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Effects of D -serine on fear extinction in mice
S126
Abstracts
N-terminal peptide of mouse Prune2. Human PRUNE2 cDNA encodes a protein that consists of 3,063 amino acids. Western blotting demonstrated the presence endogenous Prune2 protein. Quantitative RT-PCR analysis revealed that the expression of PRUNE2 in brain is the highest among human tissues examined. We demonstrated that Prune2 is a huge protein with the Prune homology domain at the N-terminus and a BCL2/adenovirus E1B interacting protein 2 (BNIP2) and Cdc42GAP homology domain at the C-terminus. doi:10.1016/j.neures.2009.09.601
P1-m20 Jak signaling contributes to Concanavalin A induced C6 glioma cell death Kaoru Nagai, Hideki Yoshida, Yuta Inagawa, Takeo Kubota University of Yamanashi, Japan Because of the difficulty on the treatment of brain tumor including glioma, development of the methods to induce glioma cell death is highly desirable. We examined apoptotic effect of a mannose-binding plant lectin concanavalin A (ConA) on C6 glioma cells. ConA induced C6 cell death dose dependently via carbohydrate binding activity. DNA degradation and caspase-3 activation were observed in ConA treated cells. It indicates that ConA induced apoptotic cell death. Caspase-8 inhibitor significantly reduced the cell death, and a death receptor Fas in C6 cells was recognized by ConA. In addition, JAK inhibitor and gp130-Fc were also inhibited the cell death. Our data suggests that both death receptor and JAK signals contributes ConA induced glioma cell death, and ConA can be utilized to the glioma therapy. doi:10.1016/j.neures.2009.09.602
P1-n01 Interleukin-2 induced Jak-dependent cell death in C6 glioma cells via lectin activity Yuta Inagawa, Takeo Kubota, Kaoru Nagai University of Yamanashi, Japan Gliomas are the most common brain tumors. It was reported that glioma cells secrete molecules disrupting immune systems which contribute tumor elimination. Application of Interleukin-2 (IL-2) was reported to be effective on glioma treatment via stimulating antitumor immune response. However, the direct effect of IL-2 on glioma cells has not been elucidated, yet. In this study, we investigated the effects of IL-2 on C6 glioma cells. IL-2 reduced C6 cell number dose dependently. And the reduction was inhibited by JAK inhibitor, gp130-Fc, and caspase inhibitor. It suggests that IL-2 induced apoptotic cell death in C6 glioma cells via gp130-JAK signal. In addition, since IL-2 was reported to interact with cell surface high-mannose oligosaccharides, we evaluated if IL-2 functions via lectin activity. The haptenic sugar methyl-␣-mannoside significantly blocked the effect of IL-2. Our data suggests that therapeutic glioma treatment with IL-2 directly induced glioma cell death other than stimulating immune response. doi:10.1016/j.neures.2009.09.603
P1-n02 Effects of low-power laser irradiation on proliferation of human-derived glioblastoma A-172 Hideyuki Murayama 1 , Banri Yamanoha 2 , Kei Sadakane 2 , Shinichi Kogure 1 1 Department Bioinformatics, Graduate School of Engineering, Soka University, Tokyo, Japan; 2 Department Environmental Engineering for Symbiosis, Graduate School of Engineering, Soka University, Tokyo, Japan
It has been reported that low-power laser irradiation (LLI) can modulate various biological processes including cell proliferation. The present study was designed to examine effects of 808 nm (30 mW/cm2 ) LLI on the cell proliferation of humanderived glioblastoma A-172 in order to find a possibility for developing an LLI therapy for cancers. According to the conventional cell culture technique, A-172 cells with the same cell density were classified into four groups: control, group 1 (received 20 min dose of LLI), group 2 (40 min), and group 3 (60 min). Cell counting was performed on the PC screen and in some cases with MTT method. In the case of 808 nm LLI (n = 25), it decreased the proliferation ratio as compared with control group: A significant difference (p < 0.01) was obtained between all groups and control. It is concluded that LLI have a potential effect on the cell proliferation of human-derived glioblastoma A-172 with a dose-dependency. doi:10.1016/j.neures.2009.09.604
