e306 Abstracts / Neuroscience Research 68S (2010) e223–e334 species-specific transcriptional regulation of the TH promoter by DJ-1 and one of mechanism for no reduction of TH in DJ-1 knockout mice. doi:10.1016/j.neures.2010.07.1356 P2-n18 Endogenous expression of neurotrophic factors in reactive astrocytes in 6-hydroxydopamine-lesioned rats Cathy N.P. Lui 1 , Tyler C.H. Leung 1 , L.W. Chen 2 , Ken K.L. Yung 1 1 Department of Biology, Hong Kong Baptist University 2 Institute of Neuro- sciences, The Forth Military Medical University, Xian, PR China PD is caused by a progressive loss of dopaminergic neurons in the substan- tia nigra (SN). At the early stages after lesion in the 6-hydroxydopamine (6-OHDA)-lesioned rat, nestin was found to re-express in the astrocytes of striatum (Str) and SN. These nestin-immunoreactive reactive astrocytes co-expressed brain-derived neurotrophic factor (BDNF) and glial cell-line derived neurotrophic factor (GDNF). Expressions of nestin in both Str and SN were found to be time-dependent. It transiently peaked at day 3 post lesion and diminished after day 5. Interestingly, BDNF and GDNF were started to express in the striatum and also in the substantia nigra during the peak expression of nestin-immunoreactive astrocytes. In the results of the west- ern blotting, expressions of mature forms of BDNF and GNDF were then found to increase progressively starting at day 3 and peak at day 7 post lesion. The present results reveal that nestin-positive reactive astrocytes are likely one of the sources of mature forms of BDNF and GDNF and may provide endoge- nous neurotrophic effects and self repair on axonal terminals and perikarya of dopaminergic neurons during very early stage of PD. doi:10.1016/j.neures.2010.07.1357 P2-n19 Generation of disease-specific induced pluripotent stem cells from hereditary Parkinson’s disease Takayuki Kondo 1,2 , Haruhisa Inoue 2,3 , Naohiro Egawa 1,2 , Kat- suhiro Yoshikawa 1 , Satoko Yamawaki 1 , Motoko Naitoh 1 , Shige- hiko Suzuki 1 , Kazutoshi Takahashi 2,3 , Kazuko Hasegawa 4 , Tatsu- toshi Nakahata 2 , Shinya Yamanaka 1 , Ryosuke Takahashi 2 1 Department of Neurology, Kyoto University School of Medicine 2 Center for iPS Cell Research and Application, Institute for Integrated Cell-Material Sciences, Kyoto University 3 JST-CREST 4 National Hospital Organization, Sagamaihara National Hospital Purpose: In Parkinson’s disease (PD) selective degeneration of midbrain dopaminergic neurons occur. However disease mechanism has not been clear yet. Disease-specific induced pluripotent stem cells (iPSCs) from hereditary PD patients will provide a great oppotiunity for explor- ing PD pathomechanism. A single amino acid mutation (I2020T) of Leucine-Rich Repeat Kinase 2 gene (LRRK2) is associated with heredi- tary PD in Kanagawa prefecture Japan. We establish iPSCS, derived from LRRK2-mutated PD patients, and analyze LRRK2 physiological function and mechanism of mutant LRRK2-related midbrain dopaminergic neuron degeneration. Methods and results: We cultivated skin fibroblast, obtained from Sagamihara-hereditary PD member (s). Next we transfected retrovirus vec- tors (carrying four transcriptional factors, including Oct3/4, SOX2, KLF4, and/or c-Myc) into fibroblast and seeded onto SNL feeder cells. Finally we isolate colonies, which have similar appearance to embryonic stem cells, and characterize isolated colonies. Conclusion: We established iPSCs, derived from PD patient(s) with I2020T mutation in LRRK2 gene. Disease-specific iPS cells from hereditary PD patient(s) may contribute to unraveling the pathogenesis of PD. doi:10.1016/j.neures.2010.07.1358 P2-n20 Polyunsaturated fatty acids induce oxidative stress and cell death in dopaminergic SH-SY5Y cells Masayo Shamoto-Nagai 1,2 , Yumi Kurokawa-Nose 1 , Makoto Naoi 3 , Wakako Maruyama 1 1 Dept Brain Sci, Natl Center Geriatr Gerontol, Aichi 2 JSPS Reseach Fellow, Tokyo 3 Dept Neurosci, Gifu Intern Inst Biotech, Gifu Selective degeneration of dopamine (DA) neurons and the accumulation of Lewy bodies (LB) in the substantia nigra are characteristic pathologi- cal features in Parkinson’s disease (PD). Oxidative stress, dysfunctions of mitochondria and ubquitine-proteasome system (UPS), and apoptotic sig- nal activation have been proposed as the pathogenesis of PD. Previously we show that rotenone, a mitochondria complex I inhibitor, induced dys- function of UPS through increased oxidative modification, accumulation and aggregation of proteins, and finally apoptotic cells