Alzheimer's Disease-Linked Mutations in Presenilin-1 Result in a Drastic Loss of Activity in Purified γ-Secretase Complexes

BACKGROUND: Mutations linked to early onset, familial forms of Alzheimer's disease (FAD) are found most frequently in PSEN1, the gene encoding presenilin-1 (PS1). Together with nicastrin (NCT), anterior pharynx-defective protein 1 (APH1), and presenilin enhancer 2 (PEN2), the catalytic subunit PS1 constitutes the core of the γ-secretase complex and contributes to the proteolysis of the amyloid precursor protein (APP) into amyloid-beta (Aβ) peptides. Although there is a growing consensus that FAD-linked PS1 mutations affect Aβ production by enhancing the Aβ1-42/Aβ1-40 ratio, it remains unclear whether and how they affect the generation of APP intracellular domain (AICD). Moreover, controversy exists as to how PS1 mutations exert their effects in different experimental systems, by either increasing Aβ1-42 production, decreasing Aβ1-40 production, or both. Because it could be explained by the heterogeneity in the composition of γ-secretase, we purified to homogeneity complexes made of human NCT, APH1aL, PEN2, and the pathogenic PS1 mutants L166P, ΔE9, or P436Q. METHODOLOGY/PRINCIPAL FINDINGS: We took advantage of a mouse embryonic fibroblast cell line lacking PS1 and PS2 to generate different stable cell lines overexpressing human γ-secretase complexes with different FAD-linked PS1 mutations. A multi-step affinity purification procedure was used to isolate semi-purified or highly purified γ-secretase complexes. The functional characterization of these complexes revealed that all PS1 FAD-linked mutations caused a loss of γ-secretase activity phenotype, in terms of Aβ1-40, Aβ1-42 and APP intracellular domain productions in vitro. CONCLUSION/SIGNIFICANCE: Our data support the view that PS1 mutations lead to a strong γ-secretase loss-of-function phenotype and an increased Aβ1-42/Aβ1-40 ratio, two mechanisms that are potentially involved in the pathogenesis of Alzheimer's disease

Gender- and Age-Dependent γ-Secretase Activity in Mouse Brain and Its Implication in Sporadic Alzheimer Disease

Alzheimer disease (AD) is an age-related disorder. Aging and female gender are two important risk factors associated with sporadic AD. However, the mechanism by which aging and gender contribute to the pathogenesis of sporadic AD is unclear. It is well known that genetic mutations in γ-secretase result in rare forms of early onset AD due to the aberrant production of Aβ42 peptides, which are the major constituents of senile plaques. However, the effect of age and gender on γ-secretase has not been fully investigated. Here, using normal wild-type mice, we show mouse brain γ-secretase exhibits gender- and age-dependent activity. Both male and female mice exhibit increased Aβ42∶Aβ40 ratios in aged brain, which mimics the effect of familial mutations of Presenilin-1, Presenlin-2, and the amyloid precursor protein on Aβ production. Additionally, female mice exhibit much higher γ-secretase activity in aged brain compared to male mice. Furthermore, both male and female mice exhibit a steady decline in Notch1 γ-secretase activity with aging. Using a small molecule affinity probe we demonstrate that male mice have less active γ-secretase complexes than female mice, which may account for the gender-associated differences in activity in aged brain. These findings demonstrate that aging can affect γ-secretase activity and specificity, suggesting a role for γ-secretase in sporadic AD. Furthermore, the increased APP γ-secretase activity seen in aged females may contribute to the increased incidence of sporadic AD in women and the aggressive Aβ plaque pathology seen in female mouse models of AD. In addition, deceased Notch γ-secretase activity may also contribute to neurodegeneration. Therefore, this study implicates altered γ-secretase activity and specificity as a possible mechanism of sporadic AD during aging

Hippocampal Spatial Memory Impairments Caused by the Familial Alzheimer’s Disease-Linked Presenilin 1 M146V Mutation

