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The effect of glutamate on the neuronal differentiation of neural stem cells
Abstracts / Journal of the Neurological Sciences 357 (2015) e235–e254
Presence of an R1 response from the o. oculi muscle during contralateral supraorbital nerve stimulation is unusual. This may indicate cross excitation of the facial nucleus from contralateral trigeminal sensory nerve impulses that are not involved in the usual multi-synaptic pathways responsible for eliciting the R2 contralateral response. Perhaps this is a result of focal CNS demyelination. doi:10.1016/j.jns.2015.08.878
810 WFN15-0143 Miscellaneous Topics 4 The effect of glutamate on the neuronal differentiation of neural stem cells X. Yi, G. Jin, J. Qin, L. Zou. Department of Anatomy and Neurobiology, Nantong University, Nantong, China Background: Our previous study showed that NSCs in peri-injured cortex of adult rats with TBI could differentiate into immature neurons, but they failed to grow up to mature neurons. Objective: To investigate effect of glutamate (Glu) on neuronal differentiation of NSCs. Methods: NSCs were cultured 14 days in four groups: control group, Glu group, Glu + MK-801(0 h) group and Glu + MK-801(24 h) group. Differentiated neurons were observed by DCX or MAP-2 immunofluorescence, cells apoptosis was detected by TUNEL kit. The levels of NR1 mRNA and protein of cells in control and Glu + MK801(24 h) groups were detected by real-time PCR and Western Blot. Results: After 1 day, the number of DCX+ cells in the Glu and Glu + MK-801(24h) group was more than that of control and Glu + MK-801(0h) group. After 7 days many DCX+ cells were still found in the Glu + MK-801(24h) group and more than that of other three groups. After 14 days, many MAP-2+ cells were found in the Glu + MK-801(24h) group and more than that of other three groups. The percentage of TUNEL+ cells in the Glu group peaked at 3rd day then decreased and was still more than that of the other three groups. The expression of NR1 mRNA or protein in the Glu + MK801 group increased gradually and peaked at 7th day then kept in a sable state and was more than that of control group. Conclusion: Glutamate can promote NSCs differentiation into immature neurons then cause cells apoptosis which maybe results from increasing expression of NMDA receptor in the neuronal development. doi:10.1016/j.jns.2015.08.879
811 WFN15-0496 Miscellaneous Topics 4 Association of drd3 gene polymorphisms with parkinson's disease (PD) I. Zhukovaa, S. Ivanovab, V. Alifirovaa, M. Nikitinaa, A. Boikob, O. Fedorenkob, N. Zhukovaa, M. Zhestikovac, N. Bokhanb, M. Titovaa, E. Korolevaa, E. Terskikha, J. Mironovaa, O. Izhboldinaa. aDepartment of neurology and neurosurgery, Siberian state medical university, Tomsk, Russia; bLaboratory of Cellular and Molecular-Biological Investigations, Mental Health Research Institute, Tomsk, Russia; cDepartment of neurology, Novokuznetsk State Postgraduate Institute, Novokuznetsk, Russia Background: PD is one of the most serious and widespread neurodegenerative diseases characterized by chronic progressive course, dysfunction of the basal ganglia and severe disability of
e253
patients. Genetic susceptibility plays a significant role in the development of PD. Objective: To investigate associations between DRD3 dopamine receptor gene tag single nucleotide polymorphisms(SNPs) and the risk of PD development. Material and methods: In this study ten tag SNPs(rs963468, rs2134655, rs9817063, rs324035, rs11721264, rs1800828, rs3773678, rs167770, rs167771, rs7633291) within DRD3 have been genotyped in 143 PD patients and 96 healthy individuals in Russian population. The diagnosis of PD was made using the criteria established by UK Brain Bank. The severity of PD was estimated using Hoehn&Yahr Scale. PD patients and control group were homogeneous by age, race and ratio of men and women(mean age: 69.8 ± 8.6; mean PD duration: 6.9 ± 3.6; women:men = 4:3). Results: Significant associations of 4 polymorphisms(rs11721264, rs3773678, rs167771, rs324035) with PD have been found (Table 1). Conclusions: Our study confirms polymorphic features of dopamine receptor’s genes and their involvement in the pathophysiology in PD. Table 1 The genotype’s distribution of polymorphic variants of DRD3 gene in PD patients and healthy individuals. Polymorphisms
