AIM: To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate(Glu) in Hirschsprung's disease(HSCR).METHODS: Ninety children with HSCR and 50 children without HSCR matched fo...AIM: To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate(Glu) in Hirschsprung's disease(HSCR).METHODS: Ninety children with HSCR and 50 children without HSCR matched for similar nutritional status, age and basal metabolic index were studied. The expression and localization of neuroligin-1 and Glu were assessed using double-labeling immunofluorescence staining of longitudinal muscles with adherent myenteric plexus from the surgically excised colon of children with HSCR. Western blot analysis, quantitative real-time PCR(q RT-PCR) and immunohistochemistry were performed to evaluate the abundance of neuroligin-1 and Glu in different HSCR-affected segments(ganglionic, transitional, and aganglionic segments). Enzyme-linked immunosorbent assay(ELISA) was used to detect and compare serum Glu levels in the long-segment HSCR, short-segment HSCR and non-HSCR samples.RESULTS: Neuroligin-1 and Glu were co-expressed highest to lowest in the ganglionic, transi tional and aganglionic segments based on Western blot(neuroligin-1: 0.177 ± 0.008 vs 0.101 ± 0.006, 0.177 ± 0.008 vs 0.035 ± 0.005, and 0.101 ± 0.006 vs 0.035 ±0.005, P < 0.005; Glu: 0.198 ± 0.006 vs 0.115 ± 0.008, 0.198 ± 0.006 vs 0.040 ± 0.003, and 0.115 ± 0.008 vs 0.040 ± 0.003, P < 0.005) and q RT-PCR(neuroligin-1: 9.58 × 10-5 ± 9.94 × 10-6 vs 2.49 × 10-5 ± 1.38 × 10-6, 9.58 × 10-5 ± 9.94 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, and 2.49 × 10-5 ± 1.38 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, P < 0.005). Serum Glu level was the highest to lowest in the non-HSCR, short-type HSCR and long-type HSCR samples based on ELISA(in nmol/μL, 0.93 ± 0.31 vs 0.57 ± 0.25, 0.93 ± 0.31 vs 0.23 ± 0.16, and 0.57 ± 0.25 vs 0.23 ± 0.16, P < 0.005).CONCLUSION: Neuroligin-1 and Glu may represent new markers of ganglion cells, whose expression may correlate with the pathogenesis, diagnosis, differential diagnosis or classification of HSCR.展开更多
目的研究先天性巨结肠症(HD)患儿各段肠管中Neuroligin2基因的表达情况,探讨Neuroligin2表达与HD发生的可能关系。方法选取山东大学齐鲁医院小儿外科行手术治疗的HD患儿结肠标本共42例,将HD患儿正常段设为正常对照组,移行段及狭窄段设...目的研究先天性巨结肠症(HD)患儿各段肠管中Neuroligin2基因的表达情况,探讨Neuroligin2表达与HD发生的可能关系。方法选取山东大学齐鲁医院小儿外科行手术治疗的HD患儿结肠标本共42例,将HD患儿正常段设为正常对照组,移行段及狭窄段设为实验组,应用Western blotting、实时荧光定量聚合酶链式反应(qRT-PCR)及免疫组织化学染色方法观察Neuroligin2在各肠段的表达情况。结果 Western blotting结果显示Neuroligin2蛋白在狭窄段相对表达量为0.012±0.001,明显少于移行段和正常段(0.039±0.001,0.057±0.002;P<0.05)。qRT-PCR结果显示,Neuroligin2的mRNA水平与蛋白表达趋势一致。免疫组织化学染色示HD患儿正常段肠管的肌间肠神经节细胞中有Neuroligin2强阳性细胞的表达,而在狭窄段肠管中弱阳性表达,移行段肠管可见Neuroligin2阳性细胞的表达。结论 HD患儿狭窄段肠管中Neuroligin2的表达缺失可能与HD的发生有关。展开更多
Neuroligins(NLs) are postsynaptic cell-adhesion proteins that play important roles in synapse formation and the excitatory-inhibitory balance. They have been associated with autism in both human genetic and animal mod...Neuroligins(NLs) are postsynaptic cell-adhesion proteins that play important roles in synapse formation and the excitatory-inhibitory balance. They have been associated with autism in both human genetic and animal model studies, and affect synaptic connections and synaptic plasticity in several brain regions. Yet current research mainly focuses on pyramidal neurons, while the function of NLs in interneurons remains to be understood. To explore the functional difference among NLs in the subtypespecific synapse formation of both pyramidal neurons and interneurons, we performed viral-mediated shRNA