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c-Rel deficient mice, a mouse model of “spreading” PD-like pathology
Abstracts / Parkinsonism and Related Disorders 22 (2016) e149ee192
Objectives: In accordance with modern basal ganglia model, genetic code alteration responsible for formation of corticobasal circuit should lead to changes of movement and behavior. Methods: Within 3 years, line of outbred mice, which were inbreed for 8 generations (16 mutant mice and 10 controls) was studied. Weight monitoring and open field test were done. Assessment of social behavior was analyzed by numbers of newborn mice. Quality of mutation was determined by direct and backcrossing in 3 generations. Results: Selection showed that mutation is spontaneous and recessive linked to X chromosome. Open field test showed that 2 out of 16 mutants were able to get out (6.3 min, 1.4 min control). Control group showed normal social behavior, number of mice born was 7 in average, average generation time was 78 days. Mutants have 6 mice, but as females didn't express parental instinct, number of two week age mice reached 0. All mutants were grown up by either control mice or carriers of mutant gene. Generation time e 147d. Weight monitoring isn't informative. Phenotype is formed by 4th week of development with progresses to the second month, manifested by stereotypical circular movements in both directions, and inability to walk in straight line, the diameter of the circle depends on the degree of animals' anxiety. Calm animal perform pendulum motion with linear trajectory. Frightened animal rotates around gravity axis. Conclusions: Considering the phenotype of mutation, we assumed a defect in the structure of reelin. This mutation would be good model for stereotyping movement disorders. P 6.014. GENETIC MODELS OF PARKINSON'S DISEASE Moein Dehbashi 1, 3, Maedeh Rouigari 2,3. 1 Genetics Division, Biology Department, Faculty of Science, University of Isfahan, Isfahan, Iran; 2 Biology Department, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran; 3 Isfahan Neurosciences Research Center, Al-Zahra Hospital, Isfahan University of Medical Sciences, Isfahan, Iran Objectives: The disappointing outcomes obtained so far by modelling the genetics of Parkinson's disease (PD) in mice have motivated interest in developing alternative genetic models in multicellular model organisms. Methods: Here we will focus on the fruit fly (Drosophila melanogaster), the nematode Caenorhabditis Elegans and the zebrafish (Danio Rerio) as the PD models. Results: The fruit fly (D. melanogaster), the nematode C. Elegans and the zebrafish (Danio Rerio) exhibit some clear advantages over rodents in terms of the relative ease with which the genome can be manipulated and of the much reduced costs involved in the development of genetic models of PD, but of course, their face validity is limited by the nature of the ‘symptoms’ these species present with. Conclusions: Given that these models are in much earlier stages of development, they have yet to play a role in drug discovery for PD; however, they may prove invaluable in the future development of diseasemodifying strategies that have so far yielded little success in clinical efforts. References: 1. Dawson TM, Ko HS, Dawson VL. Genetic animal models of Parkinson's disease. Neuron 2010;66:646-661. 2. Duty S, Jenner P. Animal models of Parkinson's disease: a source of novel treatments and clues to the cause of the disease. Br J Pharmacol 2011;164:1357-1391. 3. Le W, Sayana P, Jankovic J. Animal models of Parkinson's disease: a gateway to therapeutics? Neurotherapeutics 2014;11:92-110. P 6.015. PROGRESSIVE AXONAL DEGENERATION OF THE STRIATONIGRAL DOPAMINERGIC NEURONS IN CALCIUM-INDEPENDENT PHOSPHOLIPASE A2b-KNOCKOUT MICE Goichi Beck 1, Koei Shinzawa 2, Hisae Sumi-Akamaru 3, Hideki Mochizuki 3. 1 Department of Neurology, Toyonaka Municipal Hospital, Toyonaka, Japan; 2 Department of Medical Genetics, Osaka University Graduate School of Medicine, Suita, Japan; 3 Department of Neurology, Osaka University Graduate School of Medicine, Suita, Japan
