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Oxidation of DJ-1-dependent regulation of tyrosine hydroxylase
S246
Abstracts
the role of ␣-syn in the axonal pathology, Thy-1 promoter driven ␣-syn transgenic (Tg) mice were analyzed by histological procedures. Two types of ␣-syn accumulated abnormal axon structures, possibly synaptic swellings, were observed in the brains of 18 months old Tg mice but not in the control mice. One was with about 2 m diameter structure, which was immunopositive for vesicular glutamate transporter. The other had 4 m or more diameter and was immunostained with glutamic-acid decarboxylase. Interestingly, calbindin was positive only in the larger type of swelling. Further ultrastructural characterization of these two types of ␣-syn accumulated terminals is in progress. Our results suggest that at least two types of axonal pathology may be brought about by ␣-syn accumulation in the Tg mice brain. doi:10.1016/j.neures.2009.09.1392
P3-l17 Analysis of the degradation of ␣-synuclein by neurosin Harutsugu Tatebe 1 , Yoshihisa Watanabe 2 , Takashi Kasai 1 , Takahiko Tokuda 1 , Toshiki Mizuno 1 , Masaki Tanaka 2 , Masanori Nakagawa 1 1
Dept. Neurol., Kyoto Pref. Univ. Med., Japan; Geriatr., Kyoto Pref. Univ. Med., Japan
2
Dept. Cell Biol., Res. Inst.
␣-Synuclein (␣-syn) is known to be the major component of Lewy Body, which is commonly observed in dopaminergic neurons of patients in sporadic Parkinson’s disease (PD). Usually abnormal ␣-syn is degraded by certain catabolic pathways. In the present study, we studied the degradation of ␣-syn by neurosin (NS), a member of the kallikrein family of serine proteases, and revealed that NS cleaved within the NAC region of ␣-syn in vitro. We then investigated the localization and proteolytic activity of NS using HEK293T cells. The exogenously expressed NS was predominantly secreted from cells. A fraction of NS was localized to the ER and the Golgi apparatus. Proteolytic activities of intra- and extracellular NS were analyzed by zymography, resulting that extracellular NS had protease activity but intracellular NS did not. This result suggests that NS targets to the extracellular ␣-syn. doi:10.1016/j.neures.2009.09.1393
hyper-oxidized form of DJ-1, namely at C106, binds to TH but not stimulates the activity. doi:10.1016/j.neures.2009.09.1395
P3-l20 Experimental analysis of anti-parkinsonian effects of zonisamide Hideto Miwa, Tomomi Kubo, Kiwa Kobayashi, Yoshinori Kajimoto, Tomoyoshi Kondo Dept. Neurol., Wakayama Med. Univ., Wakayama, Japan Objectives: Zonisamide (ZNS), an anti-epileptic drug, has beneficial efficacy in Parkinson disease. Its pharmacological mechanisms remain unclear. The present study aims to study the effect of ZNS on experimental tremors. Methods: Effect of ZNS on harmaline-induced tremor and tacrine-induced tremulous jaw movements (TJMs) in rats, an experimental model of parkinosnian tremor, was studied. Results: ZNS significantly suppressed both harmaline-induced tremor and tacrineinduced TJMs, and this effect was not lost under conditions of monoamine-depletion. There was no effect of ZNS on harmaline-induced and tacrine-induced c-Fos expression in the primary site of the pharmacological action, such as the olivary nucleus and ventrolateral striatum, respectively. ZNS suppressed the tacrine-induced c-Fos expression in the cortex, the dorsal striatum, and the nucleus accumbens, which are involved in the architecture of the cortico-basal ganglia-thalamocortical circuits. Conclusion: ZNS has the anti-tremor effect that may be due to a broad inhibitory effect on tremor-related structures. doi:10.1016/j.neures.2009.09.1396
P3-m01 STN-DBS affects the REM switch Namiko Nishida 1 , Hiroki Toda 1 , Hidemoto Saiki 2 , Tokiko Murakami 3 , Kunihiro Kadoh 3 , Hideki Hayashi 1 , Keita Ueda 4 , Sadayuki Matsumoto 2 , Jun Takahashi 1 1
P3-l18 Prevention of MPTP-induced parkinsonism by calbindin recruitment into nigral dopaminergic cells Kenichi Inoue 1,2 , Shigehiro Miyachi 2,3 , Katsunori Nishi 1 , Haruo Okado 1 , Atsushi Nambu 4 , Masahiko Takada 1,2 1
Dept. Syst. Neurosci., Tokyo Met. Inst. Neurosci., Tokyo, Japan; 2 CREST, JST, Japan; 3 Dept. Behav. Brain Sci., Pri. Res. Inst., Kyoto Univ., Kyoto, Japan; 4 Div. Syst. Neurophysiol, NIPS, Okazaki, Japan A specific population of dopaminergic cells in the substantia nigra pars compacta that expresses calbindin selectively survives during the cell death period in Parkinson’s disease. Based on this finding, we examined the preventive effect of calbindin (Cb) recruitment into nigral dopaminergic cells, with an adenoviral vector and an lentiviral vector, on parkinsonian insults induced by 1-methyl-4-phenyl-1,2,3,6tetrahydropyridine (MPTP). Our behavioral analysis showed that a series of motor impairments occurred primarily in the limbs ipsilateral to Cb recruitment. Furthermore, our immunohistochemical analysis revealed that the expression level of tyrosine hydroxylase in the nigrostriatal system was preserved to some extent on the Cb-recruited side. The present results indicate that Cb recruitment into nigral dopaminergic cells with recombinant viral vectors may provide an effective therapeutic approach to Parkinson’s disease. doi:10.1016/j.neures.2009.09.1394