P1-n03 Electrolytic lesion of the dorsolateral periaqueductal gray attenuates the antinociceptive response of morphine microinjected into the nucleus cuneiformis Leila Ahmad-Molaei, Abbas Haghparast Neurosacience Research Center, Shahid Beheshti University, M.C., It has been indicated that periaqueductal gray (PAG) and nucleus cuneiformis (CnF) play well-established role in a network mediating anti- and pro-nociception. 32 adult male Wistar rats cannulated bilaterally into the CnF, concurrently lesion of dl-PAG was done. Formalin test was carried out to measure pain behavior and antinociceptive effect of morphine microinjected into the CnF (2.5 g/0.3 l saline per side). The results showed that dl-PAG lesion attenuated the effect of morphine microinjected into the CnF in formalin test while dl-PAG lesion solely did not alter basal pain behavior as compared with those in sham-lesion animals. In conclusion, our results suggest the existence of a direct or indirect projection from CnF to the dl-PAG at least at the level of the morphine antinociception in pain modulation. doi:10.1016/j.neures.2009.09.605
P1-n04 Effects of D-serine on fear extinction in mice Shingo Matsuda 1 , Nobuhisa Kanahara 2 , Daisuke Matsuzawa 1 , Ken Nakazawa 1 , Masaomi Iyo 2 , Eiji Shimizu 1 1
Department of Integrative Neurophysiology Graduate School of Medicine, Chiba University, Chiba, Japan; 2 Department of Psychiatry Graduate School of Medicine, Chiba University, Chiba, Japan Molecular mechanisms of fear extinction are not well known. To investigate effects of N-methyl-D-aspartate receptor (NMDA-R) partial agonist, D-serine, on fear extinction we examined cued fear conditioning and extinction paradigm in C57BL/6J mice and measured activation of cytosol and nuclear ERK 1/2 in hippocampus and amygdala. We found that administration of D-serine before extinction facilitated extinction of fear memory at recall tests. In both hippocampus and amygdala, Dserine significantly increased phosphorylation of ERK 1/2. The results suggested that D-serine may facilitate fear extinction through ERK signaling. doi:10.1016/j.neures.2009.09.606
P1-n05 Effects of Methomyl, agricultural chemicals, on learning in pigeons Tomoyuki Kanamatsu, Kentaro Oseki Department Environ. Engi. for Symbiosis, Faculty of Engi., Soka University, Tokyo, Japan The present study was designed to investigate the effects of methomyl, agricultural chemicals, on pigeon brains. Pigeons were divided into two groups, which were the control and exposure groups. Pigeons of the exposure group were fed with a methomyl-containing diet. After the exposure, the liver, gizzard, heart, lung, muscle, kidney and blood were removed from control and exposure group. These tissues were extracted with methanol, and then analyzed by HPLC. The peaks confirmed the presence of substance derived from methomyl in exposure group’s liver, muscle and blood. Both groups of pigeons were maintained at 80% of normal weight and were experimented with an operant learning. After training of the feeder with hand shaping, the correct reaction ratio of the key pecking was analyzed at a trial time of 20 minutes. In the learning process, the exposure group showed lower correct reaction ratio and post-reinforcement pause compared to the control group. These results may suggest that the exposure of methomyl to pigeons induce disorders in the learning ability. doi:10.1016/j.neures.2009.09.607
P1-n06 Inhibition of fatty achid amide hydrolase accelerates acquisition of cerebellum-dependent eyeblink conditioning Junko Nakayama, Ikuko Oku, Yutaka Kirino, Yasushi Kishimoto Dept. NeuroBiophys, Kagawa Sch. Pharmaceut Sci., Tokushima Bunri. Univ., Sanuki, Japan Recent several studies demonstrated that disruption of CB1 receptor signaling impairs both extinction of hippocampus-dependent memory and acquisition of cerebellum-dependent motor learning. In this study, we tested the hypothesis that elevating brain levels of endogenous cannabinoid anandamide affect cerebellumdependent delay and hippocampus-dependent trace conditionings. C57BL/6J mice treated with FAAH (fatty achid amide hydrolase) inhibitor URB597 (i.p. 1 mg/kg 0.2 ml) did not display any motor discoordination, but did exhibited a significant increase in the rate of delay conditioning. Extinction of CRs were not affected by FAAH inhibitor. On the other hand, FAAH inhibitor had no effect on both acquisi-
