- Email: [email protected]
P1-094: Mouse resources for the study of neurodegenerative diseases
T234
Poster Presentations P1
ternation in Y-maze. Importantly, BACE1⫹/- mutation ameliorated the synaptic and cognitive impairments found in APP/PS1 transgenic mice at 6 months of age. We are currently conducting biochemical experiments, and potential correlations between the functional improvements and reductions of cerebral Abeta species in BACE1⫹/-;APP/PS1 mice will also be discussed. Conclusions: Our findings support the notion that partial inhibition of BACE1 may be sufficient to rescue AD-related synaptic and memory deficits. P1-093
GENDER-SPECIFIC DEVELOPMENT OF BRAIN PATHOLOGY AND FUNCTIONAL DEFICITS IN A hAPPSL MOUSE MODEL OF ALZHEIMER’S DISEASE (TASD41)
Michael Hierzer, Manuela Prokesch, Maria Posch, Stephan Duller, Birgit Hutter-Paier, Manfred Windisch, JSW CNS Research, Graz, Austria. Contact e-mail: [email protected] Background: Alzheimer’s disease (AD) results in massive neuronal degeneration, causing memory loss, disorientation, loss of language skills, learning difficulties, and decline in the ability to meet requirements of daily living. AD is pathologically characterized by amyloid plaques and neurofibrillary tangles. In spite of intense research efforts there is still a lack of knowledge about the etiology and pathogenesis. Animal models provide a useful tool for understanding the complexity of neurodegenerative processes and for the development of treatment strategies proposed to counteract the disease. Methods: We have established a transgenic mouse model over-expressing human (h) APP751 containing the London (V717I) and Swedish (K670M/N671L) mutations under the regulatory control of the murine Thy-1 promoter (mThy-1-hAPP751). Behavior of this APP751SL mouse model was extensively studied using different age groups of male and female mice. Brain pathological changes were investigated immunohistochemically and biochemically. Results: Compared to non-transgenic littermates behavioral deficits occurred in terms of learning and memory in the Hole Board Test, the Morris Water Maze, the Object Recognition and the Contextual Fear Conditioning task, showing an age and to a certain degree also a gender related progression. Brain amyloid plaque load, as well as soluble and bound A1-40 and A1-42 brain levels significantly increased over age in these mice. Brain histopathology started with formation of amyloid plaques in the cortex at 3 to 4 months of age, reaching the hippocampus at 6 to 7 months. Females being earlier and more affected than males. Like in humans plaque load is accompanied by brain inflammatory processes, synaptic alterations and neuronal changes. Conclusions: Due to the progressive behavioral and neuropathological changes this APP751SL mouse model presents a valid model system for AD. P1-094
MOUSE RESOURCES FOR THE STUDY OF NEURODEGENERATIVE DISEASES
Michael Sasner, Steve Rockwood, Cathy Lutz, Leah Rae Donahue, The Jackson Lab, Bar Harbor, ME, USA. Contact e-mail: [email protected] Background: Mouse models are critical tools to study the genetics and pathogenesis of Alzheimer’s Disease (AD) and can be used to develop new therapies. The Alzheimer’s Disease Mouse Model Resource (ADMMR) was established at The Jackson Laboratory Repository to be a central resource for archiving and distributing AD models at a high health status. The Repository also backcrosses strains to standard genetic backgrounds, and provides information and database search tools to aid in the selection and use of mouse models. Methods: Submission of a strain to the Repository fulfills requirements for sharing of mice in accordance with NIH’s policy for sharing of research reagents; strains can be submitted using an online web form. If necessary, donating investigators can place use restrictions on their strains. Results: ADMMR holdings continue to grow, and now include more than 50 models carrying mutations in 13 different genes directly relevant to AD, including apolipoprotein E (Apoe), amyloid beta precursor protein (App), beta-site APP cleaving enzymes (Bace1 and
Bace2), a regulatory subunit of cyclin-dependent kinase 5 (Cdk5r1), microtubule-associated protein tau (Mapt), and presenilins (Psen1 and Psen2). Some of these transgenes are expressed in an inducible manner. In most cases studied to date, mice carrying combinations of mutant alleles display a more severe disease phenotype; the ADMMR maintains some of these multi-allele strains, and enables researchers to create new combinations of alleles. The more widely used disease models are available as aged (at least six months old) mice. Database search tools allow strains to be identified by mutation, phenotype, or relationship to human disease. The Repository also distributes transgenic mice that express cre or reporter genes in a neuronal-specific manner that can be used in neurodegeneration research. These lines are now searchable by expression site to make it easier to find the appropriate line for a specific purpose. In addition, the Repository distributes mouse models relevant to the study of Parkinson’s and Huntington’s Diseases. Conclusions: For more information about the ADMMR including a list of strains, allele and phenotypic information, and associated references, see http://jaxmice.jax.org/research/alzheimers.html. The ADMMR is supported by the NCRR and an anonymous foundation. P1-095
CNS STEM CELL LINES FROM TAU TRANSGENIC MICE AS MODELS FOR FRONTOTEMPORAL DEMENTIA