P3-l19 Oxidation of DJ-1-dependent regulation of tyrosine hydroxylase Shizuma Ishikawa 1 , Takahiro Taira 2 , Hiroyoshi Ariga 1 , Sanae M.M. Iguchi-Ariga 3 1
Grad. Pharm. Sci. Hokkaido Univ., Japan; Japan; 3 Grad. Agr. Hokkaido. Univ., Japan
2
Grad. Med. Yamanashi. Univ.,
DJ-1 is a multi-functional protein and plays roles in transcriptional regulation and anti-oxidative stress, loss of which is thought to be the onset of Parkinson’s disease. Although tyrosine hydroxylase (TH) is a key enzyme for dopamine biosynthesis, it is not clarified yet whether DJ-1 regulates TH. We therefore examined the effect of DJ-1 on TH activity. When SH-SY5Y cells were exposed to H2 O2 at various concentrations, TH activity increased to the maximal at 10 M H2 O2 , and then decreased to the level lower than that without H2 O2 -treatment. The complex formation between DJ-1 and TH, on the other hand, was increased with the increased amount of H2 O2 in a dose-dependent manner. These results suggest that the degree of DJ-1 oxidation, rather than the amount of the DJ-1–TH complex, affects TH activity: the
Dept. Neurosurg., Kitano Hospital, Osaka, Japan; 2 Dept. Neurol, Kitano Hospital, Japan; 3 Dept. Lab. Med., Kitano Hospital, Japan; 4 Dept. Psychiatry, Kitano Hospital, Japan Sleep disturbances are common in Parkinson’s disease(PD). Neuronal degeneration in PD progresses from caudal brainstem possibly including caudal REM-on and rostral REM-off centers. The aim of this study was to determine the effect of deep brain stimulation of the subthalamic nucleus (STN-DBS) on sleep symptoms. We assessed subjective symptoms with PD sleep scale(PDSS) on 17 patients, and objective symptoms on 7 patients with polysomnography (PSG) before and after surgery. Medications and motor symptoms were assessed by levodopa equivalent daily dose (LEDD) and by Unified PD Rating Scale (UPDRS). PDSS correlated with UPDRS motor part, and improved significantly. PSG showed improved sleep architecture with increase in the REM percentage, which was inversely correlated with PD duration. This study indicates that not only the alleviated motor symptoms but also the viable REM-on neurons could have contributed to the sleep modification. doi:10.1016/j.neures.2009.09.1397
P3-m02 Accumulation of C-terminal ATN1 fragment in DRPLA Yasuyo Suzuki 1 , Kimiko Nakayama 1 , Naohiro Hashimoto 2 , Ikuru Yazawa 1 1 Lab. of Research Resources, Natl. Center for Geriatrics and Gerontology, Obu, Japan; 2 Dep. of Regenerative Med., Natl. Center for Geriatrics and Gerontology, Obu, Japan
Dentatorubral-pallidoluysian atrophy (DRPLA) is a hereditary neurodegenerative disorder. The responsible gene product (DRPLA protein), also known as atrophin1 (ATN1), carry polyglutamine (polyQ) expansion. Nuclear accumulation of ATN1 might be a primary disease process in DRPLA, but the mechanism of neurodegeneration is still unclear. Using COS-7 and Neuro2a cells expressing the ATN1 gene, we demonstrated that ATN1 was accumulated with increasing number of polyQs, and identified a novel C-terminal fragment containing a polyQ tract. The mutant C-terminal fragment with expanded polyQ selectively accumulated in the cells, and it was showed in the brain tissues of patients with DRPLA. Full-length ATN1 and C-terminal fragment displayed individual localization, and the mutant fragment was preferentially found in the membrane fraction. We suggest that the mutant fragment plays a role in DRPLA neurodegeneration. doi:10.1016/j.neures.2009.09.1398
Abstracts
the role of ␣-syn in the axonal pathology, Thy-1 promoter driven ␣-syn transgenic (Tg) mice were analyzed by histological procedures. Two types of ␣-syn accumulated abnormal axon structures, possibly synaptic swellings, were observed in the brains of 18 months old Tg mice but not in the control mice. One was with about 2 m diameter structure, which was immunopositive for vesicular glutamate transporter. The other had 4 m or more diameter and was immunostained with glutamic-acid decarboxylase. Interestingly, calbindin was positive only in the larger type of swelling. Further ultrastructural characterization of these two types of ␣-syn accumulated terminals is in progress. Our results suggest that at least two types of axonal pathology may be brought about by ␣-syn accumulation in the Tg mice brain. doi:10.1016/j.neures.2009.09.1392
P3-l17 Analysis of the degradation of ␣-synuclein by neurosin Harutsugu Tatebe 1 , Yoshihisa Watanabe 2 , Takashi Kasai 1 , Takahiko Tokuda 1 , Toshiki Mizuno 1 , Masaki Tanaka 2 , Masanori Nakagawa 1 1
Dept. Neurol., Kyoto Pref. Univ. Med., Japan; Geriatr., Kyoto Pref. Univ. Med., Japan
2
Dept. Cell Biol., Res. Inst.