death in SH-SY5Y cells. Oxidative stress increases cytotoxic lipid peroxides especially in the cellular membrane, which form adduct with proteins and change the conformation. Docosahexaenoic acid (DHA), a polyunsaturated fatty acid (PUFA), occurs abundantly in neuronal membrane. DHA is easily oxidized and its peroxida- tion product modifies lysine residue of proteins as porpanoyl-lysine (PRL). In this paper, we present the results on the effect of DHA to SH-SY5Y cells. The cells were incubated with various concentration of DHA in Cos-medium containing FBS and retinoic acid for 3 days. DHA increased cell death in a dose- dependent way, as assessed by LDH assay. The toxicity was suppressed by alpha-tocopherol and ascorbic acid. DHA at 10 M increased ATP synthesis, and decreased PRL-containing proteins in total cellular protein by ELISA sys- tem. On the other hand, high-doses of DHA decreased ATP level and increased PRL-proteins in mitochondrial membranous fraction. DHA reduced the activ- ity of 26S proteasome in SH-SY5Y cells transfected with a proteasome sensor vector encoding green fluorescence protein for a 26S proteasome substrate. These results suggest that PUFA may play a critical role in the degeneration of dopamine neurons in aging and PD by the oxidation in the mitochon- drial membrane, accumulation of the modified proteins, and the impaired function of ATP synthesis and UPS. doi:10.1016/j.neures.2010.07.1359 P2-n21 Induction of microsomal prostaglandin E synthase- 1 contributes to dopaminergic neuronal death in rats and mice mesencephalic neuronal culture treated with 6- hydroxydopamine Yuri Ikeda-Matsuo 1 , Tomoko Mizoguchi 1 , Satoshi Uematsu 2 , Shizuo Akira 2 , Yasuharu Sasaki 1 1 Lab. Pharmacol., Sch. Pharmac. Sci., Kitasato Univ., Tokyo, Japan 2 Lab. Host Defense, Immunol. Frontier Res. Center, Osaka Univ., Osaka, Japan We have recently demonstrated that microsomal prostaglandin E synthase-1 (mPGES-1) is a critical factor of stroke-reperfusion injury. In this study, we investigated the role of mPGES-1 in 6-hydroxydopamine (6-OHDA)-induced dopaminergic neuronal death in cultured mesencephalic neurons. The cul- tures from 14 to 15 embryonic mesencephalon were maintained for 7 days and then stimulated with 6-OHDA. Western blots of cultured rat mesen- cephalic neurons showed induction of mPGES-1 protein by 6-OHDA in a concentration-dependent manner. Immunostaining for mPGES-1 was also observed after 6-OHDA treatment. The mPGES-1-positive cells were also positive for MAP-2 and partially tyrosine hydroxylase (TH), marker proteins for neurons and dopamine neurons, respectively. 6-OHDA also increased both the activity of PGES and production of PGE 2 . The 6-OHDA-induced decrease of TH-positive cells was inhibited by NS-398, MK-886 and dex- amethasone, inhibitors for COX-2, mPGES-1 and induction of these proteins, respectively. These inhibitors also significantly inhibited 6-OHDA-induced PGE 2 production. These results suggest that 6-OHDA-induced dopaminergic neuronal death is mediated by induction of mPGES-1 through PGE 2 produc- tion. Therefore, we next examined the role of mPGES-1 in 6-OHDA-induced neurotoxicity using mPGES-1 knockout (KO) mesencephalic neurons. The production of PGE 2 observed in wild-type (WT) mesencephalic neurons was completely abolished in mPGES-1 KO neurons. Furthermore, 6-OHDA- induced dopaminergic neuronal death was significantly inhibited by deletion of mPGES-1. Treatment of PGE 2 with 6-OHDA increased neuronal death in mPGES-1 KO neurons to the same level of WT neurons, while PGE 2 itself had no effect on survival of neurons. Taken together, these results suggest that induction of mPGES-1 enhances dopaminergic neuronal death induced by 6-OHDA through PGE 2 production. Thus, mPEGS-1 may be a valuable therapeutic target for treatment of Parkinson’s disease. doi:10.1016/j.neures.2010.07.1360 P2-n22 Roles of neurokinin receptor agonists in pathogen- esis and therapy in models of Parkinsons disease John M.T. Chu , Ken K.L. Yung Depatment of Biology, Hong Kong Baptist University Parkinsons disease (PD) is a motor disorder and is caused by a degeneration of dopaminergic neurons in the substantia nigra pars compacta. Neurokinin (NK) is a group of neuropeptides including substance P (SP). Down-regulation of SP in the basal ganglia after onset of PD has been found and therefore NK1 receptor, the receptor for SP, is implicated in treatment of PD. The present study investigated the effects of septide, a selective NK1 agonist, in cell and