knockdown of NLs in cultured rat cortical neurons and examined the synapses in the two major types of neurons. Our results showed that in both types of neurons, NL1 and NL3 were involved in excitatory synapse formation, and NL2 in GABAergic synapse formation. Interestingly, NL1 affectedGABAergic synapse formation more specifically than NL3,and NL2 affected excitatory synapse density preferentially in pyramidal neurons. In summary, our results demonstrated that different NLs play distinct roles in regulating the development and balance of excitatory and inhibitory synapses in pyramidal neurons and interneurons.展开更多
Carboxylesterases are enzymes widely distributed within living organisms. In insects, they have been mainly involved in dietary metabolism and detoxification function. Interestingly, several members of this family cal...Carboxylesterases are enzymes widely distributed within living organisms. In insects, they have been mainly involved in dietary metabolism and detoxification function. Interestingly, several members of this family called carboxylesterase-like ad- hesion molecules （CLAMs） have lost their catalytic properties and are mainly involved in neuro/developmental functions. CLAMs include gliotactins, neurotactins, glutactins, and neuroligins. The latter have for binding partner the neurexin. In insects, the function of these proteins has been mainly studied in Drosophila central nervous system or neuro- muscular junction. Some studies suggested a role of neuroligins and neurexin in sensory processing but CLAM expression within sensory systems has not been investigated. Here, we reported the identification of 5 putative CLAMs expressed in the olfactory system of the model pest insect Spodoptera littoralis. One neuroligin, Slnlg4-yll and its putative binding partner neurexin Slnrxl were the most expressed in the antennae and were surprisingly associated with olfactory sensilla. In addition, both transcripts were upregulated in male antennae after mating, known to modulate the sensitivity of the peripheral olfactory system in S. littoralis, suggesting that these molecules could be involved in sensory plasticity.展开更多
Neuroligins （Nlgs） are transmembrane cell adhesion molecules playing essential roles in synapse development and function. Genetic mutations in neuroUgin genes have been linked with some neurodevelopmental disorders ...Neuroligins （Nlgs） are transmembrane cell adhesion molecules playing essential roles in synapse development and function. Genetic mutations in neuroUgin genes have been linked with some neurodevelopmental disorders such as autism. These mutated Nlgs are mostly retained in the endoplasmic reticulum （ER）. However, the mechanisms underlying normal Nlg maturation and trafficking have remained largely unknown. Here, we found that Drosophila neuroligin 2 （DNlg2） undergoes proteolytic cleavage in the ER in a variety of Drosophila tissues throughout developmental stages. A region encompassing Y642-T698 is required for this process. The immature non-cleavable DNtg2 is retained in the ER and non-functionaL The C-terminal fragment of DNlg2 instead of the full-length or non-cleavable DNIg2 is able to rescue neuromuscular junction defects and GluRIIB reduction induced by dnlg2 deletion. Intriguingly, the autism-associated R598C mutation in DNIg2 leads to similar marked defects in DNIg2 proteo- lytic process and ER export, revealing a potential role of the improper Nlg cleavage in autism pathogenesis. Collectively, our find- ings uncover a specific mechanism that controls DNIg2 maturation and trafficking via proteolytic cleavage in the ER, suggesting that the perturbed proteolytic cleavage of Nlgs likely contributes to autism disorder.展开更多