Miranda E. Orr1,2, Ranjit K. Giri1, Karen H. Ashe3, George A. Carlson1, 1McLaughlin Research Institute, Great Falls, MT, USA; 2Montana State University, Bozeman, MT, USA; 3N. Bud Grossman Center for Memory Research and Care, University of Minnesota, Minneapolis, MN, USA. Contact e-mail: [email protected] Background: Human stem cell lines that capture the genetics of disease susceptibility provide new research tools. To assess the utility of stem cells to study neurodegenerative disease we are using mouse central nervous system stem cell-containing neurosphere cultures. We previously have used this system to study aspects of prion disease in vitro and are now investigating the utility of neurospheres to model tauopathies, which, like prion diseases, arise from protein misprocessing events. Tauopathies are characterized by hyperphosphorylation, aggregation, and neurofibrillary tangle formation of the microtubule associated protein tau. Methods: Transgenic mice expressing a mutant human tauP301L transgene under the control of a tetracycline responsive element are crossed to transgenic mice that express the tetracycline transactivator under control of the calcium calmodulin kinase promoter. The resulting rTg(tauP301L)4510 (rTg4510) mice display cellular, histological, biochemical and behavioral phenotypes similar to frontotemporal dementia in humans. Neurosphere cultures derived from rTg4510 mice will be compared to cultures derived from similar mice that express the human wild type tau transgene, rTg(tauwt)21221 (rTg21221). Results: We have generated neurosphere lines from embryonic day 14 rTg4510, rTg21221, and non-expressing mice. Human tau expression is detected by Western blot and immunofluorescence in tauP301L and tauwt cell cultures but not in controls. Human tau expression is maintained over several passages in both undifferentiated neurospheres and differentiated cells derived from them. Human tau expression does not alter cell growth rates regardless of human tau genotype. However, tauP301L transgene expression may affect cell types in the culture following differentiation stimuli. Using immunofluorescence, both glial and neuronal markers are observed post-differentiation in all genotypes. The proportion of cells co-expressing neuron-specific antigens and human tau is higher in tauwt expressing cultures than in tauP301L expressing cultures. The number of cells co-expressing glial antigens and human tau transgene, however, are similar among culture genotypes. We are now investigating tau pathology in cultured neurospheres using tau hyperphosphorylation and aggregation as surrogates for disease progression in vitro. Conclusions: Our results suggest that tauP301L expression affects neurosphere cultures differently from tauwt expression and that stem cell cultures may be useful models for neurodegenerative diseases.
Poster Presentations P1
ternation in Y-maze. Importantly, BACE1⫹/- mutation ameliorated the synaptic and cognitive impairments found in APP/PS1 transgenic mice at 6 months of age. We are currently conducting biochemical experiments, and potential correlations between the functional improvements and reductions of cerebral Abeta species in BACE1⫹/-;APP/PS1 mice will also be discussed. Conclusions: Our findings support the notion that partial inhibition of BACE1 may be sufficient to rescue AD-related synaptic and memory deficits. P1-093
GENDER-SPECIFIC DEVELOPMENT OF BRAIN PATHOLOGY AND FUNCTIONAL DEFICITS IN A hAPPSL MOUSE MODEL OF ALZHEIMER’S DISEASE (TASD41)
Michael Hierzer, Manuela Prokesch, Maria Posch, Stephan Duller, Birgit Hutter-Paier, Manfred Windisch, JSW CNS Research, Graz, Austria. Contact e-mail: [email protected] Background: Alzheimer’s disease (AD) results in massive neuronal degeneration, causing memory loss, disorientation, loss of language skills, learning difficulties, and decline in the ability to meet requirements of daily living. AD is pathologically characterized by amyloid plaques and neurofibrillary tangles. In spite of intense research efforts there is still a lack of knowledge about the etiology and pathogenesis. Animal models provide a useful tool for understanding the complexity of neurodegenerative processes and for the development of treatment strategies proposed to counteract the disease. Methods: We have established a transgenic mouse model over-expressing human (h) APP751 containing the London (V717I) and Swedish (K670M/N671L) mutations under the regulatory control of the murine Thy-1 promoter (mThy-1-hAPP751). Behavior of this APP751SL mouse model was extensively studied using different age groups of male and female mice. Brain pathological changes were investigated immunohistochemically and biochemically. Results: Compared to non-transgenic littermates behavioral deficits occurred in terms of learning and memory in the Hole Board Test, the Morris Water Maze, the Object Recognition and the Contextual Fear Conditioning task, showing an age and to a certain degree also a gender related progression. Brain amyloid plaque load, as well as soluble and bound A1-40 and A1-42 brain levels significantly increased over age in these mice. Brain histopathology started with formation of amyloid plaques in the cortex at 3 to 4 months of age, reaching the hippocampus at 6 to 7 months. Females being earlier and more affected than males. Like in humans plaque load is accompanied by brain inflammatory processes, synaptic alterations and neuronal changes. Conclusions: Due to the progressive behavioral and neuropathological changes this APP751SL mouse model presents a valid model system for AD. P1-094
