- Email: [email protected]
P2-027 Effect and mechanisms of constant light-illumination on tau phosphorylation in rat hippocampus and the protection of melatonin
Poster Session P2: Animal and Cellular Models - Animal Models, Non-transgenic their knowledge of the exact platform location was less precise than young rats. Five day later, aged rats demonstrated impaired memory retention as indicated by the memory index and again their proximity score indicated less accurate knowledge of the platform's location. Conclusions: We have designed and validated a new set of sensitive and novel memory measures for use in the RAWM. These methods will greatly aid the behavioural analysis of aged and transgenic animals used in studies of Alzheimer's disease.
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D E F I C I T O F SPATIAL L E A R N I N G A N D M E M O R Y OF THE YOUNG ACCELERATED SENESCENCE P R O N E M O U S E 8 IN T H E R A D I A L S I X - A R M W A T E R
MAZE
These rates reflect the half-lives of proteins being studied, or the growth and death rate of cells in isolated tissues. Results: Techniques have been developed for the analysis of nanogram amounts of alanine by GC/MS, as well as sub-nanogram amounts of deoxyribose. We have isolated sufficient quantities of APP for GC/MS analysis from mouse brains by ion exchange, heparin, and hydrophobic interaction chromatography. We have also detected sufficient CTF (C99) in mouse brains for analysis. Ample DNA for GC/MS analysis has been isolated from 20 mg of hippocampal tissue. Kinetic results (synthesis and turnover rates) of these components in brain will be presented. Conclusions: Kinetic studies of the amyloid beta pathway and neural cell growth in normal and transgenic mice are now possible, and may provide insight into fundamental processes of Alzheimer's disease.
Gui H. Chen* 1, Yue J. Wang 1, Jiang N. Zhou 2, Xiao M. Wang 3. 1Anhui
•P2f•
Geriatric Institute, the First Affiliated Hospital of Anhui Medical University, Hefei, China; 2 University of Science and Technology of China, Hefei, China; 3Institute of Neuroscience, Beijing University, Beijing, China. Contact e-mail: [email protected]
Ling Z. Qun*. Tongji Medical College, Wuhan, Hubei, China. Contact
Background: Available data show that impairment in the non-spatial learning and memory of the senescence-accelerated prone mouse 8 (SAMP8), an appropriate model of brain aging, begins as early as 2 months old; however the deficiency of the spatial learning and memory begins as late as four months old above. However, this is inconsistent with the results from other strains (e.g. C57BL mice). Measure apparatuses might be responsible for the discrepancy. Objective: To determine whether the impairment of the spatial learning and memory exists in young SAMP8. Methods: Radial six-arm water maze (RAWM), a new testing apparatus for detecting the spatial learning and memory in mice, and Morris maze, a classic spatial learning and memory testing apparatus. Results: It was found that, just in the RAWM rather than in the Morris maze, the impairment of the spatial learning and/or the memory could be measured in SAMP8 mice as early as three months old. The measures stemmed from the RAWM and the Morris maze respectively were associated with different factors through the principle components analysis, Conclusions: The defect of spatial learning and memory could be found in early life of SAMP8 mice, and RAWM and Morris maze could detect the different aspects of spatial learning and memory.
P2•P•-026•
Michael E. Marino*, Mahalakshmi Shankaran, Carole Black, Mohamad Awada, Ablatt Mahsut, Marc Hellerstein. Kinemed, Inc.,
Emeryville, CA, USA. Contact e-mail: [email protected]
Background: Stable isotope incorporation allows for the kinetic study of targeted cells and proteins. Kinetic information (half-life, production rates, tissue residence times) from disease-related molecular targets often gives insight into mechanisms of disease when static measurements cannot. Kinetic studies of the Amyloid precursor protein (APP), APP C-terminal fragment (CTF, C99), and neural progenitor cells in mouse models may give insight into the mechanisms of AD. Objective(s): The objective of this work is to determine the turnover rate of APP, CTF (C99), and neural progenitor cells in normal and transgenic (APP-Swe - Tg2576) mice. The intent is to discover the kinetic determinants of AD in mouse models, and to establish useful biomarkers for the study of AD therapeutics. Methods: Tg2576 and normal mice are given a bolus of deuterated water to achieve 5% 2H20 enrichment, and then drink 8% deuterated water for a four week labeling period. Deuterium from 2H20 is incorporated into the deoxyribose of newly synthesized DNA and into non-essential amino acids of protein during labeling. APP, CTF, and hippocampal DNA are isolated from mice sacrificed throughout the study. Isolated material is hydrolyzed and derivatized, and deuterium incorporation into deoxyribose or alanine is determined by GC/MS. From this, fractional synthesis rates of APP and CTE and cell proliferation in the hippocampus are determined.