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Objectives: It has been reported that mutations in the PLA2G6 gene for calcium-independent phospholipase A2b (iPLA2b) are responsible for autosomal recessive L-dopa responsive parkinsonism [1], suggesting the presence of some neurodegenerative conditions in the striatonigral dopaminergic system in patients with PLA2G6 mutations. The objective of this study is to clarify whether deficiency of iPLA2b would lead to degeneration of the striatonigral dopaminergic neurons. Methods: We performed an immunohistochemical analysis in the striatum and substantia nigra (SN) of iPLA2b-knockout (KO) mice. Results: In iPLA2b-KO mice, focal loss of nerve terminals positive for tyrosine hydroxylase (TH) and dopamine transporter (DAT) was found from 56 weeks (early clinical stage), although iPLA2b-KO mice demonstrated no significant reduction in the number of dopaminergic neurons in SN in comparison with age-matched wild-type mice. At 100 weeks (late clinical stage), more severe reductions in the level of immunoreactivities for DAT were observed compared with that for TH in the striatum of iPLA2b-KO mice. Moreover, strongly TH-positive structures that were supposed to be deformed axons were observed in the neuropil of the striatum of KO mice from 15 weeks (the preclinical stage) that increased with age. Conclusions: These results suggest that progressive degeneration of dopaminergic neurons would mainly occur in the distal part of axons in iPLA2b-KO mice. High TH expression and downregulation of DAT in the deformed axons may be a compensatory mechanism. References: 1. Paisan-Ruiz C, et al. Ann Neurol 2009. P 6.016. C-REL DEFICIENT MICE, A MOUSE MODEL OF “SPREADING” PD-LIKE PATHOLOGY Edoardo Parrella 1, Arianna Bellucci 1, Vanessa Porrini 2, Annamaria Lanzillotta 1, Marina Benarese 1, Gaia Vegezzi 1, Cristina Baiguera 1, Angelo Antonini 3, Pier Franco Spano 2, Marina Pizzi 2. 1 Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy; 2 Department of Molecular and Translational Medicine, University of Brescia; IRCCS, Ospedale San Camillo, Brescia Venezia, Italy; 3 IRCCS Ospedale San Camillo, Venezia, Italy Objectives: At premotor pathological stages, many PD patients suffer from nonmotor manifestations, including olfactory dysfunction and gastrointestinal constipation (1). In many PD cases synucleinopathy begins in defined nervous sites and progress in an anatomically predictable sequence, spreading from periphery (olfactory structures and enteric nervous system) to superior brain regions (2). NF-kB factors are cardinal players in the progression of the neurodegenerative process. In particular, the c-Rel subunit plays a crucial neuroprotective role, contributing to brain resilience to stress. We have previously shown that mice lacking c-Rel (crel-/- mice) develop with aging DA neuronal loss in substantia nigra pars compacta (SNc) with accumulation of aggregated alpha-synuclein, microglia activation and motor deficits responsive to L-DOPA administration (3). Methods: WT and c-rel-/- mice were studied at different ages with behavioral tests and biochemical /immunohistochemistry techniques. Moreover, levels and activity of c-Rel were analyzed in blood cells and brain samples of PD patients. Results: Already at a premotor pathology stage, c-rel-/- mice displayed olfactory and gastrointestinal dysfunctions, accumulation of alpha-synuclein in olfactory bulb as well as in the dorsal nucleus of vagus and locus coeruleus, striatal loss of dopamine transporter. Furthermore, the activity of the c-Rel protein decreased in the blood cells and SN of a considerable number of PD patients included in the study. Conclusions: c-rel-/- mice represent an innovative animal model to study pathological progression of PD and to investigate novel therapeutic approaches for early intervention in this pathology. Our recent data on humans suggest that c-Rel deficiency might be relevant also in the pathology of PD patients. References: 1. Simuni and Sethi. Ann Neurol 2008. 2. Braak et al. Cell Tissue Res 2004. 3. Baiguera et al. Brain 2012
