Poster Presentations: P1 P181

with APP695 by Co-Immunoprecipitation with HSP56, HSP70, and

UBQLN1 antibodies suggesting that these chaperones are associated with

APP695. Conclusions: In summary, we conclude that HSP56, HSP70,

and UBQLN1 may play a direct role in the in the folding of APP695 and

may be useful in therapeutic uses for degradation of Amyloid-b.

P1-079 BETA-AMYLOID–INDUCED NEUROTOXICITY

MIGHT BE REDUCED THROUGH AMPK-

MEDIATED BY DONEPEZIL

Moon Ho Park1, Kun-Woo Park2, Hyeon Soo Kim3, 1Korea University

Medical College, Seoul, South Korea; 2Korea University Medical College,

Seoul, South Korea; 3Korea University, Seoul, South Korea.

Contact e-mail: parkmuno@yahoo.co.kr

Background: Acetylcholinesterase inhibitors (AchEIs) and NMDA recep-

tor anatagonist memantine are used for the treatment of Alzheimer’s dis-

ease. Recently, the AchEI donepezil and memantine were found to have

neuroprotective effects. However, these protective mechanisms have not

yet been clearly identified. AMP-activated protein kinase (AMPK), a master

regulator of cellular energy homeostasis and a central player in glucose and

lipid metabolism, is potentially implicated in the pathogensis of Alzheim-

er’s disease. We investigated the neuroprotective effects of donepezil and

memantine against amyloid-b 1-42 (Ab42)-induced neurotoxicity and

AMPK signal pathway in HN33 hippocampal cell line.Methods: The neu-

roprotective effects of treatment with donepezil andmemantine on Ab42-in-

duced cell death, was measured by MTT assay. Expression levels of

phosphorylation status of AMPK were analyzed by Western blot. Intracel-

lular calcium concentration was measured with calcium indicator dye,

fluo-3AM. Results:We reported that only donepezil increased intracellular

calcium concentration and donepezil activated AMPK in HN33 cells with

Ab42. Moreover, inhibition of AMPK resulted in blockage of donepezil-in-

duced calcium uptake. Conclusions: These results suggest that donepezil

might prevent Ab42-induced neurotoxicity through AMPK pathway.

P1-080 TIME-COURSE OF BEHAVIORAL AND

L-ARGININEMETABOLIC PROFILE CHANGES IN

THE APP SWE PS1 DE9 MOUSE MODEL OF

ALZHEIMER’S DISEASE

David Bergin1, Yu Jing1, Hu Zhang2, BruceMockett3,Wickliffe Abraham1,

Ping Liu1, 1University of Otago, Dunedin, New Zealand; 2School of

Pharmacy, University of Otago, Dunedin, New Zealand; 3Department of

Psychology, University of Otago, Dunedin, New Zealand.

Contact e-mail: davebergin@anatomy.otago.ac.nz

Background: Alzheimer’s disease (AD) is a common neurodegenerative

disease among the elderly. Due to complicated idiopathic pathology, AD

is best characterized by progressive cognitive decline with memory loss

as the earliest prognostic indicator. Mounting evidence indicates that L-ar-

ginine metabolism - integral for cell homeostasis, growth and memory - is

altered in AD. However L-arginine metabolism has not been investigated

in transgenic (Tg) animal models that are used to mimic and understand

pathological AD processes. Methods: The current study examined the be-

havioral and L-arginine metabolic profile in 7 and 13 month old male

APP SWE PS1 DE9 mice using both reference and working memory ver-

sions of the Morris water maze task, high-performance liquid chromatogra-

phy and liquid chromatography mass spectroscopy. Results:We found that

7 month old Tg mice demonstrated mild impairments in spatial working

memory, increased agmatine in the parahippocampal cortex, and increased

spermine concentrations in the frontal cortex and striatum compared to their

wild-type littermates. At 13 months of age, Tg mice were significantly im-

paired in both versions of the watermaze task and had accompanying alter-

ations in L-arginine, L-ornithine, agmatine, putrescine, spermidine or

spermine in at least one of the five brain regions examined. There were

no genotype-specific differences in the cerebellum at either age. Nor were

there genotype specific differences in glutamate or GABA at either age

points in any brain region. Conclusions: These results indicate that L-argi-

nine metabolism is increasingly altered with age in the APP SWE PS1 DE9

mice, which reflect and may influence the behavioral impairments observed.

