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Increased expression of S100β in brain of IM mice injected with 87V scrapie strain
FOURTH INTERNATIONAL CONFERENCE ON ALZHEIMER’S DISEASE
PET data indicate that the disease process, even at early stages, is more widespread in subjects heterozygous at polymorphic codOn 129 than in those homozygous. Therefore, it is possible that the polymorphism at codon 129 influences not only the duration of the disease but also the distribution of PrP-res and the topography of the lesions. Supported by AG-08012, AG-08155, AG-08992 and the Brkton Fund.
cortex as well as caudate nucleus in scrapie injected mouse. Further analysis of Northern blot showed the levels of S1008 mRNA were significantly elevated in injected mice brain compared to those of control mice. Taken together, our results suggest that SlOO8 may involve in formation of amyloid plaques on scrapie injected mice. Acknowledgement : This study was supported by a grant for genetic engineering from the Ministry of Education(92-131) of Korea.
364 GERSTMANN-STRAUSSLER-SCHEINKER DISEASE: GENEALOGICAL, ANATOMICAL AND IMMUNOHISTOCHEMISTRY STUDIES OF A FRENCH FAMILY. K. El Hachimi, G. Giaccone, F. Tagliavini, 0. Bugiani and I.-F. Foncin. Neurohistologie EPHE and U.106 INSERM, La SalpWere Paris France and Istituto Neurologico C. Besta Milan0 Italy. About 200 persons, 6 of whom were affected by Gerstmann-StrlusslerScheinker disease (GSS), are identified in the pedigree.The mutation has not yet been characterized.This family, originating from the north of France, is histopathologically characterized by lesions restrictively located to the cerebellar cortex, the molecular of which is moderately atrophic and gliotic, and contains numemus plaque-like formation without neuritic component and evidenced by PAS staining, but not by Congo red birefringence. However, ultrastructural study, on autopsy material, showeduni- or multicentric amyloid plaques made of radiating amyloid fibrillaty bundles. Numerous electron-dense granules and vesicles were seen between the bundles. Few degenerating neurites were seen around these plaques, but microglial cells were present. At the light microscope immunohistochemistry level, the plaques were selectively stained with anti-PrP, located exclusively througout the cerebellar layer. Immunoreactivity was also observed in Virchow-Robin spaces.The amyloid fibrils were selectively immunodecorated at the immuno-ultrastructural level. The plaque were associated with abundant microglia/macrophage cells evidenced by anti-fenitin immunostaining, but MHC class II (LN3) positive-microglia cells were absent. GFAP immunoreactivity was moderate. Immunoprocessing for Tau protein and ubiquitin revealed a few degenerating neurites. Amyloid deposits were not immunolabeled with antibodies against the Alzheimer disease B/A4 protein. No immunoreactive PrP-plaque were identified in other regions of cerebellum or cerebrum. The particular topography of lesions and the absence of other lesions lead us to compare this family with the 102 (Pro->Leu) PrP gene mutation observed in others families from different countries.
366 CYTOKINES, PROSTAGLANDINS AND LIPOCORTIN-1 ARE PRESENT IN THE BRAINS OF SCRAP&INFECTED MICE. A.E. Williams, A.-M. van Dam’, F. Berkenbosch’, P. Eikelenboomt and H. Fraser. AFRClMRC Neuropathogenesis Unit, Institute for Animal Health, Edinburgh, EH9 3JF, Scotland and Departments of ?? Pharmacology and tPsychiatrY, Free University, Amsterdam, Netherlands. The neuropathology of scrapie and Creutzfeldt-Jakob disease is cheracterised by chronic neurodegeneration with vacuolation of neurons end/or neuropil and amyloid plaque formation in some cases. A classical acute inflammatory response end immunopathology in the form of perivascular cuffs are absent. A modified, protease-resistant isoform (termed PrP”) of the host-encoded PrP protein accumulates in those areas of the brain showing vacuolation and plaques; astrocyte hypertrophy and proliferation and a marked microgliel activation are also present in these areas. The microglia show upregulation of leukocyte antigens with known functions in recruitment, phagocytosis (CR3), leukocyte activation (LCA) and antrgen presentatron lMHC III whrch suggests that the mrcrogliai response represents a modified inflammatory response. In the present studies, the presence of cytokinas, prostaglandins and lipocortin-1 was investigated in terminally-affected mice of four scrapie models, using immunocytochemical techniques. Two of these models show “estrocvte halos” of PrP staining end the other two PrP deposition mainly associated with neurons but also with occasional microglia. There was marked induction of interleukin-16, interleukin-6, tumour necrosis factor (I, prostaglandin IPG) Es, PGF,, and lipocortin-1. Although the distribution of immunoreactive cells differed between the various models, the pattern of staining within each model was similar for all factors examined and corresponded to areas of the brain showing vacuolation of neuronslneuropil, astrocytosis and microglial activation. The type of glial cell expressing these factors was determined by comparing the results of their immunolabelling with the morphology of GFAP (estrocvtic) and F4160 Imicroglial) immunoreactive cells. It has previously been proposed that cytokines influence or mediate abnormal APP processing and RA4 deposition in Alzheimer’sdisease. The findings reported here indicate that similar mechanisms may also be relevant in PrP processing and the pathogenesis of neurodegeneretion in scrapie.
