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Grtmifibrillary acidic protein (GFAP) and Heat shock protein 70 (HSP70) was studiedusing immunocytofluorescence. After exposure of the C6 glioma cells to glutamate

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3-m16 Involvement of KLK6 in the pathogenesis of experimentalutoimmune encephalomyelitisoshio Bando, Koichi Murakami, Shigetaka Yoshida

Dept. Functional Anatomy and Neuroscience, Asahikawa Medical College,sahikawa, Japan

ultiple Sclerosis (MS) is an inflammatory demyelinating disease of the centralervous system (CNS). However, molecular mechanism of demyelination has noteen well understood. Kalikrein 6 (KLK6) is a kalikrein-like serine protease that isistributed exclusively in oligodendrocytes in the CNS. To investigate the role ofLK6 in demyelination, we examined the expression of KLK6 in myelin oligoden-rocytes glycoprotein (MOG) induced experimental autoimmune encephalomyelitisEAE). In wild-type mice, KLK6 is induced in the spinal cord of mice with EAE. KLK6nock out (KO) mice exhibited an altered EAE progression characterized by delayednset and progression of clinical symptoms as compared to wild-type mice. Thesebservations suggest that KLK6 is involved in the pathogenesis of EAE mediated byemyelination.

oi:10.1016/j.neures.2009.09.1412

3-m17 The lineage tracing of Olig2+ cells in a murine demyelinatingisease modelakahiro Shimizu1,2, Kenji Tanaka1,2, Hirohide Takebayashi3,azuhiro Ikenaka1,2

National Institute for Physiological Sciences, Aichi, Japan; 2 The Graduateniversity for Advanced Studies, Kanagawa, Japan; 3 Dept. of Morphologicaleural. Science, Kumamoto Univ., Kumamoto, Japan

ligodendrocytes are myelin forming cells in the CNS. Demyelinating lesions areormed after degeneration of myelin membrane. In chronic demyelinating lesions,ifferentiation and remyelination of oligodendrocytes are inhibited in spite of theresence of abundant oligodendrocyte progenitor cells (OPCs). We hypothesizedhat cell fate of OPCs had been changed because many reactive astrocytes werebserved in demyelinating lesions. To validate this hypothesis, we performed lin-age tracing of OPCs using the mouse in which CreER is knocked into the olig2ocus. Olig2 gene is essential for development and differentiation of oligodendro-ytes and is expressed in adult OPCs. We used plp1 over expressing mice for analysisf demyelinating/-ed brain. We concluded that inhibition of remyelination is causedy the degeneration of immature or mature oligodendrocyte, not by the failure ofligodendrocyte supply due to the cell fate alteration of OPCs into astrocytes.

oi:10.1016/j.neures.2009.09.1413

3-m18 Unique gene expression changes in microglia coincide withhe appearance of chronic demyelinating lesionazuhiro Ikenaka1,2, Jianmei Ma1, Takahiro Shimizu1,2, Kenjianaka1,2

National Institute for Physiological Sciences, Okazaki, Japan; 2 The Gradu-te University for Advances Studies, Okazaki, Japan

ystatin F is papain-like lysosomal cysteine proteinase inhibitors. We found expres-ion of Cystatin F was totally absent in the normal brain and its induction in microgliaas dependent on phagocytosis of compact myelin membrane in either plptg/− micer cuprizone induced demyelinating model mice. Its expression was mostly absent

n brains of dysmyelinating jimpy and shiverer mutant mice, where there is veryittle compact myelin membrane even though microglia are activated. This stronglyuggests that microglia can be activated in a specific manner when they phagocytoseegenerated compact myelin membrane. At later stage of demyelination in plptg/−ice and cuprizone fed mice, the level of cystatin F expression went down when

he remyelinating ability was lost. This demonstrates that the gene expression ofctivated microglia is not constant in vivo during the course of demyelination andhis change coincides with the appearance of chronic demyelinating lesions.

oi:10.1016/j.neures.2009.09.1414

3-m19 GM-CSF-stimulated microglia differentiate into dendritic-likentigen-presenting cellsua Li, Yoshifumi Sonobe, Hideyuki Takeuchi, Tetsuya Mizuno, Akiouzumura

Dept. Neuroimmunol., RIEM, Nagoya Univ., Japan

im: We examined whether microglia express dendritic cells (DCs) markers andunction as DC in the presence of GM-CSF.