MOUSE RESOURCES FOR THE STUDY OF NEURODEGENERATIVE DISEASES
Michael Sasner, Steve Rockwood, Cathy Lutz, Leah Rae Donahue, The Jackson Lab, Bar Harbor, ME, USA. Contact e-mail: [email protected] Background: Mouse models are critical tools to study the genetics and pathogenesis of Alzheimer’s Disease (AD) and can be used to develop new therapies. The Alzheimer’s Disease Mouse Model Resource (ADMMR) was established at The Jackson Laboratory Repository to be a central resource for archiving and distributing AD models at a high health status. The Repository also backcrosses strains to standard genetic backgrounds, and provides information and database search tools to aid in the selection and use of mouse models. Methods: Submission of a strain to the Repository fulfills requirements for sharing of mice in accordance with NIH’s policy for sharing of research reagents; strains can be submitted using an online web form. If necessary, donating investigators can place use restrictions on their strains. Results: ADMMR holdings continue to grow, and now include more than 50 models carrying mutations in 13 different genes directly relevant to AD, including apolipoprotein E (Apoe), amyloid beta precursor protein (App), beta-site APP cleaving enzymes (Bace1 and
Bace2), a regulatory subunit of cyclin-dependent kinase 5 (Cdk5r1), microtubule-associated protein tau (Mapt), and presenilins (Psen1 and Psen2). Some of these transgenes are expressed in an inducible manner. In most cases studied to date, mice carrying combinations of mutant alleles display a more severe disease phenotype; the ADMMR maintains some of these multi-allele strains, and enables researchers to create new combinations of alleles. The more widely used disease models are available as aged (at least six months old) mice. Database search tools allow strains to be identified by mutation, phenotype, or relationship to human disease. The Repository also distributes transgenic mice that express cre or reporter genes in a neuronal-specific manner that can be used in neurodegeneration research. These lines are now searchable by expression site to make it easier to find the appropriate line for a specific purpose. In addition, the Repository distributes mouse models relevant to the study of Parkinson’s and Huntington’s Diseases. Conclusions: For more information about the ADMMR including a list of strains, allele and phenotypic information, and associated references, see http://jaxmice.jax.org/research/alzheimers.html. The ADMMR is supported by the NCRR and an anonymous foundation. P1-095
CNS STEM CELL LINES FROM TAU TRANSGENIC MICE AS MODELS FOR FRONTOTEMPORAL DEMENTIA
Miranda E. Orr1,2, Ranjit K. Giri1, Karen H. Ashe3, George A. Carlson1, 1McLaughlin Research Institute, Great Falls, MT, USA; 2Montana State University, Bozeman, MT, USA; 3N. Bud Grossman Center for Memory Research and Care, University of Minnesota, Minneapolis, MN, USA. Contact e-mail: [email protected] Background: Human stem cell lines that capture the genetics of disease susceptibility provide new research tools. To assess the utility of stem cells to study neurodegenerative disease we are using mouse central nervous system stem cell-containing neurosphere cultures. We previously have used this system to study aspects of prion disease in vitro and are now investigating the utility of neurospheres to model tauopathies, which, like prion diseases, arise from protein misprocessing events. Tauopathies are characterized by hyperphosphorylation, aggregation, and neurofibrillary tangle formation of the microtubule associated protein tau. Methods: Transgenic mice expressing a mutant human tauP301L transgene under the control of a tetracycline responsive element are crossed to transgenic mice that express the tetracycline transactivator under control of the calcium calmodulin kinase promoter. The resulting rTg(tauP301L)4510 (rTg4510) mice display cellular, histological, biochemical and behavioral phenotypes similar to frontotemporal dementia in humans. Neurosphere cultures derived from rTg4510 mice will be compared to cultures derived from similar mice that express the human wild type tau transgene, rTg(tauwt)21221 (rTg21221). Results: We have generated neurosphere lines from embryonic day 14 rTg4510, rTg21221, and non-expressing mice. Human tau expression is detected by Western blot and immunofluorescence in tauP301L and tauwt cell cultures but not in controls. Human tau expression is maintained over several passages in both undifferentiated neurospheres and differentiated cells derived from them. Human tau expression does not alter cell growth rates regardless of human tau genotype. However, tauP301L transgene expression may affect cell types in the culture following differentiation stimuli. Using immunofluorescence, both glial and neuronal markers are observed post-differentiation in all genotypes. The proportion of cells co-expressing neuron-specific antigens and human tau is higher in tauwt expressing cultures than in tauP301L expressing cultures. The number of cells co-expressing glial antigens and human tau transgene, however, are similar among culture genotypes. We are now investigating tau pathology in cultured neurospheres using tau hyperphosphorylation and aggregation as surrogates for disease progression in vitro. Conclusions: Our results suggest that tauP301L expression affects neurosphere cultures differently from tauwt expression and that stem cell cultures may be useful models for neurodegenerative diseases.