␣-Synuclein (␣-syn) is known to be the major component of Lewy Body, which is commonly observed in dopaminergic neurons of patients in sporadic Parkinson’s disease (PD). Usually abnormal ␣-syn is degraded by certain catabolic pathways. In the present study, we studied the degradation of ␣-syn by neurosin (NS), a member of the kallikrein family of serine proteases, and revealed that NS cleaved within the NAC region of ␣-syn in vitro. We then investigated the localization and proteolytic activity of NS using HEK293T cells. The exogenously expressed NS was predominantly secreted from cells. A fraction of NS was localized to the ER and the Golgi apparatus. Proteolytic activities of intra- and extracellular NS were analyzed by zymography, resulting that extracellular NS had protease activity but intracellular NS did not. This result suggests that NS targets to the extracellular ␣-syn. doi:10.1016/j.neures.2009.09.1393
hyper-oxidized form of DJ-1, namely at C106, binds to TH but not stimulates the activity. doi:10.1016/j.neures.2009.09.1395
P3-l20 Experimental analysis of anti-parkinsonian effects of zonisamide Hideto Miwa, Tomomi Kubo, Kiwa Kobayashi, Yoshinori Kajimoto, Tomoyoshi Kondo Dept. Neurol., Wakayama Med. Univ., Wakayama, Japan Objectives: Zonisamide (ZNS), an anti-epileptic drug, has beneficial efficacy in Parkinson disease. Its pharmacological mechanisms remain unclear. The present study aims to study the effect of ZNS on experimental tremors. Methods: Effect of ZNS on harmaline-induced tremor and tacrine-induced tremulous jaw movements (TJMs) in rats, an experimental model of parkinosnian tremor, was studied. Results: ZNS significantly suppressed both harmaline-induced tremor and tacrineinduced TJMs, and this effect was not lost under conditions of monoamine-depletion. There was no effect of ZNS on harmaline-induced and tacrine-induced c-Fos expression in the primary site of the pharmacological action, such as the olivary nucleus and ventrolateral striatum, respectively. ZNS suppressed the tacrine-induced c-Fos expression in the cortex, the dorsal striatum, and the nucleus accumbens, which are involved in the architecture of the cortico-basal ganglia-thalamocortical circuits. Conclusion: ZNS has the anti-tremor effect that may be due to a broad inhibitory effect on tremor-related structures. doi:10.1016/j.neures.2009.09.1396
P3-m01 STN-DBS affects the REM switch Namiko Nishida 1 , Hiroki Toda 1 , Hidemoto Saiki 2 , Tokiko Murakami 3 , Kunihiro Kadoh 3 , Hideki Hayashi 1 , Keita Ueda 4 , Sadayuki Matsumoto 2 , Jun Takahashi 1 1
P3-l18 Prevention of MPTP-induced parkinsonism by calbindin recruitment into nigral dopaminergic cells Kenichi Inoue 1,2 , Shigehiro Miyachi 2,3 , Katsunori Nishi 1 , Haruo Okado 1 , Atsushi Nambu 4 , Masahiko Takada 1,2 1
Dept. Syst. Neurosci., Tokyo Met. Inst. Neurosci., Tokyo, Japan; 2 CREST, JST, Japan; 3 Dept. Behav. Brain Sci., Pri. Res. Inst., Kyoto Univ., Kyoto, Japan; 4 Div. Syst. Neurophysiol, NIPS, Okazaki, Japan A specific population of dopaminergic cells in the substantia nigra pars compacta that expresses calbindin selectively survives during the cell death period in Parkinson’s disease. Based on this finding, we examined the preventive effect of calbindin (Cb) recruitment into nigral dopaminergic cells, with an adenoviral vector and an lentiviral vector, on parkinsonian insults induced by 1-methyl-4-phenyl-1,2,3,6tetrahydropyridine (MPTP). Our behavioral analysis showed that a series of motor impairments occurred primarily in the limbs ipsilateral to Cb recruitment. Furthermore, our immunohistochemical analysis revealed that the expression level of tyrosine hydroxylase in the nigrostriatal system was preserved to some extent on the Cb-recruited side. The present results indicate that Cb recruitment into nigral dopaminergic cells with recombinant viral vectors may provide an effective therapeutic approach to Parkinson’s disease. doi:10.1016/j.neures.2009.09.1394