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ts S249

ethods: Bone marrow-derived DC (BM-DC) and microglia were derived from57BL/6J mice. We assessed the mRNA expressions of DC markers on BM-DC andicroglia with RT-PCR using each specific primer of CD11c, Dec205, MHC-II, CD80,D86, or GAPDH. Microglia were assessed with a flowcytometer using antibodiespecific for mouse CD11c, CD11b, CD80, CD86, CD83, CD209, MHC-II.esults: GM-CSF-stimulated microglia express DC markers. They can proliferate Tells more effectively than non-stimulated microglia. Discussion: A variety of stimulieportedly induce GM-CSF release from CNS cells. Our data suggest that GM-CSF-timulated microglia differentiate into DC-like cells and function as APC in theathological conditions.onclusion: GM-CSF-stimulated microglia differentiate into DC-like cells and func-

ion as APC more effectively than non-stimulated microglia.

oi:10.1016/j.neures.2009.09.1415

3-m20 Humanin rescues cortical neurons from NMDA-mediatedxcitotoxicity by the MAPK pathwayiaorong Yang, Pengjun Zhang, Ye Wang, Ruihong Shi, Huapingin, Ce Zhang

Dept. of Neurobiology, Shanxi Medical University, Taiyuan, Shanxi, PR China

umanin (HN) is a novel survival protein that protects against cell death inducedy various Alzheimer’s disease (AD)-related insults. It has been proposed that HNinds to a putative receptor on the cell membrane and triggers a signal transduc-ion cascade linked to neuroprotection. The present study is to investigate whetherhe neuroprotective effects of HN is mediated by mitogen-activated protein kinasesMAPK) pathways by using an in vitro N-methyl-d-aspartate (NMDA)-mediated exci-otoxicity of cortical neurons model. Our results reveal that neuroprotection byN in NMDA-mediated excitotoxicity of cortical neurons is mediated by JNK and38 MAPK, while extracellular signal-regulated kinase (ERK) did not involve inN-mediated rescue of neuronal cell death induced by NMDA-related insults. Under-

tanding the function of HN and its downstream pathway should lead to effectiventi-neurodegenerative diseases therapeutics.

oi:10.1016/j.neures.2009.09.1416

3-n01 1,2-dichloroethane induced toxic encephalopathyiao Niu1, Qinli Zhang1, Laiyu Li2, Lijun Yang1, Xiaoli Guo1,ouxin Liang3, Jianxun Huang2

Department of Occupational health, Shanxi Medical University, Taiyuan,hina; 2 Guangdong Provincial Institute of Occupational Health, China;Fudan University, China

he overall objective of this investigation was to characterize the acute neurotoxic-ty of 1,2-dichloroethane inhalation. Assessment of possible clinical and pathologicalffects on 1,2-DCE exposed workers was performed. 1,2-DCE poisoned model wasade in rats. Autopsy on a 1,2-DCE induced victim showed that obvious edemaas found. Sprague-Dawley rat animal model of 1,2-DCE cerebral edema was dupli-

ated with static inhalation. Pathological examination indicated that there wererain lesions such as edema, necrosis and hemorrhage. Microscopic photographsemonstrated that gaps between cells and vessels were obviously widened. Elec-ronic microscope observation indicated that nerve fibers were demyelinated andhe axis cylinder was pushed sideward. Lysis of cytoplasm, swelling mitochondriand obvious derangement of mitochondrial cristae were observed and cavities ofough surfaced endoplasmic reticulum were enlarged.

oi:10.1016/j.neures.2009.09.1417

3-n02 Ashwagandha extract protects against glutamate-inducedytotoxicity in retinoic acid differentiated C6 glioma cellsardeep Kataria, Gurcharan Kaur

Guru Nanak Dev University, India

lutamate neurotoxicity has been implicated in stroke, head trauma, multiple scle-osis and neurodegenerative disorders. The cytoprotective effects against glutamateoxicity were studied using the extract from Ashwagandha, a traditional Indianedicinal herb in the retinoic acid differentiated C6 glioma cells. The cytotoxic-

ty was evaluated using MTT and LDH assays. Furthermore the expression of glial

100 �M–10 mM), cell death was observed as evidenced by alteration of cell morphol-gy and nuclear condensation with Hoechst staining. Pre-treatment of the culturesith Ashwagandha extract (0.05 and 0.1%) significantly elevated the cell viability.