365 INCREASED EXPRESSION OF SlOO8 IN BRAIN OF IM MICE INJECTED WITH 87V SCRAPIE STRAIN. Y.S. Kiml, E.K. Choil, H.M. Yang?. J. Kims, I.J. Yu3, D.R. Marshakd. R.I. Carps, and H.M. Wisniewskis. lKorea Institute of Gerontology, College of Med., Hallym Univ., Zollege of Med., Catholic Univ.. aIndustrial Health Research Institute. Korea Industrial Safety Corporation, Seoul, Korea, cold Spring Harbor Lab. NY, and sNew York State Institute for Basic Research in Developmental Disabilities, NY, USA. Recently the presence of increased levels of neurotrophic factors such as SlOO8 in Alzheimer’s disease(AD) is one possible explanation for the increased concentration of aggregates of overgrown neurites in neuritic plaques of AD. Little is known about the functions of SlOO8 in chronic neurodegenerative disease. To study a role for SlOO8 protein in formation of amyloid plaques, IM mice were injected with 87V scrapie strain and analyzed for the expression S1008. Microinjections were carried out under general anesthesia using a stereotaxic instrument. The injection volume was 5 II of 1% of 87V scrapie brain homogenate. The stereotaxic coordinate used for cerebral cortex was A +l .O. L +2.0. H +1.5. Clinical evaluation of motor coordination was monitored with using a grid apparatus. Brains of positive scored mice were removed and used for further analyses. The injected mice developed amyloid plaques in the regions of cerebral cortex, corpus callosum and hippocampus. The results of immunohistochemistry on scrapie injected brain showed increases of SlOO8 immunoreactivity in the regions of cerebral cortex, hippocampus. mesencephalon, and caudate nucleus compared to control mouse. In addition, glial fibrillary acidic protein (GFAP) immunoreactive cells were markedly increased in the cerebral
367 TRUNCATED FORMS OF PRION PROTEIN IN HUMAN BRAIN AND CELL LINES. S.G. Chen, R. Petersen, P. Parchi, L. Monari, W. Wang, P. Gambetti, L Gambetti. Division of Neuropathology, Case Western Reserve University, 2085 Adelbert Road, Cleveland, Ohio 44108 USA. Prion protein (PrP) is a glycoprotein attached to the cell membrane by a glycosyl-phosphatidylinoskol (GPI) anchor. Although its normal cellular function is not known, an aberrant isoform of PrP is a hallmark of prion diseases in humans and animals. We have characterized PrP from human brains and neuroblastoma cell lines. Using sequence-specific antibodies, we found that in addlion to full length PrP, a comparable amount of an N-terminal truncated form is present in cultured cells and in normal brains. As the full length PrP, most of the N-terminal truncated isoform is attached to the cell surface through a GPI anchor, since it is released from cells in culture as well as from brain membrane fractions after treatment with phosphatidylinositol-specific phospholipase C. Similar fragments were present in brains from cases of Creutzfeld-Jakob disease and Fatal Familial Insomnia but were not protease-resistant. Increased expression of the truncated PrP was observed in transfected cells overexpressing the PrP gene, indicating that the N-terminal truncated PrP derives from processing of full length PrP. Studies of events underlying processing of prion protein may help to elucidate the role of PrP in normal and pathological conditions. Supported by AG-08155 and AG-08992.
