Metabotropic glutamate receptor 7, coupled with a chemical neurotransmitter L-glutamate, plays an important role in the development of many psychiatric and neurological disorders. To study the biological and genetic m...Metabotropic glutamate receptor 7, coupled with a chemical neurotransmitter L-glutamate, plays an important role in the development of many psychiatric and neurological disorders. To study the biological and genetic mechanism of the mGluR7-related diseases, a physical map covering the full-length mGluR7 genomic sequence has been constructed through seed clone screening and fingerprinting database searching. These BAC clones in the physical map have been sequenced with shotgun strategy and assembled by Phred-Phrap-Consed software; the error rate of the final genoniic sequence is less than 0.01%. mGluR7 spans 880 kb genoniic region, the GC content and repeat content of mGluR7 genoniic sequence are 38% and 37.5% respectively. mGluR7 has a typical 'house-keeping' promoter and consists of 11 exons, with introns ranging from 6 kb to 285 kb. mGluR7a and mGluR7b are two known alternatively splicing variants. Comparing the genomic structures of extracellular domains of mGluR family, their genomic structures can展开更多
Activation of presynaptic group II metabotropic glutamate receptors（mGluR2/3） inhibits drug reward and drug-seeking behavior, but the role of N-acetylaspartylglutamate（NAAG）, an agonist of endogenous mGluR2/3,in h...Activation of presynaptic group II metabotropic glutamate receptors（mGluR2/3） inhibits drug reward and drug-seeking behavior, but the role of N-acetylaspartylglutamate（NAAG）, an agonist of endogenous mGluR2/3,in heroin reward and heroin-seeking behavior remained unclear. Here, we aimed to explore the effects of exogenous NAAG on heroin self-administration and heroinseeking behavior. First, rats were trained to self-administer heroin under a fixed ratio 1（FR1） schedule for 10 days,then received NAAG（50 or 100μg/10 μL in each nostril）in the absence or presence of LY341495（1 mg/kg, i.p.）, an antagonist of mGluR2/3, on day 11 and the effects of NAAG on heroin self-administration under FR1 were recorded for 3 consecutive days. Motivation was assessed in heroin self-administration under a progressive ratio schedule on day 11 in another 5 groups with the same doses of NAAG. Additional rats were withdrawn for 14 days after 14 days of heroin self-administration, then received the same pharmacological pretreatment and were tested for heroin-seeking behaviors induced by heroin priming or cues. The results showed that intranasal administration of NAAG significantly decreased intravenous heroin selfadministration on day 12, but not on day 11. Pretreatment with LY341495 prior to testing on day 12 prevented the inhibitory effect of NAAG on heroin reinforcement. The break-point for reward motivation was significantly reduced by NAAG. Moreover, NAAG also significantly inhibited the heroin-seeking behaviors induced by heroinpriming or cues and these were restored by pretreatment with LY341495. These results demonstrated that NAAG,via activation of presynaptic mGluR2/3, attenuated the heroin reinforcement, heroin motivational value, and heroin-seeking behavior, suggesting that it may be used as an adjunct treatment for heroin addiction.展开更多
Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 group...Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups:control group (C),glutamate group (G),QA group (Q),DCG-IV group (D),L-AP4 group (L) and glutamate+MCPG group (G+M). Cells were cultured under nomoxic condition (95% air,5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes,respectively. G+M group was preincubated with 1mM MCPG for 30 min prior to the stimulation with glutamate. There were 7 time points at 0,4,8,12,16,24 and 48 h in each group except G+M group. Results The expression of VEGF mRNA and protein did not differ significantly among D group,L group and C group. Different from that in C group,the expression of VEGF mRNA and protein could be enhanced both in a dose-dependent and time-dependent manner in G group and Q group. Meanwhile,the enhanced expression of VEGF mRNA and protein in G group was completely suppressed by MCPG after 24 h. Conclusion Glutamate can increase the expression of VEGF mRNA and protein in cultured astrocytes,which may be due to the activation of group I metabotropic glutamate receptors in astrocytes.展开更多
To investigate the expression of interleukin 2 (IL 2), metabotropic glutamate receptor subunit 1 (mGluR1) and estrogen receptor (ER) in neurons of the rat central nervous system (CNS) and identify the coexistence po...To investigate the expression of interleukin 2 (IL 2), metabotropic glutamate receptor subunit 1 (mGluR1) and estrogen receptor (ER) in neurons of the rat central nervous system (CNS) and identify the coexistence possibility of these immune neuro endocrine substances in the central neurons, the tri labeling immunocytochemical technique with different species specific primary antibodies (goat anti IL 2 antibody, rabbit anti mGluR1 antibody and mouse anti ER antibody ) were used to incubate two serial neighbor sections (one for demonstrating IL 2, another for mGluR1 and ER) of the cerebral cortex, medulla oblongata and spinal cord. There were IL 2 , mGluR1 and ER immunoreactivity (IR) positive labeled neurons in the above mentioned central areas. The IL 2 IR production showed brown color, located in the cytoplasm; In the neighbor serial section, the mGluR1 IR, production showed blue black color, located on the cell membrane; the ER IR production also showed brown color, located in the cytoplasm and nuclei. There were mGluR1/ER double labeled cells in the same section, which accounted for about 50 %-60 % of the total single and double labeled neurons. It was identified by projection check of serial neighbor sections that had mGluR1/ER/IL 2 tri labeled cells, which accounted for about 30 % of total mGluR1/ER double labeled neurons. The results indicate that mGluR1, ER and Il 2 can coexist in the same rat central neurons, therefore, providing morphological basis for the theory about immune neuro endocrine network at the cellular level for the first time.展开更多