Objectives: In accordance with modern basal ganglia model, genetic code alteration responsible for formation of corticobasal circuit should lead to changes of movement and behavior. Methods: Within 3 years, line of outbred mice, which were inbreed for 8 generations (16 mutant mice and 10 controls) was studied. Weight monitoring and open field test were done. Assessment of social behavior was analyzed by numbers of newborn mice. Quality of mutation was determined by direct and backcrossing in 3 generations. Results: Selection showed that mutation is spontaneous and recessive linked to X chromosome. Open field test showed that 2 out of 16 mutants were able to get out (6.3 min, 1.4 min control). Control group showed normal social behavior, number of mice born was 7 in average, average generation time was 78 days. Mutants have 6 mice, but as females didn't express parental instinct, number of two week age mice reached 0. All mutants were grown up by either control mice or carriers of mutant gene. Generation time e 147d. Weight monitoring isn't informative. Phenotype is formed by 4th week of development with progresses to the second month, manifested by stereotypical circular movements in both directions, and inability to walk in straight line, the diameter of the circle depends on the degree of animals' anxiety. Calm animal perform pendulum motion with linear trajectory. Frightened animal rotates around gravity axis. Conclusions: Considering the phenotype of mutation, we assumed a defect in the structure of reelin. This mutation would be good model for stereotyping movement disorders. P 6.014. GENETIC MODELS OF PARKINSON'S DISEASE Moein Dehbashi 1, 3, Maedeh Rouigari 2,3. 1 Genetics Division, Biology Department, Faculty of Science, University of Isfahan, Isfahan, Iran; 2 Biology Department, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran; 3 Isfahan Neurosciences Research Center, Al-Zahra Hospital, Isfahan University of Medical Sciences, Isfahan, Iran Objectives: The disappointing outcomes obtained so far by modelling the genetics of Parkinson's disease (PD) in mice have motivated interest in developing alternative genetic models in multicellular model organisms. Methods: Here we will focus on the fruit fly (Drosophila melanogaster), the nematode Caenorhabditis Elegans and the zebrafish (Danio Rerio) as the PD models. Results: The fruit fly (D. melanogaster), the nematode C. Elegans and the zebrafish (Danio Rerio) exhibit some clear advantages over rodents in terms of the relative ease with which the genome can be manipulated and of the much reduced costs involved in the development of genetic models of PD, but of course, their face validity is limited by the nature of the ‘symptoms’ these species present with. Conclusions: Given that these models are in much earlier stages of development, they have yet to play a role in drug discovery for PD; however, they may prove invaluable in the future development of diseasemodifying strategies that have so far yielded little success in clinical efforts. References: 1. Dawson TM, Ko HS, Dawson VL. Genetic animal models of Parkinson's disease. Neuron 2010;66:646-661. 2. Duty S, Jenner P. Animal models of Parkinson's disease: a source of novel treatments and clues to the cause of the disease. Br J Pharmacol 2011;164:1357-1391. 3. Le W, Sayana P, Jankovic J. Animal models of Parkinson's disease: a gateway to therapeutics? Neurotherapeutics 2014;11:92-110. P 6.015. PROGRESSIVE AXONAL DEGENERATION OF THE STRIATONIGRAL DOPAMINERGIC NEURONS IN CALCIUM-INDEPENDENT PHOSPHOLIPASE A2b-KNOCKOUT MICE Goichi Beck 1, Koei Shinzawa 2, Hisae Sumi-Akamaru 3, Hideki Mochizuki 3. 1 Department of Neurology, Toyonaka Municipal Hospital, Toyonaka, Japan; 2 Department of Medical Genetics, Osaka University Graduate School of Medicine, Suita, Japan; 3 Department of Neurology, Osaka University Graduate School of Medicine, Suita, Japan
e179
