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Analysis of the cell distribution of endogenous murine leukemia virus in the brains of SAMRI and SAMP8 mice
Poster
Presentation:
Animal
Models I
REGIONAL ANALYSIS OF ABNORMAL TAU 1771BUTION IN PROGRESSIVE SUPRANUCLEAR
s17
PROTEIN PALSY
DISTRI-
OF THE CELL DISTRIBUTION OF ENDOGENOUS 1791ANALYSIS MURINE LEUKEMIA VIRUS IN THE BRAINS OF SAMRl AND SAMP8 MICE
Victoria A Zhukarevu, Univ of Pennsylvania, Philadelphia, PA: Steven S -M Chin, Columbus Univ. New York, NY; John .I Kulstad, John Q Trojanowski, Virginia M -Y Lee, Clniv of Pennsylvunia, Philadelphia, PA Progressive supranuclear palsy (PSP) is an atypical Parkinsonism with the dementia in the subset of cases. Neuropathologicaly, PSP is characterized by neuronal loss, gliosis and tau immunoreactive neurotibrillary tangles (NFT) in a specific brain regions i.e., basal ganglia, brain stem, and cerebellum, and to a lesser extend, in the perirhinal, inferior temporal and prefrontal cortex. Abnormal tau immunoreactive inclusions was are also detected in glial cells. The electrophoretic profile of pathological tau proteins in PSP is well characterized as a doublet of 64 kD and 69 kD recognized by anti-&w phosphorylatlon dependent antibody. We performed regional biochemical analysis of tau pathology distribution in six sporadic cases of PSP and two front”-temporal dementia linked to chromosome 17 (FIDP-17) cases with a clinical prese&tion of PSP. Quantitative Western blot analysis using phosphorylation dependent (PHFl) and phosphorylation independent (17026) antibody revealed the increasing gradient of pathological tau from cortical gray and white matter to cerebellum and bw al ganglia variable between different sporadic PSP cases. This variability may well reflect the progression and the severity of the disease. The similar pattern of the distribution of abnormal tau found in the two FTDP-17 cases with clinical symptoms of PSP suggests the possibility that pathology in thew two categories develops by a common mechanism.
Poster Presentation: IBUPROFEN 1781THOLOGY
Byung-hoon Jeong, Jae-Kwang Jin, Eun-Kynung Choi, Hallym Univ, Chuncheon South Korea; H. C Meeker, NYS lnst for Basrc Research in Developrnenml Disabilities, New York, NY; Christine A Kozak, NIH, Bethesda, MA: Richard I Carp, NYS lnst.for B&c Rrsearch in Developmental Disabilities, New York, NY; Yang-Sun Kim, Hallym Univ, Chuncheon South Korea Much higher levels of ecotropic murine leukemia virus (E-MuLV) are seen in the brains of accelerated senescence-prone (SAMPI) mice than in those of an accelerated senescence-resistant (SAMRI) strain. In order to investigate the cellular localization of E-MuLV in SAMP8, we determined the types of cells that express E-MuLV antigen in striatum, brainstem, hippocampus and cerebellum of SAMRl and SAMPS by immunohistochemistry. E-MuLV antigen was seen in the neurons and astrocytes of brain regions of SAMPX, but not in those of SAMR I. In SAMPS, immunoreactivity of glial fibrillary acidic protein (GFAP) was significantly enhanced. Vacuolation was detected only in brains of SAMPS, not in SAMRI, Furthermore, we analyzed the gene expression of mRNA of E-MuLV in the brains of SAMRI and SAMPB by RT-PCR and found that E-MuLV mRNA was expressed at a high level m SAMPI mice compared to SAMRl mice. Automatic DNA sequencing of a 605.base sequence in the envelop coding region of the SAMP8-derived virus mRNA (RT transcribed) was identical to the same region of the AKV MuLV proviral gene. A restriction map of the SAMP8 isolate showed no differences from that of AKV MuLV. These results suggest that E-MuLV generated from endogenous AKV provims replicates in neurons and astrocytea in the brains of SAMP8 and could cause the neuronal cell loss, astrocytosis, and vacuolation been in this mouse strain.