PET data indicate that the disease process, even at early stages, is more widespread in subjects heterozygous at polymorphic codOn 129 than in those homozygous. Therefore, it is possible that the polymorphism at codon 129 influences not only the duration of the disease but also the distribution of PrP-res and the topography of the lesions. Supported by AG-08012, AG-08155, AG-08992 and the Brkton Fund.
cortex as well as caudate nucleus in scrapie injected mouse. Further analysis of Northern blot showed the levels of S1008 mRNA were significantly elevated in injected mice brain compared to those of control mice. Taken together, our results suggest that SlOO8 may involve in formation of amyloid plaques on scrapie injected mice. Acknowledgement : This study was supported by a grant for genetic engineering from the Ministry of Education(92-131) of Korea.
364 GERSTMANN-STRAUSSLER-SCHEINKER DISEASE: GENEALOGICAL, ANATOMICAL AND IMMUNOHISTOCHEMISTRY STUDIES OF A FRENCH FAMILY. K. El Hachimi, G. Giaccone, F. Tagliavini, 0. Bugiani and I.-F. Foncin. Neurohistologie EPHE and U.106 INSERM, La SalpWere Paris France and Istituto Neurologico C. Besta Milan0 Italy. About 200 persons, 6 of whom were affected by Gerstmann-StrlusslerScheinker disease (GSS), are identified in the pedigree.The mutation has not yet been characterized.This family, originating from the north of France, is histopathologically characterized by lesions restrictively located to the cerebellar cortex, the molecular of which is moderately atrophic and gliotic, and contains numemus plaque-like formation without neuritic component and evidenced by PAS staining, but not by Congo red birefringence. However, ultrastructural study, on autopsy material, showeduni- or multicentric amyloid plaques made of radiating amyloid fibrillaty bundles. Numerous electron-dense granules and vesicles were seen between the bundles. Few degenerating neurites were seen around these plaques, but microglial cells were present. At the light microscope immunohistochemistry level, the plaques were selectively stained with anti-PrP, located exclusively througout the cerebellar layer. Immunoreactivity was also observed in Virchow-Robin spaces.The amyloid fibrils were selectively immunodecorated at the immuno-ultrastructural level. The plaque were associated with abundant microglia/macrophage cells evidenced by anti-fenitin immunostaining, but MHC class II (LN3) positive-microglia cells were absent. GFAP immunoreactivity was moderate. Immunoprocessing for Tau protein and ubiquitin revealed a few degenerating neurites. Amyloid deposits were not immunolabeled with antibodies against the Alzheimer disease B/A4 protein. No immunoreactive PrP-plaque were identified in other regions of cerebellum or cerebrum. The particular topography of lesions and the absence of other lesions lead us to compare this family with the 102 (Pro->Leu) PrP gene mutation observed in others families from different countries.
366 CYTOKINES, PROSTAGLANDINS AND LIPOCORTIN-1 ARE PRESENT IN THE BRAINS OF SCRAP&INFECTED MICE. A.E. Williams, A.-M. van Dam’, F. Berkenbosch’, P. Eikelenboomt and H. Fraser. AFRClMRC Neuropathogenesis Unit, Institute for Animal Health, Edinburgh, EH9 3JF, Scotland and Departments of ?? Pharmacology and tPsychiatrY, Free University, Amsterdam, Netherlands. The neuropathology of scrapie and Creutzfeldt-Jakob disease is cheracterised by chronic neurodegeneration with vacuolation of neurons end/or neuropil and amyloid plaque formation in some cases. A classical acute inflammatory response end immunopathology in the form of perivascular cuffs are absent. A modified, protease-resistant isoform (termed PrP”) of the host-encoded PrP protein accumulates in those areas of the brain showing vacuolation and plaques; astrocyte hypertrophy and proliferation and a marked microgliel activation are also present in these areas. The microglia show upregulation of leukocyte antigens with known functions in recruitment, phagocytosis (CR3), leukocyte activation (LCA) and antrgen presentatron lMHC III whrch suggests that the mrcrogliai response represents a modified inflammatory response. In the present studies, the presence of cytokinas, prostaglandins and lipocortin-1 was investigated in terminally-affected mice of four scrapie models, using immunocytochemical techniques. Two of these models show “estrocvte halos” of PrP staining end the other two PrP deposition mainly associated with neurons but also with occasional microglia. There was marked induction of interleukin-16, interleukin-6, tumour necrosis factor (I, prostaglandin IPG) Es, PGF,, and lipocortin-1. Although the distribution of immunoreactive cells differed between the various models, the pattern of staining within each model was similar for all factors examined and corresponded to areas of the brain showing vacuolation of neuronslneuropil, astrocytosis and microglial activation. The type of glial cell expressing these factors was determined by comparing the results of their immunolabelling with the morphology of GFAP (estrocvtic) and F4160 Imicroglial) immunoreactive cells. It has previously been proposed that cytokines influence or mediate abnormal APP processing and RA4 deposition in Alzheimer’sdisease. The findings reported here indicate that similar mechanisms may also be relevant in PrP processing and the pathogenesis of neurodegeneretion in scrapie.