Gamma-aminobutyric acid(GABA)is the major inhibitory neurotransmitter in the brain.As one of several types of endogenous receptors,GABA_(A)receptors have been shown to be essential in most,if not all,aspects of brain ...Gamma-aminobutyric acid(GABA)is the major inhibitory neurotransmitter in the brain.As one of several types of endogenous receptors,GABA_(A)receptors have been shown to be essential in most,if not all,aspects of brain functioning,including neural development and information processing.Mutations in genes encoding GABA_(A)receptors and alterations in the function of GABA_(A)receptors are associated with many neurologic diseases,and GABA_(A)receptors have been clinically targeted by many drugs,such as benzodiazepines and general anesthetics.Extensive studies have revealed a number of intracellular chaperons/interactions for GABA_(A)receptors,providing a protein-protein network in regulating the trafficking and location of GABA_(A)receptors in the brain.Recently,neurexins and neuroligins,two families of transmembrane proteins present at neurological synapses,are implicated as new partners to GABA_(A)receptors.These works shed new light on the synaptic regulation of GABA_(A)receptor activity.Here,we summarized the proteins that were implicated in the function of GABA_(A)receptors,including neurexins and neuroligins.展开更多
目的:探讨肿瘤微环境中神经元对胶质瘤细胞迁移能力的影响。方法:分离培养胚胎18 d(E18)大鼠神经元,利用表达靶向神经连接蛋白3(NLGN3)分子shRNA的重组慢病毒(Lv-NLGN3-shRNA)感染神经元,real time RT-PCR和酶联免疫吸附试验(ELISA)检测...目的:探讨肿瘤微环境中神经元对胶质瘤细胞迁移能力的影响。方法:分离培养胚胎18 d(E18)大鼠神经元,利用表达靶向神经连接蛋白3(NLGN3)分子shRNA的重组慢病毒(Lv-NLGN3-shRNA)感染神经元,real time RT-PCR和酶联免疫吸附试验(ELISA)检测NLGN3的表达,收集神经元培养上清与胶质瘤细胞系U251细胞共培养,Transwell实验检测U251细胞迁移能力,Western Blot方法检测U251细胞中哺乳动物雷帕霉素靶蛋白(mTOR)和基质金属蛋白酶-9(MMP-9)的表达。结果:Lv-NLGN3-shRNA重组慢病毒感染能够降低大鼠原代神经元中NLGN3 mRNA和蛋白的的表达,神经元培养上清与U251细胞共培养可以增加后者的迁移能力并上调mTOR和MMP-9的表达。然而,NLGN3表达被Lv-NLGN3-shRNA抑制后,培养上清对U251细胞促迁移能力下降,同时mTOR和MMP-9表达降低。结论:大鼠原代神经元可以通过NLGN3/mTOR/MMP-9信号通路增强U251细胞的迁移能力,这一途径可能是胶质瘤细胞转移的机制之一。展开更多
目的:探讨N-甲基-D-天冬氨酸(NMDA)对大鼠神经元来源的细胞系H19-7细胞中神经连接蛋白3(NLGN3)表达的影响及分子机制。方法:分别利用NMDA、NMDA受体(NMDAR)特异性拮抗剂D-AP5和β-catenin特异性siRNA处理大鼠海马神经元细胞系H19-7细胞...目的:探讨N-甲基-D-天冬氨酸(NMDA)对大鼠神经元来源的细胞系H19-7细胞中神经连接蛋白3(NLGN3)表达的影响及分子机制。方法:分别利用NMDA、NMDA受体(NMDAR)特异性拮抗剂D-AP5和β-catenin特异性siRNA处理大鼠海马神经元细胞系H19-7细胞,通过Western Blot和real time RT-PCR检测β-连环蛋白(β-catenin)、NLGN3及c-Myc的表达变化。结果:NMDA刺激后H19-7细胞中β-catenin蛋白上调,与此同时NLGN3和c-Myc表达增加;使用D-AP5阻断NMDAR以及特异性siRNA沉默β-catenin均能抑制NLGN3的表达。结论:NMDA能够通过Wnt/β-catenin通路促进大鼠神经元中NLGN3表达。展开更多
基金Supported by National Natural Science Foundation of China,No.81270720 and No.81471487
文摘AIM: To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate(Glu) in Hirschsprung's disease(HSCR).METHODS: Ninety children with HSCR and 50 children without HSCR matched for similar nutritional status, age and basal metabolic index were studied. The expression and localization of neuroligin-1 and Glu were assessed using double-labeling immunofluorescence staining of longitudinal muscles with adherent myenteric plexus from the surgically excised colon of children with HSCR. Western blot analysis, quantitative real-time PCR(q RT-PCR) and immunohistochemistry were performed to evaluate the abundance of neuroligin-1 and Glu in different HSCR-affected segments(ganglionic, transitional, and aganglionic segments). Enzyme-linked immunosorbent assay(ELISA) was used to detect