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Tauopathy in zebrafish using life imaging transgenic zebrafish
Symposia S1-03: Prevention of Dementia S1-02-04
TAUOPATHY IN ZEBRAFISH USING LIFE IMAGING TRANSGENIC ZEBRAFISH
Dominik Paquet1, Gabriela Plucinska2, Leanne Godinho2, Bettina Schmid1, Christian Haass1, Thomas Misgeld2, 1DZNE Munich, Munich, Germany; 2TU Munich, Munich, Germany. Background: Axonal transport is a pivotal process for neurons. Transport defects can lead to a deficiency of essential proteins and accumulation of cellular debris at areas distant from the soma, leading to starvation and malfunction of synapses and finally neurodegeneration. Axonal transport deficits have been described in neurodegenerative diseases, including AD and FTD. The Tau protein, which plays a central role in both diseases as hyperphosphorylated constituent of neurofibrillary tangles, is important for regulating microtubules, the intracellular transport tracks. Transport alterations appear to occur early in the cascade of neurological damage, long before neurons die. Therefore, axonal transport represents a promising drug target for neurodegenerative diseases. Mechanisms of axonal transport have been mainly studied in cultured neurons. While this approach has greatly expanded our knowledge of transport phenomena, it is also limited, as in vitro systems neither fully reflect the geometrical complexity of neurons in vivo and their interplay with other cells, nor easily mimic processes related to human disease. More intact preparations derived from invertebrates or mice circumvented some of these problems, but more versatile systems that allow studying axonal transport in vivo, together with screening for modulators, are still needed. Methods: We addressed this by developing an in vivo ‘tool box’ to study axonal transport in optically transparent and genetically accessible zebrafish. We have established the characteristics of mitochondrial transport in individually labelled neurons. Furthermore, we generated transgenic ‘mito-fish’ with fluorescently labelled neuronal mitochondria and performed quantitative measurements of mitochondrial density and transport in normal and compoundtreated animals. Results: As ‘proof-of-principle’, we crossed the ‘mitofish’ to a previously developed zebrafish tauopathy-model, which recapitulates key pathological features of FTD, including hyperphosphorylation, tangle formation and cell death (Paquet et al. 2009). Tau-transgenic fish show a prominent reduction in mitochondrial density and movements in peripheral neurons, which can be rescued by over-expression of MARK, a kinase that regulates Tau binding to microtubules. Conclusions: These approaches allow imaging transport of mitochondria in intact animals with high efficiency, and relating transport to the physiology of complete neurons in their natural habitat. Furthermore, ‘mito-fish’ are useful tools to assess mechanisms of transport disruptions and screen for modulating compounds.
S87
^ Abeta1-40/42 + CFA/IFA (n KLH+ CFA/IFA (n ¼ 8 Tg, 6 WT), 100 Amg ¼ 5 Tg, 8 WT), or vehicle (CFA/IFA; n ¼ 5 Tg, 6 WT) on days 0, 14, and 28 and monthly thereafter. Plasma was collected periodically. Splenocytes were cultured and restimulated with various Abeta peptides. Brains were examined pathologically and by ELISA. Results: J20tg and WT mice generated high antibody levels against the immunizing peptides. Antibodies from mice immunized with Abeta pE3-9 recognized Abeta pE340, 3-9, and 3-13 peptides; splenocytes proliferated upon restimulation with Abeta pE3-9/KLH, KLH, pE3-40and pE3-9 but not Abeta1-40/42 peptides. Antibodies from mice immunized with Abeta1-40/42 recognized Abeta1-40, 1-7 and 1-15 peptides; splenocytes proliferated upon restimulation with Abeta1-40/42 but not the Abeta pE3 peptides or KLH. IgG1 was the dominant isotype for both vaccines. Hippocampal fibrillar plaques were strongly reduced by immunization with Abeta pE3-9 (65%, p ¼ 0.05) and Abeta1-40/42 (69%, p ¼ 0.06) in the J20 tg mice compared to vehicle controls. AbetaX-42 levels were reduced in guanidine HCL brain homogenates of mice immunized by Abeta pE3-9 (28%, p ¼ 0.06) and Abeta1-40/42 (74%, p ¼ 0.002). Abeta pE3 levels were reduced by both vaccines but only reached significance in Abeta1-40/42 immunized mice. Both vaccines significantly increased plasma AbetaX-40 and lowered gliosis. Microhemorrhage was not observed, possibly due to low vascular amyloid at 14 months of age Conclusions: PyroGlu-3 Abeta is a strong immunogen that generates antibodies that lower plaque burden in APP tg mouse brain. Selective removal of toxic Abeta pE3-X may be beneficial for AD.
S1-02-06
TRANSGENIC MODELS TO ADDRESS TOXICITY
Kun Ping Lu, Harvard Medical School, Boston, Mass., United States. Abstract: not available.