Genotype
PD patients(n,%)
Healthy control subjects(n,%)
χ2
P
rs11721264
AA AG GG AA AG GG AA AG GG AA AC CC
14(11,2%) 54(43,2%) 57(45,6%) 4(2,8%) 30(21%) 109(76,2%) 103(73,6%) 31(22,1%) 6(4,3%) 5(3,5%) 32(22,5%) 105(73,9%)
22(28,9%) 28(36,8%) 26(34,2%) 0(0%) 44(45,4%) 53(54,6%) 60(67,4%) 29(32,6%) 0(0%) 0(0%) 35(36,1%) 62(63,9%)
10,265
0,006
17,846
0,0001
6,368
0,041
8,018
0,018
rs3773678
rs167771
rs324035
doi:10.1016/j.jns.2015.08.880
812 WFN15-0196 Miscellaneous Topics 4 Runx1t1 regulates the neuronal differentiation of neural stem cells from the rat hippocampus L. Zou, G. Jin, J. Qin, X. Yi, J. Shi, H. Li. Department of Human Anatomy and Neuroregeneration, Nantong University, Nantong, China Background: Our previous studies showed expression of Runx1t1 increased during hippocampal neurogenesis. It was hypothesised that Runx1t1 involved the neural development in hippocampus. Purpose: To investigate the function of Runx1t1 in the process of hippocampal neurogenesis. Methods: Expression and localization of Runx1t1 were detected by real-time PCR, Western blot and immunocytochemistry during differentiation of hippocampal neural stem cells(NSCs) in vitro. Runx1t1 in hippocampal NSCs was knocked down by small interfering RNA (siRNA), and Runx1t1 overexpression was induced by LV-Runx1t1 in vitro, to explore the effects of Runx1t1 on the neuronal differentiation of hippocampal NSCs. Results: Expression of Runx1t1 mRNA and proteins in hippocampal NSCs decreased significantly after their differentiation, mainly located in the immature neurons and mature neurons and was not observed in astrocytes. Only 3.6% NSCs differentiated into MAP-2 positive neurons after Runx1t1 knocked down, less than that of the control groups. Moreover, the length and number of neuronal process were also significantly reduced. In contrast, up-regulating
Presence of an R1 response from the o. oculi muscle during contralateral supraorbital nerve stimulation is unusual. This may indicate cross excitation of the facial nucleus from contralateral trigeminal sensory nerve impulses that are not involved in the usual multi-synaptic pathways responsible for eliciting the R2 contralateral response. Perhaps this is a result of focal CNS demyelination. doi:10.1016/j.jns.2015.08.878
810 WFN15-0143 Miscellaneous Topics 4 The effect of glutamate on the neuronal differentiation of neural stem cells X. Yi, G. Jin, J. Qin, L. Zou. Department of Anatomy and Neurobiology, Nantong University, Nantong, China Background: Our previous study showed that NSCs in peri-injured cortex of adult rats with TBI could differentiate into immature neurons, but they failed to grow up to mature neurons. Objective: To investigate effect of glutamate (Glu) on neuronal differentiation of NSCs. Methods: NSCs were cultured 14 days in four groups: control group, Glu group, Glu + MK-801(0 h) group and Glu + MK-801(24 h) group. Differentiated neurons were observed by DCX or MAP-2 immunofluorescence, cells apoptosis was detected by TUNEL kit. The levels of NR1 mRNA and protein of cells in control and Glu + MK801(24 h) groups were detected by real-time PCR and Western Blot. Results: After 1 day, the number of DCX+ cells in the Glu and Glu + MK-801(24h) group was more than that of control and Glu + MK-801(0h) group. After 7 days many DCX+ cells were still found in the Glu + MK-801(24h) group and more than that of other three groups. After 14 days, many MAP-2+ cells were found in the Glu + MK-801(24h) group and more than that of other three groups. The percentage of TUNEL+ cells in the Glu group peaked at 3rd day then decreased and was still more than that of the other three groups. The expression of NR1 mRNA or protein in the Glu + MK801 group increased gradually and peaked at 7th day then kept in a sable state and was more than that of control group. Conclusion: Glutamate can promote NSCs differentiation into immature neurons then cause cells apoptosis which maybe results from increasing expression of NMDA receptor in the neuronal development. doi:10.1016/j.jns.2015.08.879