and compare serum Glu levels in the long-segment HSCR, short-segment HSCR and non-HSCR samples.RESULTS: Neuroligin-1 and Glu were co-expressed highest to lowest in the ganglionic, transi tional and aganglionic segments based on Western blot(neuroligin-1: 0.177 ± 0.008 vs 0.101 ± 0.006, 0.177 ± 0.008 vs 0.035 ± 0.005, and 0.101 ± 0.006 vs 0.035 ±0.005, P < 0.005; Glu: 0.198 ± 0.006 vs 0.115 ± 0.008, 0.198 ± 0.006 vs 0.040 ± 0.003, and 0.115 ± 0.008 vs 0.040 ± 0.003, P < 0.005) and q RT-PCR(neuroligin-1: 9.58 × 10-5 ± 9.94 × 10-6 vs 2.49 × 10-5 ± 1.38 × 10-6, 9.58 × 10-5 ± 9.94 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, and 2.49 × 10-5 ± 1.38 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, P < 0.005). Serum Glu level was the highest to lowest in the non-HSCR, short-type HSCR and long-type HSCR samples based on ELISA(in nmol/μL, 0.93 ± 0.31 vs 0.57 ± 0.25, 0.93 ± 0.31 vs 0.23 ± 0.16, and 0.57 ± 0.25 vs 0.23 ± 0.16, P < 0.005).CONCLUSION: Neuroligin-1 and Glu may represent new markers of ganglion cells, whose expression may correlate with the pathogenesis, diagnosis, differential diagnosis or classification of HSCR.
基金supported by National Basic Research Development Program of China(No.2010CB912002)the Natural ScienceFoundation of Zhejiang Province,China(No.LY12C09001)Fundamental Research Funds for the Central Universities of China(No.2012QNA7007)
文摘目的研究先天性巨结肠症(HD)患儿各段肠管中Neuroligin2基因的表达情况,探讨Neuroligin2表达与HD发生的可能关系。方法选取山东大学齐鲁医院小儿外科行手术治疗的HD患儿结肠标本共42例,将HD患儿正常段设为正常对照组,移行段及狭窄段设为实验组,应用Western blotting、实时荧光定量聚合酶链式反应(qRT-PCR)及免疫组织化学染色方法观察Neuroligin2在各肠段的表达情况。结果 Western blotting结果显示Neuroligin2蛋白在狭窄段相对表达量为0.012±0.001,明显少于移行段和正常段(0.039±0.001,0.057±0.002;P<0.05)。qRT-PCR结果显示,Neuroligin2的mRNA水平与蛋白表达趋势一致。免疫组织化学染色示HD患儿正常段肠管的肌间肠神经节细胞中有Neuroligin2强阳性细胞的表达,而在狭窄段肠管中弱阳性表达,移行段肠管可见Neuroligin2阳性细胞的表达。结论 HD患儿狭窄段肠管中Neuroligin2的表达缺失可能与HD的发生有关。
基金supported by grants from the National Natural Science Foundation of China(31571049 and81561168022)the National Basic Research Program of China(2015CB910801)+2 种基金Zhejiang Provincial Natural Science Foundation of China(LR19H090001 and LD19H090002)a joint grant from the National Natural Science Foundation of China and the Research Grants Council of Hong Kong,China(8151101104 and N_HKUST625/15)Fundamental Research Funds for the CentralUniversities of China
文摘Neuroligins(NLs) are postsynaptic cell-adhesion proteins that play important roles in synapse formation and the excitatory-inhibitory balance. They have been associated with autism in both human genetic and animal model studies, and affect synaptic connections and synaptic plasticity in several brain regions. Yet current research mainly focuses on pyramidal neurons, while the function of NLs in interneurons remains to be understood. To explore the functional difference among NLs in the subtypespecific synapse formation of both pyramidal neurons and interneurons, we performed viral-mediated shRNA knockdown of NLs in cultured rat cortical neurons and examined the synapses in the two major types of neurons. Our results showed that in both types of neurons, NL1 and NL3 were involved in excitatory synapse formation, and NL2 in GABAergic synapse formation. Interestingly, NL1 affectedGABAergic synapse formation more specifically than NL3,and NL2 affected excitatory synapse density preferentially in pyramidal neurons. In summary, our results demonstrated that different NLs play distinct roles in regulating the development and balance of excitatory and inhibitory synapses in pyramidal neurons and interneurons.