SUNDAY, JULY 17, 2011 SYMPOSIA S1-03 PREVENTION OF DEMENTIA S1-03-01
EXERCISE, COGNITION AND THE BRAIN
Arthur Kramer, University of Illinois at Urbana-Champaign, Urbana-Champaign, Ill., United States. S1-02-05
ACTIVE IMMUNIZATION AGAINST PYROGLU-3 Ab GENERATES HIGH ANTIBODY TITERS AND LOWERS PLAQUE BURDEN IN APP TRANSGENIC MICE
Cynthia A Lemere1, Jeffrey Frost2, Martin Kleinschmidt3, Bin Liu2, Qiaoqiao Shi2, Hans-Ulrich Demuth4, Stephan Schilling5, 1Brigham & Women’s Hospital; Harvard Medical School, Boston, Massachusetts; 2 Brigham & Women’s Hospital, Boston, Massachusetts; 3Probiodrug AG, Halle (Saale); 4Probiodrug AG, Halle (Saale); 5Probiodrug AG, Halle. Background: Pyroglutamate-3 Abeta (Abeta pE3-X) is an N-terminally modified Abeta species that is abundant in human plaques and vascular amyloid. Abeta pE3 aggregates quickly, is resistant to degradation, and is neurotoxic. Abeta pE3 is present in AD-like transgenic (tg) mouse models, albeit at lower levels than Abeta1-X. In this pilot study, we compared the immune response and pathological efficacy of active immunization with Abeta pE3-9 vs. Abeta1-40/42 in APP tg and WT mice. Methods: J20 APP tg mice and WT mice of the same strain (B6D2F1) were actively immunized from 6 ^ Abeta pE3-9/ to 14 mo of age. Mice were given i.p. injections of 100 Amg
Background: Populations throughout the industrialized world are becoming increasing sedentary as a result of the changing nature of work and leisure activities. As a result of these societal changes increases in diseases such as hypertension, diabetes, osteoporosis, and forms of cancer are increasing. Physical activity serves to reduce susceptibility to these diseases. However, increased physical activity also has direct and relatively rapid effects on cognition and brain health. Such results have now been reported, over the course of several decades, in animal studies of physical activity. In my presentation I will review research conducted in our laboratory, and the field in general, which has examined the extent to which fitness training enhances cognition and brain structure and function of older adults. The presentation will cover both cross-sectional and intervention studies of fitness differences and fitness training. Studies which assess cognition via both behavioral measures and non-invasive neuroimaging measures, such as magnetic resonance imaging, functional magnetic resonance imaging, eventrelated brain potentials, and the event-related optical signal, will be reviewed and discussed. Finally, I will explore the gaps in the human and animal literature on cognitive and brain health and the manner in which they can be addressed in future research.
TAUOPATHY IN ZEBRAFISH USING LIFE IMAGING TRANSGENIC ZEBRAFISH
Dominik Paquet1, Gabriela Plucinska2, Leanne Godinho2, Bettina Schmid1, Christian Haass1, Thomas Misgeld2, 1DZNE Munich, Munich, Germany; 2TU Munich, Munich, Germany. Background: Axonal transport is a pivotal process for neurons. Transport defects can lead to a deficiency of essential proteins and accumulation of cellular debris at areas distant from the soma, leading to starvation and malfunction of synapses and finally neurodegeneration. Axonal transport deficits have been described in neurodegenerative diseases, including AD and FTD. The Tau protein, which plays a central role in both diseases as hyperphosphorylated constituent of neurofibrillary tangles, is important for regulating microtubules, the intracellular transport tracks. Transport alterations appear to occur early in the cascade of neurological damage, long before neurons die. Therefore, axonal transport represents a promising drug target for neurodegenerative diseases. Mechanisms of axonal transport have been mainly studied in cultured neurons. While this approach has greatly expanded our knowledge of transport phenomena, it is also limited, as in vitro systems neither fully reflect the geometrical complexity of neurons in vivo and their interplay with other cells, nor easily mimic processes related to human disease. More intact preparations derived from invertebrates or mice circumvented some of these problems, but more versatile systems that allow studying axonal transport in vivo, together with screening for modulators, are still needed. Methods: We addressed this by developing an in vivo ‘tool box’ to study axonal transport in optically transparent and genetically accessible zebrafish. We have established the characteristics of mitochondrial transport in individually labelled neurons. Furthermore, we generated transgenic ‘mito-fish’ with fluorescently labelled neuronal mitochondria and performed quantitative measurements of mitochondrial density and transport in normal and compoundtreated animals. Results: As ‘proof-of-principle’, we crossed the ‘mitofish’ to a previously developed zebrafish tauopathy-model, which recapitulates key pathological features of FTD, including hyperphosphorylation, tangle formation and cell death (Paquet et al. 2009). Tau-transgenic fish show a prominent reduction in mitochondrial density and movements in peripheral neurons, which can be rescued by over-expression of MARK, a kinase that regulates Tau binding to microtubules. Conclusions: These approaches allow imaging transport of mitochondria in intact animals with high efficiency, and relating transport to the physiology of complete neurons in their natural habitat. Furthermore, ‘mito-fish’ are useful tools to assess mechanisms of transport disruptions and screen for modulating compounds.