Objectives: It has been reported that mutations in the PLA2G6 gene for calcium-independent phospholipase A2b (iPLA2b) are responsible for autosomal recessive L-dopa responsive parkinsonism [1], suggesting the presence of some neurodegenerative conditions in the striatonigral dopaminergic system in patients with PLA2G6 mutations. The objective of this study is to clarify whether deficiency of iPLA2b would lead to degeneration of the striatonigral dopaminergic neurons. Methods: We performed an immunohistochemical analysis in the striatum and substantia nigra (SN) of iPLA2b-knockout (KO) mice. Results: In iPLA2b-KO mice, focal loss of nerve terminals positive for tyrosine hydroxylase (TH) and dopamine transporter (DAT) was found from 56 weeks (early clinical stage), although iPLA2b-KO mice demonstrated no significant reduction in the number of dopaminergic neurons in SN in comparison with age-matched wild-type mice. At 100 weeks (late clinical stage), more severe reductions in the level of immunoreactivities for DAT were observed compared with that for TH in the striatum of iPLA2b-KO mice. Moreover, strongly TH-positive structures that were supposed to be deformed axons were observed in the neuropil of the striatum of KO mice from 15 weeks (the preclinical stage) that increased with age. Conclusions: These results suggest that progressive degeneration of dopaminergic neurons would mainly occur in the distal part of axons in iPLA2b-KO mice. High TH expression and downregulation of DAT in the deformed axons may be a compensatory mechanism. References: 1. Paisan-Ruiz C, et al. Ann Neurol 2009. P 6.016. C-REL DEFICIENT MICE, A MOUSE MODEL OF “SPREADING” PD-LIKE PATHOLOGY Edoardo Parrella 1, Arianna Bellucci 1, Vanessa Porrini 2, Annamaria Lanzillotta 1, Marina Benarese 1, Gaia Vegezzi 1, Cristina Baiguera 1, Angelo Antonini 3, Pier Franco Spano 2, Marina Pizzi 2. 1 Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy; 2 Department of Molecular and Translational Medicine, University of Brescia; IRCCS, Ospedale San Camillo, Brescia Venezia, Italy; 3 IRCCS Ospedale San Camillo, Venezia, Italy Objectives: At premotor pathological stages, many PD patients suffer from nonmotor manifestations, including olfactory dysfunction and gastrointestinal constipation (1). In many PD cases synucleinopathy begins in defined nervous sites and progress in an anatomically predictable sequence, spreading from periphery (olfactory structures and enteric nervous system) to superior brain regions (2). NF-kB factors are cardinal players in the progression of the neurodegenerative process. In particular, the c-Rel subunit plays a crucial neuroprotective role, contributing to brain resilience to stress. We have previously shown that mice lacking c-Rel (crel-/- mice) develop with aging DA neuronal loss in substantia nigra pars compacta (SNc) with accumulation of aggregated alpha-synuclein, microglia activation and motor deficits responsive to L-DOPA administration (3). Methods: WT and c-rel-/- mice were studied at different ages with behavioral tests and biochemical /immunohistochemistry techniques. Moreover, levels and activity of c-Rel were analyzed in blood cells and brain samples of PD patients. Results: Already at a premotor pathology stage, c-rel-/- mice displayed olfactory and gastrointestinal dysfunctions, accumulation of alpha-synuclein in olfactory bulb as well as in the dorsal nucleus of vagus and locus coeruleus, striatal loss of dopamine transporter. Furthermore, the activity of the c-Rel protein decreased in the blood cells and SN of a considerable number of PD patients included in the study. Conclusions: c-rel-/- mice represent an innovative animal model to study pathological progression of PD and to investigate novel therapeutic approaches for early intervention in this pathology. Our recent data on humans suggest that c-Rel deficiency might be relevant also in the pathology of PD patients. References: 1. Simuni and Sethi. Ann Neurol 2008. 2. Braak et al. Cell Tissue Res 2004. 3. Baiguera et al. Brain 2012