EFFECT AND MECHANISMS OF CONSTANT L I G H T - I L L U M I N A T I O N ON TAU P H O S P H O R Y L A T I O N IN RAT H I P P O C A M P U S A N D THE PROTECTION OF MELATONIN
e-mail: [email protected]
Objective: To study whether the melatonin (MEL) depletion induces Alzheimer-like tan hyperphosphorylation and explores the possible underlying mechanisms. Methods: Constantly light-illuminated rats with or without exogenous supplementation of MEL for one month. Results: Protein tan was abnormally hyperphosphorylated at Ser396/Ser404 (PHF-1) and Ser214 (P214) in hippocampus extracts of the light-induced rats, and a significantly diminished MEL and a remarkably elevated dopamine as well as a significantly activated protein kinase A (PKA) and inhibited protein phosphatase-1 (PP-I) were observed in these model rats. Daily supplementation of l mmol/L but not 10 mmol/L of MEL partially restores serum level of MEL and dopamine, suppresses PKA overactivation and relieves PP-1 inhibition, and thus blocks tan hyperphosphorylation at PHF-1 and P214 epitopes. Conclusion: Constant light-illumination induces tan hyperphosphorylation in vivo via most dominantly overactivation of PKA and inhibition of PP-1, which might be resulted from MEL deficit and/or dopamine accumulation. Key words: Alzheimer disease, light-illumination, melatonin, tan, hyperphosphorylation
• KINETIC STUDIES OF THE AMYLOID-BETA PATHWAY AND C O N C U R R E N T M E A S U R E M E N T OF N E U R O G E N E S I S F O R T H E S T U D Y OF AD
$231
BOTH, E X E R C I S E A N D E N R I C H E D E N V I R O N M E N T ENHANCE THE COGNITIVE IMPROVEMENT RELATED TO STEM CELLS GRAFTING TO THE I M P A I R E D A G E D BRAIN: AN APPROACH T O M U L T I F A C T O R I A L T H E R A P Y IN A L Z H E I M E R DISEASE
Caridad I. Fernandez* 1, Esteban Alberti 2, Jaime Collazo 1, Maria E. Gonzalez 1, Jose Y. Bauza 1, Lisis Martinez 1, Juan C. Rosillo 1. lIntl Ctr
Neurological, Havana City, Cuba; 2Intl Ctr Neurological Neurol Restoration (CIREN), Havana City, Cuba. Contact e-mail: ivettefer@yahoo, es
Background: The selective loss and/or dystrophy of specific subsets of neuronal populations are key features in Alzheimer's disease (AD). Research showing that (i) neurogenesis normally occurs in limited regions of the adult mammalian brain (ii) that there are significant numbers of multipotent neural precursors in many parts of the adult brain (iii) that it is possible to induce functional recovery even in the aged brain, via manipulation of endogenous precursors in situ with stimulus i.e. enriched environment (EE) and exercise (Ex) and; (iv) that nonneural stem cells can survive and differentiate to neurons after brain grafting. Objective: We examined the effects of mesenchymai stem cells (MSCs) -labelled with bis-benzimide- grafted to the cortex or hippocampus of cognitively impaired aged rats simultaneously submitted to EE or Ex. Methods: Additional groups received EE or Ex with negative grafting (no MSCs). Behavioural analysis includes Morris Water Maze, avoidance test and locomotor/sensorimotor tests. Eight weeks after it, all rats were re-evaluated at behavioural tests mad sacrificed. Half/half of brains were processed for immunostaining mad fluorescence microscopy
•
D E F I C I T O F SPATIAL L E A R N I N G A N D M E M O R Y OF THE YOUNG ACCELERATED SENESCENCE P R O N E M O U S E 8 IN T H E R A D I A L S I X - A R M W A T E R
MAZE
