NR2B subunits are involved in regulating aging, in particular, age-related learning and memory deficits. We examined 19-month-old NR2B transgenic mice and their littermate controls. First, we detected expression of th...NR2B subunits are involved in regulating aging, in particular, age-related learning and memory deficits. We examined 19-month-old NR2B transgenic mice and their littermate controls. First, we detected expression of the NR2B subunit gene, Grin2b, in the neocortex of transgenic mice using real-time PCR. Next, we used microarrays to examine differences in neocortical gene expression. Pathway and signal-net analyses identified multiple pathways altered in the transgenic mice, in-cluding the P53, Jak-STAT, Wnt, and Notch pathways, as well as regulation of the actin cytoskeleton and neuroactive ligand-receptor interactions. Further signal-net analysis highlighted the P53 and insulin-like growth factor pathways as key regulatory pathways. Our results provide new insight into understanding the molecular mechanisms of NR2B regulated age-related memory storage, normal organismal aging and age-related disease.展开更多
We previously found that the K141N mutation in heat shock protein B8 （HSPB8） was responsible for Charcot-Marie-Tooth disease type 2L in a large Chinese family. The objective of the present study was to generate a tr...We previously found that the K141N mutation in heat shock protein B8 （HSPB8） was responsible for Charcot-Marie-Tooth disease type 2L in a large Chinese family. The objective of the present study was to generate a transgenic mouse model bearing the K141N mutation in the human HSPB8 gene, and to determine whether this K141NHSPB8 transgenic mouse model would manifest the clinical phenotype of Charcot-Marie-Tooth disease type 2L, and consequently be suitable for use in studies of disease pathogenesis. Transgenic mice overexpressing K141N HSPB8 were generated using K141N mutant HSPB8 cDNA cloned into a pCAGGS plasmid driven by a human cytomegalovirus expression system. PCR and western blot analysis confirmed integration of the KI41NHSPB8 gene and widespread expression in tissues of the transgenic mice. The K141N HSPB8 transgenic mice exhibited decreased muscle strength in the hind limbs and impaired motor coordination, but no obvious sensory disturbance at 6 months of age by behavioral assessment. Electrophysiological analysis showed that the compound motor action potential amplitude in the sciatic nerve was significantly decreased, but motor nerve conduction velocity remained normal at 6 months of age. Pathological analysis of the sciatic nerve showed reduced myelinated fiber density, notable axonal edema and vacuolar degeneration in K141N HSPB8 transgenic mice, suggesting axonal involvement in the peripheral nerve damage in these animals. These findings indicate that the KI4mHSPB8 transgenic mouse successfully models Charcot-Marie-Tooth disease type 2L and can be used to study the pathogenesis of the disease.展开更多
目的克隆、表达青海省藏羊源细粒棘球蚴（Echinococcus granulosus）抗原B8/2（Eg Ag B8/2）基因,并进行免疫学鉴定。方法用RT-PCR扩增Eg Ag B8/2基因c DNA,构建原核表达载体pET-Eg Ag B8/2,转化至大肠埃希菌BL21（DE3）中诱导表达,十...目的克隆、表达青海省藏羊源细粒棘球蚴（Echinococcus granulosus）抗原B8/2（Eg Ag B8/2）基因,并进行免疫学鉴定。方法用RT-PCR扩增Eg Ag B8/2基因c DNA,构建原核表达载体pET-Eg Ag B8/2,转化至大肠埃希菌BL21（DE3）中诱导表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹（Western blotting）对诱导表达后的蛋白进行鉴定。结果克隆的Eg Ag B8/2基因长335 bp,序列分析表明与已报道的Eg Ag B8/2 c DNA序列的同源性为98%~100%,原核表达载体pET-Eg Ag B8/2构建正确。诱导表达后,通过SDS-PAGE检测,上清中有大量目的蛋白表达。纯化的蛋白经Western blotting鉴定为目的蛋白。结论成功克隆了青海省藏羊源Eg Ag B8/2基因,构建了原核表达载体,诱导表达后的蛋白为目的蛋白。展开更多
基金supported by the National Natural Science Foundationof China,No.30800312 and 30970368the Specialized Research Fund for the Doctoral Program of Higher Education,No.20070269026+1 种基金the Fundamental Research Fund for the Central Universities,No.78210020Innovation Program of Shanghai Municipal Education Commission,No.10ZZ35
文摘NR2B subunits are involved in regulating aging, in particular, age-related learning and memory deficits. We examined 19-month-old NR2B transgenic mice and their littermate controls. First, we detected expression of the NR2B subunit gene, Grin2b, in the neocortex of transgenic mice using real-time PCR. Next, we used microarrays to examine differences in neocortical gene expression. Pathway and signal-net analyses identified multiple pathways altered in the transgenic mice, in-cluding the P53, Jak-STAT, Wnt, and Notch pathways, as well as regulation of the actin cytoskeleton and neuroactive ligand-receptor interactions. Further signal-net analysis highlighted the P53 and insulin-like growth factor pathways as key regulatory pathways. Our results provide new insight into understanding the molecular mechanisms of NR2B regulated age-related memory storage, normal organismal aging and age-related disease.
基金funded by the National Natural Science Foundation of China,No.81071001,30900805
文摘We previously found that the K141N mutation in heat shock protein B8 （HSPB8） was responsible for Charcot-Marie-Tooth disease type 2L in a large Chinese family. The objective of the present study was to generate a transgenic mouse model bearing the K141N mutation in the human HSPB8 gene, and to determine whether this K141NHSPB8 transgenic mouse model would manifest the clinical phenotype of Charcot-Marie-Tooth disease type 2L, and consequently be suitable for use in studies of disease pathogenesis. Transgenic mice overexpressing K141N HSPB8 were generated using K141N mutant HSPB8 cDNA cloned into a pCAGGS plasmid driven by a human cytomegalovirus expression system. PCR and western blot analysis confirmed integration of the KI41NHSPB8 gene and widespread expression in tissues of the transgenic mice. The K141N HSPB8 transgenic mice exhibited decreased muscle strength in the hind limbs and impaired motor coordination, but no obvious sensory disturbance at 6 months of age by behavioral assessment. Electrophysiological analysis showed that the compound motor action potential amplitude in the sciatic nerve was significantly decreased, but motor nerve conduction velocity remained normal at 6 months of age. Pathological analysis of the sciatic nerve showed reduced myelinated fiber density, notable axonal edema and vacuolar degeneration in K141N HSPB8 transgenic mice, suggesting axonal involvement in the peripheral nerve damage in these animals. These findings indicate that the KI4mHSPB8 transgenic mouse successfully models Charcot-Marie-Tooth disease type 2L and can be used to study the pathogenesis of the disease.
文摘目的克隆、表达青海省藏羊源细粒棘球蚴（Echinococcus granulosus）抗原B8/2（Eg Ag B8/2）基因,并进行免疫学鉴定。方法用RT-PCR扩增Eg Ag B8/2基因c DNA,构建原核表达载体pET-Eg Ag B8/2,转化至大肠埃希菌BL21（DE3）中诱导表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹（Western blotting）对诱导表达后的蛋白进行鉴定。结果克隆的Eg Ag B8/2基因长335 bp,序列分析表明与已报道的Eg Ag B8/2 c DNA序列的同源性为98%~100%,原核表达载体pET-Eg Ag B8/2构建正确。诱导表达后,通过SDS-PAGE检测,上清中有大量目的蛋白表达。纯化的蛋白经Western blotting鉴定为目的蛋白。结论成功克隆了青海省藏羊源Eg Ag B8/2基因,构建了原核表达载体,诱导表达后的蛋白为目的蛋白。