P3-l19 Oxidation of DJ-1-dependent regulation of tyrosine hydroxylase Shizuma Ishikawa 1 , Takahiro Taira 2 , Hiroyoshi Ariga 1 , Sanae M.M. Iguchi-Ariga 3 1
Grad. Pharm. Sci. Hokkaido Univ., Japan; Japan; 3 Grad. Agr. Hokkaido. Univ., Japan
2
Grad. Med. Yamanashi. Univ.,
DJ-1 is a multi-functional protein and plays roles in transcriptional regulation and anti-oxidative stress, loss of which is thought to be the onset of Parkinson’s disease. Although tyrosine hydroxylase (TH) is a key enzyme for dopamine biosynthesis, it is not clarified yet whether DJ-1 regulates TH. We therefore examined the effect of DJ-1 on TH activity. When SH-SY5Y cells were exposed to H2 O2 at various concentrations, TH activity increased to the maximal at 10 M H2 O2 , and then decreased to the level lower than that without H2 O2 -treatment. The complex formation between DJ-1 and TH, on the other hand, was increased with the increased amount of H2 O2 in a dose-dependent manner. These results suggest that the degree of DJ-1 oxidation, rather than the amount of the DJ-1–TH complex, affects TH activity: the
Dept. Neurosurg., Kitano Hospital, Osaka, Japan; 2 Dept. Neurol, Kitano Hospital, Japan; 3 Dept. Lab. Med., Kitano Hospital, Japan; 4 Dept. Psychiatry, Kitano Hospital, Japan Sleep disturbances are common in Parkinson’s disease(PD). Neuronal degeneration in PD progresses from caudal brainstem possibly including caudal REM-on and rostral REM-off centers. The aim of this study was to determine the effect of deep brain stimulation of the subthalamic nucleus (STN-DBS) on sleep symptoms. We assessed subjective symptoms with PD sleep scale(PDSS) on 17 patients, and objective symptoms on 7 patients with polysomnography (PSG) before and after surgery. Medications and motor symptoms were assessed by levodopa equivalent daily dose (LEDD) and by Unified PD Rating Scale (UPDRS). PDSS correlated with UPDRS motor part, and improved significantly. PSG showed improved sleep architecture with increase in the REM percentage, which was inversely correlated with PD duration. This study indicates that not only the alleviated motor symptoms but also the viable REM-on neurons could have contributed to the sleep modification. doi:10.1016/j.neures.2009.09.1397
P3-m02 Accumulation of C-terminal ATN1 fragment in DRPLA Yasuyo Suzuki 1 , Kimiko Nakayama 1 , Naohiro Hashimoto 2 , Ikuru Yazawa 1 1 Lab. of Research Resources, Natl. Center for Geriatrics and Gerontology, Obu, Japan; 2 Dep. of Regenerative Med., Natl. Center for Geriatrics and Gerontology, Obu, Japan
Dentatorubral-pallidoluysian atrophy (DRPLA) is a hereditary neurodegenerative disorder. The responsible gene product (DRPLA protein), also known as atrophin1 (ATN1), carry polyglutamine (polyQ) expansion. Nuclear accumulation of ATN1 might be a primary disease process in DRPLA, but the mechanism of neurodegeneration is still unclear. Using COS-7 and Neuro2a cells expressing the ATN1 gene, we demonstrated that ATN1 was accumulated with increasing number of polyQs, and identified a novel C-terminal fragment containing a polyQ tract. The mutant C-terminal fragment with expanded polyQ selectively accumulated in the cells, and it was showed in the brain tissues of patients with DRPLA. Full-length ATN1 and C-terminal fragment displayed individual localization, and the mutant fragment was preferentially found in the membrane fraction. We suggest that the mutant fragment plays a role in DRPLA neurodegeneration. doi:10.1016/j.neures.2009.09.1398