Animal Models I
REDUCES INFLAMMATION AND PLAQUE PAIN A MOUSE MODEL FOR ALZHEIMER’S DISEASE
Giselle P Lim. Fusheng Yang, Teresa Chu, Ping-ping Chen, Walter K Beech, Univ of CA, LOS Angeles and VAGLAHS, Sepulveda. CA; Oliver U&da, Univ of Los Angeles and VAGDLHS, Sepulveda, CA: Sally A Frautschy. Greg M Cole, UCLA and VAMC GRECC, Sepulvedu, CA Activated ghal cells and increased expression of cytokines and complement factors are associated with amyloid deposits and are consistent with a chronic inflammatory response in AD brain. More than 20 epidemiological studies indicate a reduced risk for Alzheimer’s disease associated with nonsteroldal anti-inflammatory drugs (NSAIDs) use, with ibuprofen being the most commonly used NSAID (50% of wbjects). These studies raise a question about the impact of NSAIDs, particularly ibuprofen, on the control of CNS inflammation and AD pathology. We tested the Impact of chronic oral ibuprofen in an amyloid precursor protein (HuAPPsw) transgenic mouse model for AD. This mice display widespread microglial activation, age-related hippocampal and neocortical amyloid deposits, and dystrophic neurites. Ten month old Tg (+) and (-) mice were fed chow containing 375 ppm ibuprofen for 6 months before they were sacrificed. ELISA and immunoblotting revealed a significant reduction (68.76%) in interleukin-lp and GFAP levels in several CNS regions of ibuprofen-treated mice. Ibuprofen decreased the percent area of antiphosphotyrosine-labeled microglia by 29.3% (P
MODIFICATION OF CYSTEINE PROTEASES AND AMPA RECEPTOR SUBUNITS BY THE LIPID PEROXIDATION PRODUCT 4HYROXYNONENAL: IMPACT ON NEURONAL APOPTOSIS AND NECROSIS S. L Ghan, C. Lu, M. P Mattson. National Institute on Aging, Baltimore, MD 4.hydroxynonenal (HNE), ir a lipid-peroxidation product that haa been shown to be elevated in Alzheimer’s brain tissues, can induce neuronal apoptosis at low concentrations and necrosis at higher concentrations. In order to understand the mechanism responsible for the switch in mode of cell death, we employed electrophysiological, calcium imaging, and biochemical analyses of cultured rat hippocampal neurons exposed to HNE. HNE caused a dose-dependent increase in AMPA current and AMPA receptor-mediated calcium influx, which was attenuated by the AMPA receptor antagonist CNQX. Immunoblot analysis showed no increase, but rather a time-dependent decrease, in AMPA receptor subunit levels following exposure to low doses of HNE (2-5 PM). The increase in channel actwity was partly attributed to direct binding of HNE to AMPA receptor subunits. High doses of HNE (20 PM) that induced necrosis had no effect on levels of AMPA receptor subunits. Immunoprecipitation and immunoblot studies using extracts from cortical neurons exposed to HNE showed direct binding of HNE to caspases and calpains, two major types of death-related cysteine proteases. Using fluorogenic pseudosubstrates and antibodies against specific breakdown products of calpain and caspase substrates, we demonstrated that activity of these proteases 1s reduced in situ in neurons exposed to HNE. Pretreatment of neurons with specific calpain and caspase inhibitors enhanced calcium responses to AMPA and sensitized neurons to necrotic cell death following exposure to low doses of HNE. Collectively, these data suggest that the switch in mode of cell death is directly attributable to the extent of calcium influx, which is determined in part by a direct effect of HNE on AMPA receptor function, and by indirectly inhibitmg AMPA receptor degradation as the result of cysteine pro&e inhibition. Effects of HNE on glutamate receptors and cyst&e proteases may contnbute to the neurodegenerative proces~ in AD.