Abstracts
N-terminal peptide of mouse Prune2. Human PRUNE2 cDNA encodes a protein that consists of 3,063 amino acids. Western blotting demonstrated the presence endogenous Prune2 protein. Quantitative RT-PCR analysis revealed that the expression of PRUNE2 in brain is the highest among human tissues examined. We demonstrated that Prune2 is a huge protein with the Prune homology domain at the N-terminus and a BCL2/adenovirus E1B interacting protein 2 (BNIP2) and Cdc42GAP homology domain at the C-terminus. doi:10.1016/j.neures.2009.09.601
P1-m20 Jak signaling contributes to Concanavalin A induced C6 glioma cell death Kaoru Nagai, Hideki Yoshida, Yuta Inagawa, Takeo Kubota University of Yamanashi, Japan Because of the difficulty on the treatment of brain tumor including glioma, development of the methods to induce glioma cell death is highly desirable. We examined apoptotic effect of a mannose-binding plant lectin concanavalin A (ConA) on C6 glioma cells. ConA induced C6 cell death dose dependently via carbohydrate binding activity. DNA degradation and caspase-3 activation were observed in ConA treated cells. It indicates that ConA induced apoptotic cell death. Caspase-8 inhibitor significantly reduced the cell death, and a death receptor Fas in C6 cells was recognized by ConA. In addition, JAK inhibitor and gp130-Fc were also inhibited the cell death. Our data suggests that both death receptor and JAK signals contributes ConA induced glioma cell death, and ConA can be utilized to the glioma therapy. doi:10.1016/j.neures.2009.09.602
P1-n01 Interleukin-2 induced Jak-dependent cell death in C6 glioma cells via lectin activity Yuta Inagawa, Takeo Kubota, Kaoru Nagai University of Yamanashi, Japan Gliomas are the most common brain tumors. It was reported that glioma cells secrete molecules disrupting immune systems which contribute tumor elimination. Application of Interleukin-2 (IL-2) was reported to be effective on glioma treatment via stimulating antitumor immune response. However, the direct effect of IL-2 on glioma cells has not been elucidated, yet. In this study, we investigated the effects of IL-2 on C6 glioma cells. IL-2 reduced C6 cell number dose dependently. And the reduction was inhibited by JAK inhibitor, gp130-Fc, and caspase inhibitor. It suggests that IL-2 induced apoptotic cell death in C6 glioma cells via gp130-JAK signal. In addition, since IL-2 was reported to interact with cell surface high-mannose oligosaccharides, we evaluated if IL-2 functions via lectin activity. The haptenic sugar methyl-␣-mannoside significantly blocked the effect of IL-2. Our data suggests that therapeutic glioma treatment with IL-2 directly induced glioma cell death other than stimulating immune response. doi:10.1016/j.neures.2009.09.603
P1-n02 Effects of low-power laser irradiation on proliferation of human-derived glioblastoma A-172 Hideyuki Murayama 1 , Banri Yamanoha 2 , Kei Sadakane 2 , Shinichi Kogure 1 1 Department Bioinformatics, Graduate School of Engineering, Soka University, Tokyo, Japan; 2 Department Environmental Engineering for Symbiosis, Graduate School of Engineering, Soka University, Tokyo, Japan
It has been reported that low-power laser irradiation (LLI) can modulate various biological processes including cell proliferation. The present study was designed to examine effects of 808 nm (30 mW/cm2 ) LLI on the cell proliferation of humanderived glioblastoma A-172 in order to find a possibility for developing an LLI therapy for cancers. According to the conventional cell culture technique, A-172 cells with the same cell density were classified into four groups: control, group 1 (received 20 min dose of LLI), group 2 (40 min), and group 3 (60 min). Cell counting was performed on the PC screen and in some cases with MTT method. In the case of 808 nm LLI (n = 25), it decreased the proliferation ratio as compared with control group: A significant difference (p < 0.01) was obtained between all groups and control. It is concluded that LLI have a potential effect on the cell proliferation of human-derived glioblastoma A-172 with a dose-dependency. doi:10.1016/j.neures.2009.09.604
