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Activation and alteration of lysosomes in multiple system atrophy
P174 P1-191
Poster Presentations: P1 NEUROPROTECTIVE EFFECT OF FERMENTED LAMINARIA JAPONICA ON TMT-INDUCED COGNITIVE DEFICIT IN RATS
Hyun Jung Park1, Hyun Soo Shim1, Insop Shim2, 1Kyung Hee University, Seoul, South Korea; 2Kyung Hee University/Acupuncture and Meridian Science Research Center and College of Oriental Medicine, Seoul, South Korea. Background: Sea tangle (Laminaria japonica), an edible brown seaweed, has long been utilized since ancient times as an important food resource to promote maternal health in Pacific and Asian countries. Methods: The rats were administered saline or LJ (LJ 50, 100 and 200 mg/kg, p.o.) daily for 21 days. The cognitive improving efficacy of LJ on the amnesic rats, which was induced by TMT, was investigated by assessing the Morris water maze test and by performing acetylchoine transferase (ChAT) immunohistochemistry. In the in vitro study, in order to confirm the underlying mechanisms of the memory enhancing effects of LJ, we assessed the neurite outgrowth of PC12 cells and expression of amyloid beta in the H4 cells. Results: The rats with TMT injection showed impaired learning and memory of the tasks and treatment with LJ produced a significant improvement of the escape latency to find the platform in the Morris water maze at the 2nd day compared to that of the control group (at the 4th day, P <0.05). In the retention test, the LJ treated group showed increased time spent around the platform compared to that of the control group. Consistent with the behavioral data, LJ treated group significantly alleviated the loss of ChAT-ir neurons in the hippocampus compared to that of the control group. Treatment with LJ significantly promoted neurite outgrowth of the PC12 cells and reduced the expression of amyloid-beta in H4 cells, as compared to that of the controls. Conclusions: These results suggest that LJ may be useful for improving the cognitive function via regulation of cholinergic marker enzyme activity and neurite outgrowth and neurotoxicity. P1-192
ACTIVATION AND ALTERATION OF LYSOSOMES IN MULTIPLE SYSTEM ATROPHY
Kouki Makioka1, Tsuneo Yamazaki2, Koichi Okamoto2, 1Gunma University Graduate School of Medicine, Maebahi, Japan; 2Gunma University, Maebashi, Japan. Background: Accumulation and deposition of misfolded proteins are common signs of neurodegenerative diseases. The regional distribution in the brain and the composition of protein aggregates are different in each neurodegenerative disease. Recently, dysfunction of protein degradation systems was proposed as one of the causes of the accumulation of these aberrant proteins. A normal balance between the formation and degradation of cellular proteins is required for cell survival. The autophagy-lysosome system is mainly responsible for the nonselective degradation of proteins. Lysosomal alteration is observed in many neurodegenerative diseases, even if lysosomal disturbance does not represent the direct cause of disease. This indicates that lysosomes play a role in neurodegeneration. Multiple system atrophy (MSA) is a sporadic neurodegenerative disorder. Its histopathological features include glial cytoplasmic inclusions (GCIs), which contain 6 -synuclein as the main component. It has been reported recently that a -synuclein is also degraded via the autophagy-lysosomal pathway. The participation of the autophagy-lysosome pathway in MSA pathogenesis has not been reported. Methods: Brain tissues were obtained from nine subjects (average age, 60.6 years; five males and four females) who had a neuropathologically confirmed diagnosis of MSA, and from five control subjects with no signs of neurodegeneration. Five-micrometer-thick sections of formalin-fixed, paraffin-embedded tissues from the pons were immunostained with the appropriate antibodies using the streptavidin-biotin method. To assess the colocalization of lysosomal proteins with GCI-associated proteins and microglial marker, we performed double immunohistochemistry using fluorescent secondary antibodies. Results: A robust increase in the expression, and alteration in the morphology and distribution of lysosomal-protein-positive structures were observed in MSA
brains. Double immunohistochemistry demonstrated that lysosomal markers did not colocalize mainly with GCIs, but colocalized with a microglial marker. Conclusions: These immunohistochemical signatures suggest that lysosomes are activated in microglia during the disease process and play a pivotal role in the pathology of MSA.