Presentation:
Animal
Models I
REGIONAL ANALYSIS OF ABNORMAL TAU 1771BUTION IN PROGRESSIVE SUPRANUCLEAR
s17
PROTEIN PALSY
DISTRI-
OF THE CELL DISTRIBUTION OF ENDOGENOUS 1791ANALYSIS MURINE LEUKEMIA VIRUS IN THE BRAINS OF SAMRl AND SAMP8 MICE
Victoria A Zhukarevu, Univ of Pennsylvania, Philadelphia, PA: Steven S -M Chin, Columbus Univ. New York, NY; John .I Kulstad, John Q Trojanowski, Virginia M -Y Lee, Clniv of Pennsylvunia, Philadelphia, PA Progressive supranuclear palsy (PSP) is an atypical Parkinsonism with the dementia in the subset of cases. Neuropathologicaly, PSP is characterized by neuronal loss, gliosis and tau immunoreactive neurotibrillary tangles (NFT) in a specific brain regions i.e., basal ganglia, brain stem, and cerebellum, and to a lesser extend, in the perirhinal, inferior temporal and prefrontal cortex. Abnormal tau immunoreactive inclusions was are also detected in glial cells. The electrophoretic profile of pathological tau proteins in PSP is well characterized as a doublet of 64 kD and 69 kD recognized by anti-&w phosphorylatlon dependent antibody. We performed regional biochemical analysis of tau pathology distribution in six sporadic cases of PSP and two front”-temporal dementia linked to chromosome 17 (FIDP-17) cases with a clinical prese&tion of PSP. Quantitative Western blot analysis using phosphorylation dependent (PHFl) and phosphorylation independent (17026) antibody revealed the increasing gradient of pathological tau from cortical gray and white matter to cerebellum and bw al ganglia variable between different sporadic PSP cases. This variability may well reflect the progression and the severity of the disease. The similar pattern of the distribution of abnormal tau found in the two FTDP-17 cases with clinical symptoms of PSP suggests the possibility that pathology in thew two categories develops by a common mechanism.
Poster Presentation: IBUPROFEN 1781THOLOGY
Byung-hoon Jeong, Jae-Kwang Jin, Eun-Kynung Choi, Hallym Univ, Chuncheon South Korea; H. C Meeker, NYS lnst for Basrc Research in Developrnenml Disabilities, New York, NY; Christine A Kozak, NIH, Bethesda, MA: Richard I Carp, NYS lnst.for B&c Rrsearch in Developmental Disabilities, New York, NY; Yang-Sun Kim, Hallym Univ, Chuncheon South Korea Much higher levels of ecotropic murine leukemia virus (E-MuLV) are seen in the brains of accelerated senescence-prone (SAMPI) mice than in those of an accelerated senescence-resistant (SAMRI) strain. In order to investigate the cellular localization of E-MuLV in SAMP8, we determined the types of cells that express E-MuLV antigen in striatum, brainstem, hippocampus and cerebellum of SAMRl and SAMPS by immunohistochemistry. E-MuLV antigen was seen in the neurons and astrocytes of brain regions of SAMPX, but not in those of SAMR I. In SAMPS, immunoreactivity of glial fibrillary acidic protein (GFAP) was significantly enhanced. Vacuolation was detected only in brains of SAMPS, not in SAMRI, Furthermore, we analyzed the gene expression of mRNA of E-MuLV in the brains of SAMRI and SAMPB by RT-PCR and found that E-MuLV mRNA was expressed at a high level m SAMPI mice compared to SAMRl mice. Automatic DNA sequencing of a 605.base sequence in the envelop coding region of the SAMP8-derived virus mRNA (RT transcribed) was identical to the same region of the AKV MuLV proviral gene. A restriction map of the SAMP8 isolate showed no differences from that of AKV MuLV. These results suggest that E-MuLV generated from endogenous AKV provims replicates in neurons and astrocytea in the brains of SAMP8 and could cause the neuronal cell loss, astrocytosis, and vacuolation been in this mouse strain.