811 WFN15-0496 Miscellaneous Topics 4 Association of drd3 gene polymorphisms with parkinson's disease (PD) I. Zhukovaa, S. Ivanovab, V. Alifirovaa, M. Nikitinaa, A. Boikob, O. Fedorenkob, N. Zhukovaa, M. Zhestikovac, N. Bokhanb, M. Titovaa, E. Korolevaa, E. Terskikha, J. Mironovaa, O. Izhboldinaa. aDepartment of neurology and neurosurgery, Siberian state medical university, Tomsk, Russia; bLaboratory of Cellular and Molecular-Biological Investigations, Mental Health Research Institute, Tomsk, Russia; cDepartment of neurology, Novokuznetsk State Postgraduate Institute, Novokuznetsk, Russia Background: PD is one of the most serious and widespread neurodegenerative diseases characterized by chronic progressive course, dysfunction of the basal ganglia and severe disability of
e253
patients. Genetic susceptibility plays a significant role in the development of PD. Objective: To investigate associations between DRD3 dopamine receptor gene tag single nucleotide polymorphisms(SNPs) and the risk of PD development. Material and methods: In this study ten tag SNPs(rs963468, rs2134655, rs9817063, rs324035, rs11721264, rs1800828, rs3773678, rs167770, rs167771, rs7633291) within DRD3 have been genotyped in 143 PD patients and 96 healthy individuals in Russian population. The diagnosis of PD was made using the criteria established by UK Brain Bank. The severity of PD was estimated using Hoehn&Yahr Scale. PD patients and control group were homogeneous by age, race and ratio of men and women(mean age: 69.8 ± 8.6; mean PD duration: 6.9 ± 3.6; women:men = 4:3). Results: Significant associations of 4 polymorphisms(rs11721264, rs3773678, rs167771, rs324035) with PD have been found (Table 1). Conclusions: Our study confirms polymorphic features of dopamine receptor’s genes and their involvement in the pathophysiology in PD. Table 1 The genotype’s distribution of polymorphic variants of DRD3 gene in PD patients and healthy individuals. Polymorphisms
Genotype
PD patients(n,%)
Healthy control subjects(n,%)
χ2
P
rs11721264
AA AG GG AA AG GG AA AG GG AA AC CC
14(11,2%) 54(43,2%) 57(45,6%) 4(2,8%) 30(21%) 109(76,2%) 103(73,6%) 31(22,1%) 6(4,3%) 5(3,5%) 32(22,5%) 105(73,9%)
22(28,9%) 28(36,8%) 26(34,2%) 0(0%) 44(45,4%) 53(54,6%) 60(67,4%) 29(32,6%) 0(0%) 0(0%) 35(36,1%) 62(63,9%)
10,265
0,006
17,846
0,0001
6,368
0,041
8,018
0,018
rs3773678
rs167771
rs324035
doi:10.1016/j.jns.2015.08.880
812 WFN15-0196 Miscellaneous Topics 4 Runx1t1 regulates the neuronal differentiation of neural stem cells from the rat hippocampus L. Zou, G. Jin, J. Qin, X. Yi, J. Shi, H. Li. Department of Human Anatomy and Neuroregeneration, Nantong University, Nantong, China Background: Our previous studies showed expression of Runx1t1 increased during hippocampal neurogenesis. It was hypothesised that Runx1t1 involved the neural development in hippocampus. Purpose: To investigate the function of Runx1t1 in the process of hippocampal neurogenesis. Methods: Expression and localization of Runx1t1 were detected by real-time PCR, Western blot and immunocytochemistry during differentiation of hippocampal neural stem cells(NSCs) in vitro. Runx1t1 in hippocampal NSCs was knocked down by small interfering RNA (siRNA), and Runx1t1 overexpression was induced by LV-Runx1t1 in vitro, to explore the effects of Runx1t1 on the neuronal differentiation of hippocampal NSCs. Results: Expression of Runx1t1 mRNA and proteins in hippocampal NSCs decreased significantly after their differentiation, mainly located in the immature neurons and mature neurons and was not observed in astrocytes. Only 3.6% NSCs differentiated into MAP-2 positive neurons after Runx1t1 knocked down, less than that of the control groups. Moreover, the length and number of neuronal process were also significantly reduced. In contrast, up-regulating