P1-193
AUTOIMMUNIZATION LEADS TO ELEVATED ANTI-BETA-AMYLOID ANTIBODIES IN DOWN SYNDROME (DS)
Antonia Coppus1, Marc Weksler2, Paul Szabo3, Norman Relkin3, 1Erasmus University Medical Center, Rotterdam, Netherlands; 2Weill Cornell Medical College, Tenafly, NJ, New Jersey, United States; 3Weill Cornell Medical College, New York, New York, United States. Background: Individuals with Down’s Syndrome (DS) overproduce beta amyloid throughout life as a consequence of triplication of the APP gene. This would be expected to lead to immune tolerance, but endogenous anti-amyloid monomer antibodies have been reported to be elevated in DS patients. In contrast, elderly normals and AD patients have reduced levels of anti-monomer antibodies. We examined anti-amyloid antibody expression in Downs to better understand its relationship to the development of dementia in DS. Methods: 178 plasma specimens from DS patients were obtained under an IRB-approved protocol at Erasmus University (Rotterdam). IgG samples were isolated by Thiophilic absorbant chromatography and ELISAs for anti-amyloid antibodies were performed (Szabo et al., 2010). Antibody affinities were determined by SPR using SensiQ (ICX-Nomadics, Oklahoma). Antidiphtheria antibody titers were obtained by Viracor-IBT Laboratories as a proxy for immune competence. Results: Compared to anti-abeta antibodies from normal human donors (IVIG), the DS patients had 6.9 fold greater monomer, 2.8-fold greater anti-oligomer and 5-fold greater anti-fibril antibody titers. The dissociation constant (Kd) of N-terminal anti-monomer antibodies from DS probands was 520 nM compared to 2,280 nM for IVIg. Anti-monomer response correlated with the anti-diphtheria response in most DS patients. However, antioligomer response correlated with anti-diphtheria only in those individuals who had dementia or developed dementia during the study. No correlation was found between the anti-diphtheria titers and the anti-fibril response. Conclusions: DS probands produce relatively high levels of anti-amyloid monomer antibodies that have undergone immune maturation resulting in higher affinity. This is consistent with self-immunization against beta amyloid monomer being part of an auto-immune disturbance in DS. Anti-oligomer antibody expression may be more closely related to the development of dementia in Downs than either anti-monomer or anti-fibril antibody expression.
P1-194
NOVEL DRUG STRATEGIES AND MOLECULAR MECHANISMS OF LYCOPENE, FK-506 AND VENLAFAXINE AGAINST 3-NP INDUCED HUNTINGTON’S-LIKE SYMPTOMS IN RATS
Puneet Bansal1, Anil Kumar2, 1ISF College of Pharmacy, Barnala, India; Panjab University, Chandigarh, Chandigarh, India.
2
Background: Huntington’s disease (HD) is a progressive neurodegenerative disorder characterized by a degeneration of striatal neurons. Numbers of pathways are involved in the pathphysiology of this complex disease, so targeting one pathway may not be adequate drug therapy for HD. Here we tried to target different pathways with combination therapy like antioxidant and FK-506 to reduce the oxidative stress, antidepressants to abolish the psychiatric symptoms.The present study was designed to elucidate the neuroprotective drug strategies by combining different classes of drugs like antioxidant (lycopene), immunophilines (FK506), antidepressants (venlafaxine) and to explore their molecular mechanisms against 3-NP induced HD like symptoms. Methods: Experimental protocol includes systemic administration of 3-
Poster Presentations: P1 NEUROPROTECTIVE EFFECT OF FERMENTED LAMINARIA JAPONICA ON TMT-INDUCED COGNITIVE DEFICIT IN RATS
Hyun Jung Park1, Hyun Soo Shim1, Insop Shim2, 1Kyung Hee University, Seoul, South Korea; 2Kyung Hee University/Acupuncture and Meridian Science Research Center and College of Oriental Medicine, Seoul, South Korea. Background: Sea tangle (Laminaria japonica), an edible brown seaweed, has long been utilized since ancient times as an important food resource to promote maternal health in Pacific and Asian countries. Methods: The rats were administered saline or LJ (LJ 50, 100 and 200 mg/kg, p.o.) daily for 21 days. The cognitive improving efficacy of LJ on the amnesic rats, which was induced by TMT, was investigated by assessing the Morris water maze test and by performing acetylchoine transferase (ChAT) immunohistochemistry. In the in vitro study, in order to confirm the underlying mechanisms of the memory enhancing effects of LJ, we assessed the neurite outgrowth of PC12 cells and expression of amyloid beta in the H4 cells. Results: The rats with TMT injection showed impaired learning and memory of the tasks and treatment with LJ produced a significant improvement of the escape latency to find the platform in the Morris water maze at the 2nd day compared to that of the control group (at the 4th day, P <0.05). In the retention test, the LJ treated group showed increased time spent around the platform compared to that of the control group. Consistent with the behavioral data, LJ treated group significantly alleviated the loss of ChAT-ir neurons in the hippocampus compared to that of the control group. Treatment with LJ significantly promoted neurite outgrowth of the PC12 cells and reduced the expression of amyloid-beta in H4 cells, as compared to that of the controls. Conclusions: These results suggest that LJ may be useful for improving the cognitive function via regulation of cholinergic marker enzyme activity and neurite outgrowth and neurotoxicity. P1-192
ACTIVATION AND ALTERATION OF LYSOSOMES IN MULTIPLE SYSTEM ATROPHY
