central %V 13 %N 1 %P 84 %8 2012 %D 2012 %R 10.1186/1471-2202-13-84 %M 22824057 %K Alzheimer's disease %K Soluble amyloid precursor protein alpha %K Homogeneous time-resolved fluorescence %K Rodent %K Human %K Cerebrospinal fluid %K Primary neurons %K Sensitivity %Z Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]Journal articles %X UNLABELLED: ABSTRACT: BACKGROUND: Amyloid precursor protein (APP), a key molecule in Alzheimer's disease (AD), is metabolized in two alternative cleavages, generating either the amyloidogenic peptides involved in AD pathology or the soluble form of APP (sAPPα). The level of amyloidogenic peptides in human cerebrospinal fluid (CSF) is considered to be a biomarker of AD, whereas the level of sAPPα in CSF as a biomarker has not been clearly established. sAPPα has neurotrophic and neuroprotective properties. Stimulating its formation and secretion is a promising therapeutic target in AD research. To this end, very sensitive tests for preclinical and clinical research are required. METHODS: The tests are based on homogenous time-resolved fluorescence and require no washing steps. RESULTS: We describe two new rapid and sensitive tests for quantifying mouse and human sAPPα. These 20 μl-volume tests quantify the levels of: i) endogenous mouse sAPPα in the conditioned medium of mouse neuron primary cultures, as well as in the CSF of wild-type mice, ii) human sAPPα in the CSF of AD mouse models, and iii) human sAPPα in the CSF of AD and non-AD patients. These tests require only 5 μl of conditioned medium from 5 × 104 mouse primary neurons, 1 μl of CSF from wild-type and transgenic mice, and 0.5 μl of human CSF. CONCLUSIONS: The high sensitivity of the mouse sAPPα test will allow high-throughput investigations of molecules capable of increasing the secretion of endogenous sAPPα in primary neurons, as well as the in vivo validation of molecules of interest through the quantification of sAPPα in the CSF of treated wild-type mice. Active molecules could then be tested in the AD mouse models by quantifying human sAPPα in the CSF through the progression of the disease. Finally, the human sAPPα test could strengthen the biological diagnosis of AD in large clinical investigations. Taken together, these new tests have a wide field of applications in preclinical and clinical studies. %G English %2 https://inserm.hal.science/inserm-00723764/document %2 https://inserm.hal.science/inserm-00723764/file/1471-2202-13-84.pdf %L inserm-00723764 %U https://inserm.hal.science/inserm-00723764 %~ INSERM %~ UNIV-PARIS5 %~ UNIV-PARIS7 %~ UPMC %~ APHP %~ IFM %~ UPMC_POLE_4 %~ SORBONNE-UNIVERSITE %~ SU-SCIENCES %~ UNIV-PARIS %~ UP-SANTE %~ SU-TI %~ ALLIANCE-SU