365 INCREASED EXPRESSION OF SlOO8 IN BRAIN OF IM MICE INJECTED WITH 87V SCRAPIE STRAIN. Y.S. Kiml, E.K. Choil, H.M. Yang?. J. Kims, I.J. Yu3, D.R. Marshakd. R.I. Carps, and H.M. Wisniewskis. lKorea Institute of Gerontology, College of Med., Hallym Univ., Zollege of Med., Catholic Univ.. aIndustrial Health Research Institute. Korea Industrial Safety Corporation, Seoul, Korea, cold Spring Harbor Lab. NY, and sNew York State Institute for Basic Research in Developmental Disabilities, NY, USA. Recently the presence of increased levels of neurotrophic factors such as SlOO8 in Alzheimer’s disease(AD) is one possible explanation for the increased concentration of aggregates of overgrown neurites in neuritic plaques of AD. Little is known about the functions of SlOO8 in chronic neurodegenerative disease. To study a role for SlOO8 protein in formation of amyloid plaques, IM mice were injected with 87V scrapie strain and analyzed for the expression S1008. Microinjections were carried out under general anesthesia using a stereotaxic instrument. The injection volume was 5 II of 1% of 87V scrapie brain homogenate. The stereotaxic coordinate used for cerebral cortex was A +l .O. L +2.0. H +1.5. Clinical evaluation of motor coordination was monitored with using a grid apparatus. Brains of positive scored mice were removed and used for further analyses. The injected mice developed amyloid plaques in the regions of cerebral cortex, corpus callosum and hippocampus. The results of immunohistochemistry on scrapie injected brain showed increases of SlOO8 immunoreactivity in the regions of cerebral cortex, hippocampus. mesencephalon, and caudate nucleus compared to control mouse. In addition, glial fibrillary acidic protein (GFAP) immunoreactive cells were markedly increased in the cerebral
367 TRUNCATED FORMS OF PRION PROTEIN IN HUMAN BRAIN AND CELL LINES. S.G. Chen, R. Petersen, P. Parchi, L. Monari, W. Wang, P. Gambetti, L Gambetti. Division of Neuropathology, Case Western Reserve University, 2085 Adelbert Road, Cleveland, Ohio 44108 USA. Prion protein (PrP) is a glycoprotein attached to the cell membrane by a glycosyl-phosphatidylinoskol (GPI) anchor. Although its normal cellular function is not known, an aberrant isoform of PrP is a hallmark of prion diseases in humans and animals. We have characterized PrP from human brains and neuroblastoma cell lines. Using sequence-specific antibodies, we found that in addlion to full length PrP, a comparable amount of an N-terminal truncated form is present in cultured cells and in normal brains. As the full length PrP, most of the N-terminal truncated isoform is attached to the cell surface through a GPI anchor, since it is released from cells in culture as well as from brain membrane fractions after treatment with phosphatidylinositol-specific phospholipase C. Similar fragments were present in brains from cases of Creutzfeld-Jakob disease and Fatal Familial Insomnia but were not protease-resistant. Increased expression of the truncated PrP was observed in transfected cells overexpressing the PrP gene, indicating that the N-terminal truncated PrP derives from processing of full length PrP. Studies of events underlying processing of prion protein may help to elucidate the role of PrP in normal and pathological conditions. Supported by AG-08155 and AG-08992.