S87
^ Abeta1-40/42 + CFA/IFA (n KLH+ CFA/IFA (n ¼ 8 Tg, 6 WT), 100 Amg ¼ 5 Tg, 8 WT), or vehicle (CFA/IFA; n ¼ 5 Tg, 6 WT) on days 0, 14, and 28 and monthly thereafter. Plasma was collected periodically. Splenocytes were cultured and restimulated with various Abeta peptides. Brains were examined pathologically and by ELISA. Results: J20tg and WT mice generated high antibody levels against the immunizing peptides. Antibodies from mice immunized with Abeta pE3-9 recognized Abeta pE340, 3-9, and 3-13 peptides; splenocytes proliferated upon restimulation with Abeta pE3-9/KLH, KLH, pE3-40and pE3-9 but not Abeta1-40/42 peptides. Antibodies from mice immunized with Abeta1-40/42 recognized Abeta1-40, 1-7 and 1-15 peptides; splenocytes proliferated upon restimulation with Abeta1-40/42 but not the Abeta pE3 peptides or KLH. IgG1 was the dominant isotype for both vaccines. Hippocampal fibrillar plaques were strongly reduced by immunization with Abeta pE3-9 (65%, p ¼ 0.05) and Abeta1-40/42 (69%, p ¼ 0.06) in the J20 tg mice compared to vehicle controls. AbetaX-42 levels were reduced in guanidine HCL brain homogenates of mice immunized by Abeta pE3-9 (28%, p ¼ 0.06) and Abeta1-40/42 (74%, p ¼ 0.002). Abeta pE3 levels were reduced by both vaccines but only reached significance in Abeta1-40/42 immunized mice. Both vaccines significantly increased plasma AbetaX-40 and lowered gliosis. Microhemorrhage was not observed, possibly due to low vascular amyloid at 14 months of age Conclusions: PyroGlu-3 Abeta is a strong immunogen that generates antibodies that lower plaque burden in APP tg mouse brain. Selective removal of toxic Abeta pE3-X may be beneficial for AD.
S1-02-06
TRANSGENIC MODELS TO ADDRESS TOXICITY
Kun Ping Lu, Harvard Medical School, Boston, Mass., United States. Abstract: not available.
SUNDAY, JULY 17, 2011 SYMPOSIA S1-03 PREVENTION OF DEMENTIA S1-03-01
EXERCISE, COGNITION AND THE BRAIN
Arthur Kramer, University of Illinois at Urbana-Champaign, Urbana-Champaign, Ill., United States. S1-02-05
ACTIVE IMMUNIZATION AGAINST PYROGLU-3 Ab GENERATES HIGH ANTIBODY TITERS AND LOWERS PLAQUE BURDEN IN APP TRANSGENIC MICE
Cynthia A Lemere1, Jeffrey Frost2, Martin Kleinschmidt3, Bin Liu2, Qiaoqiao Shi2, Hans-Ulrich Demuth4, Stephan Schilling5, 1Brigham & Women’s Hospital; Harvard Medical School, Boston, Massachusetts; 2 Brigham & Women’s Hospital, Boston, Massachusetts; 3Probiodrug AG, Halle (Saale); 4Probiodrug AG, Halle (Saale); 5Probiodrug AG, Halle. Background: Pyroglutamate-3 Abeta (Abeta pE3-X) is an N-terminally modified Abeta species that is abundant in human plaques and vascular amyloid. Abeta pE3 aggregates quickly, is resistant to degradation, and is neurotoxic. Abeta pE3 is present in AD-like transgenic (tg) mouse models, albeit at lower levels than Abeta1-X. In this pilot study, we compared the immune response and pathological efficacy of active immunization with Abeta pE3-9 vs. Abeta1-40/42 in APP tg and WT mice. Methods: J20 APP tg mice and WT mice of the same strain (B6D2F1) were actively immunized from 6 ^ Abeta pE3-9/ to 14 mo of age. Mice were given i.p. injections of 100 Amg
Background: Populations throughout the industrialized world are becoming increasing sedentary as a result of the changing nature of work and leisure activities. As a result of these societal changes increases in diseases such as hypertension, diabetes, osteoporosis, and forms of cancer are increasing. Physical activity serves to reduce susceptibility to these diseases. However, increased physical activity also has direct and relatively rapid effects on cognition and brain health. Such results have now been reported, over the course of several decades, in animal studies of physical activity. In my presentation I will review research conducted in our laboratory, and the field in general, which has examined the extent to which fitness training enhances cognition and brain structure and function of older adults. The presentation will cover both cross-sectional and intervention studies of fitness differences and fitness training. Studies which assess cognition via both behavioral measures and non-invasive neuroimaging measures, such as magnetic resonance imaging, functional magnetic resonance imaging, eventrelated brain potentials, and the event-related optical signal, will be reviewed and discussed. Finally, I will explore the gaps in the human and animal literature on cognitive and brain health and the manner in which they can be addressed in future research.