Kouki Makioka1, Tsuneo Yamazaki2, Koichi Okamoto2, 1Gunma University Graduate School of Medicine, Maebahi, Japan; 2Gunma University, Maebashi, Japan. Background: Accumulation and deposition of misfolded proteins are common signs of neurodegenerative diseases. The regional distribution in the brain and the composition of protein aggregates are different in each neurodegenerative disease. Recently, dysfunction of protein degradation systems was proposed as one of the causes of the accumulation of these aberrant proteins. A normal balance between the formation and degradation of cellular proteins is required for cell survival. The autophagy-lysosome system is mainly responsible for the nonselective degradation of proteins. Lysosomal alteration is observed in many neurodegenerative diseases, even if lysosomal disturbance does not represent the direct cause of disease. This indicates that lysosomes play a role in neurodegeneration. Multiple system atrophy (MSA) is a sporadic neurodegenerative disorder. Its histopathological features include glial cytoplasmic inclusions (GCIs), which contain 6 -synuclein as the main component. It has been reported recently that a -synuclein is also degraded via the autophagy-lysosomal pathway. The participation of the autophagy-lysosome pathway in MSA pathogenesis has not been reported. Methods: Brain tissues were obtained from nine subjects (average age, 60.6 years; five males and four females) who had a neuropathologically confirmed diagnosis of MSA, and from five control subjects with no signs of neurodegeneration. Five-micrometer-thick sections of formalin-fixed, paraffin-embedded tissues from the pons were immunostained with the appropriate antibodies using the streptavidin-biotin method. To assess the colocalization of lysosomal proteins with GCI-associated proteins and microglial marker, we performed double immunohistochemistry using fluorescent secondary antibodies. Results: A robust increase in the expression, and alteration in the morphology and distribution of lysosomal-protein-positive structures were observed in MSA
brains. Double immunohistochemistry demonstrated that lysosomal markers did not colocalize mainly with GCIs, but colocalized with a microglial marker. Conclusions: These immunohistochemical signatures suggest that lysosomes are activated in microglia during the disease process and play a pivotal role in the pathology of MSA.
P1-193
AUTOIMMUNIZATION LEADS TO ELEVATED ANTI-BETA-AMYLOID ANTIBODIES IN DOWN SYNDROME (DS)
Antonia Coppus1, Marc Weksler2, Paul Szabo3, Norman Relkin3, 1Erasmus University Medical Center, Rotterdam, Netherlands; 2Weill Cornell Medical College, Tenafly, NJ, New Jersey, United States; 3Weill Cornell Medical College, New York, New York, United States. Background: Individuals with Down’s Syndrome (DS) overproduce beta amyloid throughout life as a consequence of triplication of the APP gene. This would be expected to lead to immune tolerance, but endogenous anti-amyloid monomer antibodies have been reported to be elevated in DS patients. In contrast, elderly normals and AD patients have reduced levels of anti-monomer antibodies. We examined anti-amyloid antibody expression in Downs to better understand its relationship to the development of dementia in DS. Methods: 178 plasma specimens from DS patients were obtained under an IRB-approved protocol at Erasmus University (Rotterdam). IgG samples were isolated by Thiophilic absorbant chromatography and ELISAs for anti-amyloid antibodies were performed (Szabo et al., 2010). Antibody affinities were determined by SPR using SensiQ (ICX-Nomadics, Oklahoma). Antidiphtheria antibody titers were obtained by Viracor-IBT Laboratories as a proxy for immune competence. Results: Compared to anti-abeta antibodies from normal human donors (IVIG), the DS patients had 6.9 fold greater monomer, 2.8-fold greater anti-oligomer and 5-fold greater anti-fibril antibody titers. The dissociation constant (Kd) of N-terminal anti-monomer antibodies from DS probands was 520 nM compared to 2,280 nM for IVIg. Anti-monomer response correlated with the anti-diphtheria response in most DS patients. However, antioligomer response correlated with anti-diphtheria only in those individuals who had dementia or developed dementia during the study. No correlation was found between the anti-diphtheria titers and the anti-fibril response. Conclusions: DS probands produce relatively high levels of anti-amyloid monomer antibodies that have undergone immune maturation resulting in higher affinity. This is consistent with self-immunization against beta amyloid monomer being part of an auto-immune disturbance in DS. Anti-oligomer antibody expression may be more closely related to the development of dementia in Downs than either anti-monomer or anti-fibril antibody expression.
P1-194
NOVEL DRUG STRATEGIES AND MOLECULAR MECHANISMS OF LYCOPENE, FK-506 AND VENLAFAXINE AGAINST 3-NP INDUCED HUNTINGTON’S-LIKE SYMPTOMS IN RATS
Puneet Bansal1, Anil Kumar2, 1ISF College of Pharmacy, Barnala, India; Panjab University, Chandigarh, Chandigarh, India.
2
Background: Huntington’s disease (HD) is a progressive neurodegenerative disorder characterized by a degeneration of striatal neurons. Numbers of pathways are involved in the pathphysiology of this complex disease, so targeting one pathway may not be adequate drug therapy for HD. Here we tried to target different pathways with combination therapy like antioxidant and FK-506 to reduce the oxidative stress, antidepressants to abolish the psychiatric symptoms.The present study was designed to elucidate the neuroprotective drug strategies by combining different classes of drugs like antioxidant (lycopene), immunophilines (FK506), antidepressants (venlafaxine) and to explore their molecular mechanisms against 3-NP induced HD like symptoms. Methods: Experimental protocol includes systemic administration of 3-