These rates reflect the half-lives of proteins being studied, or the growth and death rate of cells in isolated tissues. Results: Techniques have been developed for the analysis of nanogram amounts of alanine by GC/MS, as well as sub-nanogram amounts of deoxyribose. We have isolated sufficient quantities of APP for GC/MS analysis from mouse brains by ion exchange, heparin, and hydrophobic interaction chromatography. We have also detected sufficient CTF (C99) in mouse brains for analysis. Ample DNA for GC/MS analysis has been isolated from 20 mg of hippocampal tissue. Kinetic results (synthesis and turnover rates) of these components in brain will be presented. Conclusions: Kinetic studies of the amyloid beta pathway and neural cell growth in normal and transgenic mice are now possible, and may provide insight into fundamental processes of Alzheimer's disease.
Gui H. Chen* 1, Yue J. Wang 1, Jiang N. Zhou 2, Xiao M. Wang 3. 1Anhui
•P2f•
Geriatric Institute, the First Affiliated Hospital of Anhui Medical University, Hefei, China; 2 University of Science and Technology of China, Hefei, China; 3Institute of Neuroscience, Beijing University, Beijing, China. Contact e-mail: [email protected]
Ling Z. Qun*. Tongji Medical College, Wuhan, Hubei, China. Contact
Background: Available data show that impairment in the non-spatial learning and memory of the senescence-accelerated prone mouse 8 (SAMP8), an appropriate model of brain aging, begins as early as 2 months old; however the deficiency of the spatial learning and memory begins as late as four months old above. However, this is inconsistent with the results from other strains (e.g. C57BL mice). Measure apparatuses might be responsible for the discrepancy. Objective: To determine whether the impairment of the spatial learning and memory exists in young SAMP8. Methods: Radial six-arm water maze (RAWM), a new testing apparatus for detecting the spatial learning and memory in mice, and Morris maze, a classic spatial learning and memory testing apparatus. Results: It was found that, just in the RAWM rather than in the Morris maze, the impairment of the spatial learning and/or the memory could be measured in SAMP8 mice as early as three months old. The measures stemmed from the RAWM and the Morris maze respectively were associated with different factors through the principle components analysis, Conclusions: The defect of spatial learning and memory could be found in early life of SAMP8 mice, and RAWM and Morris maze could detect the different aspects of spatial learning and memory.
P2•P•-026•
Michael E. Marino*, Mahalakshmi Shankaran, Carole Black, Mohamad Awada, Ablatt Mahsut, Marc Hellerstein. Kinemed, Inc.,
Emeryville, CA, USA. Contact e-mail: [email protected]
Background: Stable isotope incorporation allows for the kinetic study of targeted cells and proteins. Kinetic information (half-life, production rates, tissue residence times) from disease-related molecular targets often gives insight into mechanisms of disease when static measurements cannot. Kinetic studies of the Amyloid precursor protein (APP), APP C-terminal fragment (CTF, C99), and neural progenitor cells in mouse models may give insight into the mechanisms of AD. Objective(s): The objective of this work is to determine the turnover rate of APP, CTF (C99), and neural progenitor cells in normal and transgenic (APP-Swe - Tg2576) mice. The intent is to discover the kinetic determinants of AD in mouse models, and to establish useful biomarkers for the study of AD therapeutics. Methods: Tg2576 and normal mice are given a bolus of deuterated water to achieve 5% 2H20 enrichment, and then drink 8% deuterated water for a four week labeling period. Deuterium from 2H20 is incorporated into the deoxyribose of newly synthesized DNA and into non-essential amino acids of protein during labeling. APP, CTF, and hippocampal DNA are isolated from mice sacrificed throughout the study. Isolated material is hydrolyzed and derivatized, and deuterium incorporation into deoxyribose or alanine is determined by GC/MS. From this, fractional synthesis rates of APP and CTE and cell proliferation in the hippocampus are determined.