Mutations in presenilins (PS) 1 and 2 are the major cause of familial Alzheimer's disease. Conditional inactivation of PS1 in the mouse postnatal forebrain leads to mild deficits in spatial learning and memory, whereas inactivation of both PS1 and PS2 results in severe memory and synaptic plasticity impairments, followed by progressive and substantial neurodegeneration. Here we investigate the effect of a familial Alzheimer's disease-linked PS1 missense mutation using knock-in (KI) mice, in which the wild-type PS1 allele is replaced with the M146V mutant allele. In the Morris water maze task, PS1 KI mice at 3 months of age exhibit reduced quadrant occupancy and platform crossing in the probe trial after 6 days of training, though their performance was normal in the probe trial after 12 days of training. By the age of 9 months, even after 12 days of training, PS1 homozygous KI mice still exhibit reduced platform crossing in the post-training probe trial. ELISA analysis revealed a selective increase in cortical levels of beta-amyloid 42 in PS1 KI mice, whereas production of beta-amyloid 40 was normal. Histological and quantitative real-time RT-PCR analyses showed normal gross hippocampal morphology and unaltered expression of three genes involved in inflammatory responses in PS1 KI mice. These results show hippocampal spatial memory impairments caused by the PS1 M146V mutation and age-related deterioration of the memory impairment, suggesting that PS1 KI mice are a valuable model system for the study of memory loss in AD

The DREAMS experiment flown on the ExoMars 2016 mission for the study of Martian environment during the dust storm season

International audienceThe DREAMS (Dust characterization, Risk assessment and Environment Analyser on the Martian Surface) experiment on Schiaparelli lander of ExoMars 2016 mission was an autonomous meteorological station designed to completely characterize the Martian atmosphere on surface, acquiring data not only on temperature, pressure, humidity, wind speed and direction, but also on solar irradiance, dust opacity and atmospheric electrification, to measure for the first time key parameters linked to hazard conditions for future manned explorations. Although with very limited mass and energy resources, DREAMS would be able to operate autonomously for at least two Martian days (sols) after landing in a very harsh environment as it was supposed to land on Mars during the dust storm season (October 2016 in Meridiani Planum) relying on its own power supply. ExoMars mission was successfully launched on 14th March 2016 and Schiaparelli entered the Mars atmosphere on October 20th beginning its ‘six minutes of terror’ journey to the surface. Unfortunately, some unexpected behavior during the parachuted descent caused an unrecoverable critical condition in navigation system of the lander driving to a destructive crash on the surface. The adverse sequence of events at 4 km altitude triggered the transition of the lander in surface operative mode, commanding switch on the DREAMS instrument, which was therefore able to correctly power on and send back housekeeping data. This proved the nominal performance of all DREAMS hardware before touchdown demonstrating the highest TRL of the unit for future missions. This paper describes this experiment in terms of scientific goals, design, performances, testing and operational capabilities with an overview of in flight performances and available mission data

The DREAMS experiment flown on the ExoMars 2016 mission for the study of Martian environment during the dust storm season

The DREAMS (Dust characterization, Risk assessment and Environment Analyser on the Martian Surface) experiment on Schiaparelli lander of ExoMars 2016 mission was an autonomous meteorological station designed to completely characterize the Martian atmosphere on surface, acquiring data not only on temperature, pressure, humidity, wind speed and direction, but also on solar irradiance, dust opacity and atmospheric electrification, to measure for the first time key parameters linked to hazard conditions for future manned explorations. Although with very limited mass and energy resources, DREAMS would be able to operate autonomously for at least two Martian days (sols) after landing in a very harsh environment as it was supposed to land on Mars during the dust storm season (October 2016 in Meridiani Planum) relying on its own power supply. ExoMars mission was successfully launched on 14th March 2016 and Schiaparelli entered the Mars atmosphere on October 20th beginning its `six minutes of terror' journey to the surface. Unfortunately, some unexpected behavior during the parachuted descent caused an unrecoverable critical condition in navigation system of the lander driving to a destructive crash on the surface. The adverse sequence of events at 4 km altitude triggered the transition of the lander in surface operative mode, commanding switch on the DREAMS instrument, which was therefore able to correctly power on and send back housekeeping data. This proved the nominal performance of all DREAMS hardware before touchdown demonstrating the highest TRL of the unit for future missions. This paper describes this experiment in terms of scientific goals, design, performances, testing and operational capabilities with an overview of in flight performances and available mission data