IL-10 provides trophic and survival effects directly on neurons, promotes axonal outgrowth, and stimulates neuroregeneration. In this study, we analyzed the activities of arginase and nitric oxide synthase (NOS) in sy...IL-10 provides trophic and survival effects directly on neurons, promotes axonal outgrowth, and stimulates neuroregeneration. In this study, we analyzed the activities of arginase and nitric oxide synthase (NOS) in synaptoneurosomes derived from brain cortex of C57BL/6 IL-10 gene-knockout (KO) and wild-type (Wt) mice and determined that the synaptoneurosomes derived from KO mice present lower arginase II activity and lower spermine content than those derived from Wt mice, whereas the basal NOS activity in the KO synaptoneurosomes was higher than that observed in the control synaptoneurosomes. Moreover, our results indicate that the plasma membranes isolated from the KO mice brain exhibit significantly lower spermine-induced enhancement of [3H] MK-801 binding than the plasma membranes from the brain of Wt mice. Glutamate increases the production of nitric oxide (NO) in Wt synaptoneurosomes in a dose-dependent manner, whereas in the KO synaptoneurosomes, this amino acid does not affect the synthesis of NO. The glutamate-dependent acceleration of NO synthesis in Wt synaptoneurosomes was abrogated by LY367385, an antagonist of mGluR1a/b. The western blot analysis of the synaptoneurosomal proteins demonstrates that the expression of the subunits of NMDAR (NMDAR2A and NMDAR2B), the level of NMDAR-bound nNOS and the expression of iNOS are not changed in KO mice and that only the level of mGluR1a/b is markedly reduced in the synaptoneurosomes of KO mice. We conclude that a neuroprotective and neuroregenerative property of IL-10, in addition to its effects on polyamine metabolism and the spermine-dependent modulation of NMDAR, may involve the regulation of mGluR1a/b expression.展开更多
基金This work was supported by the "863" Program of the MOSC (Grant No. 863-J19) the Creative Program of the Chinese Academy of Sciences (Grant No. KSCX1-D4).
文摘Metabotropic glutamate receptor 7, coupled with a chemical neurotransmitter L-glutamate, plays an important role in the development of many psychiatric and neurological disorders. To study the biological and genetic mechanism of the mGluR7-related diseases, a physical map covering the full-length mGluR7 genomic sequence has been constructed through seed clone screening and fingerprinting database searching. These BAC clones in the physical map have been sequenced with shotgun strategy and assembled by Phred-Phrap-Consed software; the error rate of the final genoniic sequence is less than 0.01%. mGluR7 spans 880 kb genoniic region, the GC content and repeat content of mGluR7 genoniic sequence are 38% and 37.5% respectively. mGluR7 has a typical 'house-keeping' promoter and consists of 11 exons, with introns ranging from 6 kb to 285 kb. mGluR7a and mGluR7b are two known alternatively splicing variants. Comparing the genomic structures of extracellular domains of mGluR family, their genomic structures can
基金supported by the National Basic Research Program of China(2015CB553504)the National Natural Science Foundation of China(81471350 and 81671321)the Natural Science Foundation of Ningbo Municipality,Zhejiang Province,China(2015A610193)
文摘Activation of presynaptic group II metabotropic glutamate receptors（mGluR2/3） inhibits drug reward and drug-seeking behavior, but the role of N-acetylaspartylglutamate（NAAG）, an agonist of endogenous mGluR2/3,in heroin reward and heroin-seeking behavior remained unclear. Here, we aimed to explore the effects of exogenous NAAG on heroin self-administration and heroinseeking behavior. First, rats were trained to self-administer heroin under a fixed ratio 1（FR1） schedule for 10 days,then received NAAG（50 or 100μg/10 μL in each nostril）in the absence or presence of LY341495（1 mg/kg, i.p.）, an antagonist of mGluR2/3, on day 11 and the effects of NAAG on heroin self-administration under FR1 were recorded for 3 consecutive days. Motivation was assessed in heroin self-administration under a progressive ratio schedule on day 11 in another 5 groups with the same doses of NAAG. Additional rats were withdrawn for 14 days after 14 days of heroin self-administration, then received the same pharmacological pretreatment and were tested for heroin-seeking behaviors induced by heroin priming or cues. The results showed that intranasal administration of NAAG significantly decreased intravenous heroin selfadministration on day 12, but not on day 11. Pretreatment with LY341495 prior to testing on day 12 prevented the inhibitory effect of NAAG on heroin reinforcement. The break-point for reward motivation was significantly reduced by NAAG. Moreover, NAAG also significantly inhibited the heroin-seeking behaviors induced by heroinpriming or cues and these were restored by pretreatment with LY341495. These results demonstrated that NAAG,via activation of presynaptic mGluR2/3, attenuated the heroin reinforcement, heroin motivational value, and heroin-seeking behavior, suggesting that it may be used as an adjunct treatment for heroin addiction.