EFFECT AND MECHANISMS OF CONSTANT L I G H T - I L L U M I N A T I O N ON TAU P H O S P H O R Y L A T I O N IN RAT H I P P O C A M P U S A N D THE PROTECTION OF MELATONIN
e-mail: [email protected]
Objective: To study whether the melatonin (MEL) depletion induces Alzheimer-like tan hyperphosphorylation and explores the possible underlying mechanisms. Methods: Constantly light-illuminated rats with or without exogenous supplementation of MEL for one month. Results: Protein tan was abnormally hyperphosphorylated at Ser396/Ser404 (PHF-1) and Ser214 (P214) in hippocampus extracts of the light-induced rats, and a significantly diminished MEL and a remarkably elevated dopamine as well as a significantly activated protein kinase A (PKA) and inhibited protein phosphatase-1 (PP-I) were observed in these model rats. Daily supplementation of l mmol/L but not 10 mmol/L of MEL partially restores serum level of MEL and dopamine, suppresses PKA overactivation and relieves PP-1 inhibition, and thus blocks tan hyperphosphorylation at PHF-1 and P214 epitopes. Conclusion: Constant light-illumination induces tan hyperphosphorylation in vivo via most dominantly overactivation of PKA and inhibition of PP-1, which might be resulted from MEL deficit and/or dopamine accumulation. Key words: Alzheimer disease, light-illumination, melatonin, tan, hyperphosphorylation
• KINETIC STUDIES OF THE AMYLOID-BETA PATHWAY AND C O N C U R R E N T M E A S U R E M E N T OF N E U R O G E N E S I S F O R T H E S T U D Y OF AD
$231
BOTH, E X E R C I S E A N D E N R I C H E D E N V I R O N M E N T ENHANCE THE COGNITIVE IMPROVEMENT RELATED TO STEM CELLS GRAFTING TO THE I M P A I R E D A G E D BRAIN: AN APPROACH T O M U L T I F A C T O R I A L T H E R A P Y IN A L Z H E I M E R DISEASE
Caridad I. Fernandez* 1, Esteban Alberti 2, Jaime Collazo 1, Maria E. Gonzalez 1, Jose Y. Bauza 1, Lisis Martinez 1, Juan C. Rosillo 1. lIntl Ctr
Neurological, Havana City, Cuba; 2Intl Ctr Neurological Neurol Restoration (CIREN), Havana City, Cuba. Contact e-mail: ivettefer@yahoo, es
Background: The selective loss and/or dystrophy of specific subsets of neuronal populations are key features in Alzheimer's disease (AD). Research showing that (i) neurogenesis normally occurs in limited regions of the adult mammalian brain (ii) that there are significant numbers of multipotent neural precursors in many parts of the adult brain (iii) that it is possible to induce functional recovery even in the aged brain, via manipulation of endogenous precursors in situ with stimulus i.e. enriched environment (EE) and exercise (Ex) and; (iv) that nonneural stem cells can survive and differentiate to neurons after brain grafting. Objective: We examined the effects of mesenchymai stem cells (MSCs) -labelled with bis-benzimide- grafted to the cortex or hippocampus of cognitively impaired aged rats simultaneously submitted to EE or Ex. Methods: Additional groups received EE or Ex with negative grafting (no MSCs). Behavioural analysis includes Morris Water Maze, avoidance test and locomotor/sensorimotor tests. Eight weeks after it, all rats were re-evaluated at behavioural tests mad sacrificed. Half/half of brains were processed for immunostaining mad fluorescence microscopy