P1-n03 Electrolytic lesion of the dorsolateral periaqueductal gray attenuates the antinociceptive response of morphine microinjected into the nucleus cuneiformis Leila Ahmad-Molaei, Abbas Haghparast Neurosacience Research Center, Shahid Beheshti University, M.C., It has been indicated that periaqueductal gray (PAG) and nucleus cuneiformis (CnF) play well-established role in a network mediating anti- and pro-nociception. 32 adult male Wistar rats cannulated bilaterally into the CnF, concurrently lesion of dl-PAG was done. Formalin test was carried out to measure pain behavior and antinociceptive effect of morphine microinjected into the CnF (2.5 g/0.3 l saline per side). The results showed that dl-PAG lesion attenuated the effect of morphine microinjected into the CnF in formalin test while dl-PAG lesion solely did not alter basal pain behavior as compared with those in sham-lesion animals. In conclusion, our results suggest the existence of a direct or indirect projection from CnF to the dl-PAG at least at the level of the morphine antinociception in pain modulation. doi:10.1016/j.neures.2009.09.605
P1-n04 Effects of D-serine on fear extinction in mice Shingo Matsuda 1 , Nobuhisa Kanahara 2 , Daisuke Matsuzawa 1 , Ken Nakazawa 1 , Masaomi Iyo 2 , Eiji Shimizu 1 1
Department of Integrative Neurophysiology Graduate School of Medicine, Chiba University, Chiba, Japan; 2 Department of Psychiatry Graduate School of Medicine, Chiba University, Chiba, Japan Molecular mechanisms of fear extinction are not well known. To investigate effects of N-methyl-D-aspartate receptor (NMDA-R) partial agonist, D-serine, on fear extinction we examined cued fear conditioning and extinction paradigm in C57BL/6J mice and measured activation of cytosol and nuclear ERK 1/2 in hippocampus and amygdala. We found that administration of D-serine before extinction facilitated extinction of fear memory at recall tests. In both hippocampus and amygdala, Dserine significantly increased phosphorylation of ERK 1/2. The results suggested that D-serine may facilitate fear extinction through ERK signaling. doi:10.1016/j.neures.2009.09.606
P1-n05 Effects of Methomyl, agricultural chemicals, on learning in pigeons Tomoyuki Kanamatsu, Kentaro Oseki Department Environ. Engi. for Symbiosis, Faculty of Engi., Soka University, Tokyo, Japan The present study was designed to investigate the effects of methomyl, agricultural chemicals, on pigeon brains. Pigeons were divided into two groups, which were the control and exposure groups. Pigeons of the exposure group were fed with a methomyl-containing diet. After the exposure, the liver, gizzard, heart, lung, muscle, kidney and blood were removed from control and exposure group. These tissues were extracted with methanol, and then analyzed by HPLC. The peaks confirmed the presence of substance derived from methomyl in exposure group’s liver, muscle and blood. Both groups of pigeons were maintained at 80% of normal weight and were experimented with an operant learning. After training of the feeder with hand shaping, the correct reaction ratio of the key pecking was analyzed at a trial time of 20 minutes. In the learning process, the exposure group showed lower correct reaction ratio and post-reinforcement pause compared to the control group. These results may suggest that the exposure of methomyl to pigeons induce disorders in the learning ability. doi:10.1016/j.neures.2009.09.607
P1-n06 Inhibition of fatty achid amide hydrolase accelerates acquisition of cerebellum-dependent eyeblink conditioning Junko Nakayama, Ikuko Oku, Yutaka Kirino, Yasushi Kishimoto Dept. NeuroBiophys, Kagawa Sch. Pharmaceut Sci., Tokushima Bunri. Univ., Sanuki, Japan Recent several studies demonstrated that disruption of CB1 receptor signaling impairs both extinction of hippocampus-dependent memory and acquisition of cerebellum-dependent motor learning. In this study, we tested the hypothesis that elevating brain levels of endogenous cannabinoid anandamide affect cerebellumdependent delay and hippocampus-dependent trace conditionings. C57BL/6J mice treated with FAAH (fatty achid amide hydrolase) inhibitor URB597 (i.p. 1 mg/kg 0.2 ml) did not display any motor discoordination, but did exhibited a significant increase in the rate of delay conditioning. Extinction of CRs were not affected by FAAH inhibitor. On the other hand, FAAH inhibitor had no effect on both acquisi-