文摘Carboxylesterases are enzymes widely distributed within living organisms. In insects, they have been mainly involved in dietary metabolism and detoxification function. Interestingly, several members of this family called carboxylesterase-like ad- hesion molecules （CLAMs） have lost their catalytic properties and are mainly involved in neuro/developmental functions. CLAMs include gliotactins, neurotactins, glutactins, and neuroligins. The latter have for binding partner the neurexin. In insects, the function of these proteins has been mainly studied in Drosophila central nervous system or neuro- muscular junction. Some studies suggested a role of neuroligins and neurexin in sensory processing but CLAM expression within sensory systems has not been investigated. Here, we reported the identification of 5 putative CLAMs expressed in the olfactory system of the model pest insect Spodoptera littoralis. One neuroligin, Slnlg4-yll and its putative binding partner neurexin Slnrxl were the most expressed in the antennae and were surprisingly associated with olfactory sensilla. In addition, both transcripts were upregulated in male antennae after mating, known to modulate the sensitivity of the peripheral olfactory system in S. littoralis, suggesting that these molecules could be involved in sensory plasticity.
文摘Neuroligins （Nlgs） are transmembrane cell adhesion molecules playing essential roles in synapse development and function. Genetic mutations in neuroUgin genes have been linked with some neurodevelopmental disorders such as autism. These mutated Nlgs are mostly retained in the endoplasmic reticulum （ER）. However, the mechanisms underlying normal Nlg maturation and trafficking have remained largely unknown. Here, we found that Drosophila neuroligin 2 （DNlg2） undergoes proteolytic cleavage in the ER in a variety of Drosophila tissues throughout developmental stages. A region encompassing Y642-T698 is required for this process. The immature non-cleavable DNtg2 is retained in the ER and non-functionaL The C-terminal fragment of DNlg2 instead of the full-length or non-cleavable DNIg2 is able to rescue neuromuscular junction defects and GluRIIB reduction induced by dnlg2 deletion. Intriguingly, the autism-associated R598C mutation in DNIg2 leads to similar marked defects in DNIg2 proteo- lytic process and ER export, revealing a potential role of the improper Nlg cleavage in autism pathogenesis. Collectively, our find- ings uncover a specific mechanism that controls DNIg2 maturation and trafficking via proteolytic cleavage in the ER, suggesting that the perturbed proteolytic cleavage of Nlgs likely contributes to autism disorder.
基金supported by the National Basic Research Program of China(973 program No.2011CB809102 to C.Z.)“985”Research Foundation of Peking University(to C.Z.).
文摘Gamma-aminobutyric acid(GABA)is the major inhibitory neurotransmitter in the brain.As one of several types of endogenous receptors,GABA_(A)receptors have been shown to be essential in most,if not all,aspects of brain functioning,including neural development and information processing.Mutations in genes encoding GABA_(A)receptors and alterations in the function of GABA_(A)receptors are associated with many neurologic diseases,and GABA_(A)receptors have been clinically targeted by many drugs,such as benzodiazepines and general anesthetics.Extensive studies have revealed a number of intracellular chaperons/interactions for GABA_(A)receptors,providing a protein-protein network in regulating the trafficking and location of GABA_(A)receptors in the brain.Recently,neurexins and neuroligins,two families of transmembrane proteins present at neurological synapses,are implicated as new partners to GABA_(A)receptors.These works shed new light on the synaptic regulation of GABA_(A)receptor activity.Here,we summarized the proteins that were implicated in the function of GABA_(A)receptors,including neurexins and neuroligins.
文摘目的:探讨N-甲基-D-天冬氨酸(NMDA)对大鼠神经元来源的细胞系H19-7细胞中神经连接蛋白3(NLGN3)表达的影响及分子机制。方法:分别利用NMDA、NMDA受体(NMDAR)特异性拮抗剂D-AP5和β-catenin特异性siRNA处理大鼠海马神经元细胞系H19-7细胞,通过Western Blot和real time RT-PCR检测β-连环蛋白(β-catenin)、NLGN3及c-Myc的表达变化。结果:NMDA刺激后H19-7细胞中β-catenin蛋白上调,与此同时NLGN3和c-Myc表达增加;使用D-AP5阻断NMDAR以及特异性siRNA沉默β-catenin均能抑制NLGN3的表达。结论:NMDA能够通过Wnt/β-catenin通路促进大鼠神经元中NLGN3表达。