基金supported by the National Natural Science Foundation of China(No.30900587)the Natural Science Foundation of Chongqing(No.2009BB5317)the Youth Innovative Foundation of Third Military Medical University,China(No.0656)
基金supported by the National Natural Science Foundation of China (N0.30770673)
文摘Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups:control group (C),glutamate group (G),QA group (Q),DCG-IV group (D),L-AP4 group (L) and glutamate+MCPG group (G+M). Cells were cultured under nomoxic condition (95% air,5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes,respectively. G+M group was preincubated with 1mM MCPG for 30 min prior to the stimulation with glutamate. There were 7 time points at 0,4,8,12,16,24 and 48 h in each group except G+M group. Results The expression of VEGF mRNA and protein did not differ significantly among D group,L group and C group. Different from that in C group,the expression of VEGF mRNA and protein could be enhanced both in a dose-dependent and time-dependent manner in G group and Q group. Meanwhile,the enhanced expression of VEGF mRNA and protein in G group was completely suppressed by MCPG after 24 h. Conclusion Glutamate can increase the expression of VEGF mRNA and protein in cultured astrocytes,which may be due to the activation of group I metabotropic glutamate receptors in astrocytes.
文摘To investigate the expression of interleukin 2 (IL 2), metabotropic glutamate receptor subunit 1 (mGluR1) and estrogen receptor (ER) in neurons of the rat central nervous system (CNS) and identify the coexistence possibility of these immune neuro endocrine substances in the central neurons, the tri labeling immunocytochemical technique with different species specific primary antibodies (goat anti IL 2 antibody, rabbit anti mGluR1 antibody and mouse anti ER antibody ) were used to incubate two serial neighbor sections (one for demonstrating IL 2, another for mGluR1 and ER) of the cerebral cortex, medulla oblongata and spinal cord. There were IL 2 , mGluR1 and ER immunoreactivity (IR) positive labeled neurons in the above mentioned central areas. The IL 2 IR production showed brown color, located in the cytoplasm; In the neighbor serial section, the mGluR1 IR, production showed blue black color, located on the cell membrane; the ER IR production also showed brown color, located in the cytoplasm and nuclei. There were mGluR1/ER double labeled cells in the same section, which accounted for about 50 %-60 % of the total single and double labeled neurons. It was identified by projection check of serial neighbor sections that had mGluR1/ER/IL 2 tri labeled cells, which accounted for about 30 % of total mGluR1/ER double labeled neurons. The results indicate that mGluR1, ER and Il 2 can coexist in the same rat central neurons, therefore, providing morphological basis for the theory about immune neuro endocrine network at the cellular level for the first time.
文摘IL-10 provides trophic and survival effects directly on neurons, promotes axonal outgrowth, and stimulates neuroregeneration. In this study, we analyzed the activities of arginase and nitric oxide synthase (NOS) in synaptoneurosomes derived from brain cortex of C57BL/6 IL-10 gene-knockout (KO) and wild-type (Wt) mice and determined that the synaptoneurosomes derived from KO mice present lower arginase II activity and lower spermine content than those derived from Wt mice, whereas the basal NOS activity in the KO synaptoneurosomes was higher than that observed in the control synaptoneurosomes. Moreover, our results indicate that the plasma membranes isolated from the KO mice brain exhibit significantly lower spermine-induced enhancement of [3H] MK-801 binding than the plasma membranes from the brain of Wt mice. Glutamate increases the production of nitric oxide (NO) in Wt synaptoneurosomes in a dose-dependent manner, whereas in the KO synaptoneurosomes, this amino acid does not affect the synthesis of NO. The glutamate-dependent acceleration of NO synthesis in Wt synaptoneurosomes was abrogated by LY367385, an antagonist of mGluR1a/b. The western blot analysis of the synaptoneurosomal proteins demonstrates that the expression of the subunits of NMDAR (NMDAR2A and NMDAR2B), the level of NMDAR-bound nNOS and the expression of iNOS are not changed in KO mice and that only the level of mGluR1a/b is markedly reduced in the synaptoneurosomes of KO mice. We conclude that a neuroprotective and neuroregenerative property of IL-10, in addition to its effects on polyamine metabolism and the spermine-dependent modulation of NMDAR, may involve the regulation of mGluR1a/b expression.