P1-081 IDENTIFICATION OFAN ALTERNATIVE APP

PROCESSING PATHWAY

Michael Willem1, Sabina Tahirovic2, Ignasi Forn�e3, Elisabeth Kremmer4,

Christiane Volbracht5, H�el�ene Marie6, Christian Haass7, 1University

Munich, Munich, Germany; 2DZNE-M€unchen, Munich, Germany;3Adolf-Butenandt-Institute, Munich, Germany; 4Hemholtz-Center Munich,

Munich, Germany; 5Lundbeck, Copenhagen, Denmark; 6IPMC-CNRS

UMR7275, Sophia-Antipolis, France; 7DZNE & LMU Munich, Munich,

Germany. Contact e-mail: mwillem@med.uni-muenchen.de

Background:Generation of amyloid b -peptide (Ab) from the amyloid pre-

cursor protein (APP) depends on sequential cleavage by b - and b -secretase.

The aspartylprotease BACE1 (b -site APP cleaving enzyme-1) is known to

be the sole b-secretase in the brain, based on the complete absence of b-sec-

retase cleavage products in BACE1 -/- mice and massively reduced Ab

levels in mouse brains upon treatment with a BACE1 inhibitor. Methods:

We investigated APP processing in the absence of BACE1 activity upon

pharmacological inhibition and siRNA knock down in CHO cells and pri-

mary neurons. For in vivo analysis, we used brain lysates of BACE1 KO

mice and inhibitor treated animals. Epitope specific monoclonal antibodies

were produced and applied for biochemical identification of APP processing

products, e.g. by SEC and LC-MS/MS analysis.Results: Biochemical anal-

ysis revealed that pharmacological blockade of b-secretase, results not only

in reduction of A b synthesis but also in accumulation of novel APP-C-ter-

minal fragments. The fragments generated in the absence of BACE1 are N-

terminally extended, which could be demonstrated with specific antibodies

directed against this region.We identified and characterized a corresponding

secreted fragment in supernatants of APP expressing. A similar cleavage

pattern is observed in primary neuronal cultures and in mouse brain where

it is enriched upon BACE1 blockade. Conclusions: With these studies we

identified new APP cleavage products which could potentially influence

not only A b production but also Ab toxicity.

P1-082 TYROSINE KINASE INHIBITION PROMOTES

PARKIN-BECLIN-1–MEDIATED PROTECTION IN

ALZHEIMER’S DISEASE MODELS

Irina Lonskaya, Michaeline Hebron, Charbel Moussa, Georgetown

University, Washington, D.C., United States. Contact e-mail: cem46@

georgetown.edu

Background: The tyrosine kinase c-Abl is activated in neurodegenerative

disorders, including Alzheimer’s disease (AD). Nilotinib is a c-Abl inhibitor

approved by the U.S. Food and Drug Administration (FDA) for treatment of

adult leukemia.We evaluated the effects of Nilotinib on c-Abl inhibition and

autophagic clearance of b-amyloid and Tau in models of AD.Methods:We

injected mice IP with 10mg/kg Nilotinib once a day for 3consecutive weeks

and performed several biochemical, histological and behavioral tests to

evaluate Nilotinib effects on amyoid clearance and cognition. Results: Pe-

ripheral administration of Nilotinib inhibited c-Abl and other tyrosine kin-

asesand resulted in activation of the E3 ubiquitin ligase parkin, and elevation

of beclin-1 in AD mice brain. Nilotinib-mediated parkin activation stimu-

lated the autophagic clearance pathway, leading to amyloid degradation

and cognitive improvement in a parkin-dependent manner. Nilotinib failed

to clear autophagic vacuoles and amyloid proteins in parkin -/- mice, despite

an increase in beclin-1 levels, whereas beclin-1 knockdown attenuated A

b clearance, underscoring an indispensable role for endogenous parkin in

autophagy. These data suggest that Nilotinib-mediated c-Abl inhibition is

a therapeutic strategy to rescue cells from intraneuronal amyloid toxicity

and prevent both plaque deposition and progression frommild cognitive im-

pairment to AD.Conclusions:Although no parkin mutations are associated

with AD, parkin manipulation may be a disease modifying therapy that pro-

vides an alternative approach to prevent progression from MCI to AD. Ni-

lotinib is FDA approved, re-purposing its use is not only useful for MCI

treatment but were it to be effective, it also could be used to test the validity

of disease biomarkers such as p-Tau and b -amyloid. These studies provide