Animal Models I
REDUCES INFLAMMATION AND PLAQUE PAIN A MOUSE MODEL FOR ALZHEIMER’S DISEASE
Giselle P Lim. Fusheng Yang, Teresa Chu, Ping-ping Chen, Walter K Beech, Univ of CA, LOS Angeles and VAGLAHS, Sepulveda. CA; Oliver U&da, Univ of Los Angeles and VAGDLHS, Sepulveda, CA: Sally A Frautschy. Greg M Cole, UCLA and VAMC GRECC, Sepulvedu, CA Activated ghal cells and increased expression of cytokines and complement factors are associated with amyloid deposits and are consistent with a chronic inflammatory response in AD brain. More than 20 epidemiological studies indicate a reduced risk for Alzheimer’s disease associated with nonsteroldal anti-inflammatory drugs (NSAIDs) use, with ibuprofen being the most commonly used NSAID (50% of wbjects). These studies raise a question about the impact of NSAIDs, particularly ibuprofen, on the control of CNS inflammation and AD pathology. We tested the Impact of chronic oral ibuprofen in an amyloid precursor protein (HuAPPsw) transgenic mouse model for AD. This mice display widespread microglial activation, age-related hippocampal and neocortical amyloid deposits, and dystrophic neurites. Ten month old Tg (+) and (-) mice were fed chow containing 375 ppm ibuprofen for 6 months before they were sacrificed. ELISA and immunoblotting revealed a significant reduction (68.76%) in interleukin-lp and GFAP levels in several CNS regions of ibuprofen-treated mice. Ibuprofen decreased the percent area of antiphosphotyrosine-labeled microglia by 29.3% (P
MODIFICATION OF CYSTEINE PROTEASES AND AMPA RECEPTOR SUBUNITS BY THE LIPID PEROXIDATION PRODUCT 4HYROXYNONENAL: IMPACT ON NEURONAL APOPTOSIS AND NECROSIS S. L Ghan, C. Lu, M. P Mattson. National Institute on Aging, Baltimore, MD 4.hydroxynonenal (HNE), ir a lipid-peroxidation product that haa been shown to be elevated in Alzheimer’s brain tissues, can induce neuronal apoptosis at low concentrations and necrosis at higher concentrations. In order to understand the mechanism responsible for the switch in mode of cell death, we employed electrophysiological, calcium imaging, and biochemical analyses of cultured rat hippocampal neurons exposed to HNE. HNE caused a dose-dependent increase in AMPA current and AMPA receptor-mediated calcium influx, which was attenuated by the AMPA receptor antagonist CNQX. Immunoblot analysis showed no increase, but rather a time-dependent decrease, in AMPA receptor subunit levels following exposure to low doses of HNE (2-5 PM). The increase in channel actwity was partly attributed to direct binding of HNE to AMPA receptor subunits. High doses of HNE (20 PM) that induced necrosis had no effect on levels of AMPA receptor subunits. Immunoprecipitation and immunoblot studies using extracts from cortical neurons exposed to HNE showed direct binding of HNE to caspases and calpains, two major types of death-related cysteine proteases. Using fluorogenic pseudosubstrates and antibodies against specific breakdown products of calpain and caspase substrates, we demonstrated that activity of these proteases 1s reduced in situ in neurons exposed to HNE. Pretreatment of neurons with specific calpain and caspase inhibitors enhanced calcium responses to AMPA and sensitized neurons to necrotic cell death following exposure to low doses of HNE. Collectively, these data suggest that the switch in mode of cell death is directly attributable to the extent of calcium influx, which is determined in part by a direct effect of HNE on AMPA receptor function, and by indirectly inhibitmg AMPA receptor degradation as the result of cysteine pro&e inhibition. Effects of HNE on glutamate receptors and cyst&e proteases may contnbute to the neurodegenerative proces~ in AD.