高核心岩藻糖基化母乳促进子代肠道双歧杆菌和乳杆菌的优势生长

目的探究母乳蛋白核心岩藻糖基化水平对新生儿肠道菌群结构的影响。方法采用凝集素(aspergillus oryzae lectin,AOL)检测不同母乳样本的蛋白核心岩藻糖基化水平,按照蛋白核心岩藻糖基化水平的高低将母乳样品分为高核心岩藻糖基化组和低核心岩藻糖基化组,采集两组母乳样品对应喂养的婴儿的粪便,利用PCR-变性梯度凝胶电泳(PCR-DGGE)的方法检测两组婴儿粪便中菌群的组成。利用Fut8~(+/-)和Fut8~(+/+)模型小鼠作为母代让其分别哺育野生型子代仔鼠,以进一步阐明核心岩藻糖基化对新生儿肠道菌群的影响。结果母亲/母鼠母乳蛋白岩藻糖基化水平高组与低组相比,婴儿肠道中双歧杆菌菌群丰富度增加,仔鼠肠道内乳杆菌定植增加。结论高核心岩藻糖基化的母乳可以促进子代肠道中双歧杆菌和乳杆菌等益生菌的生长。     

母乳蛋白核心岩藻糖基化有利于新生儿肠道双歧杆菌增长

目的探究母乳蛋白核心岩藻糖基化水平的高低对新生儿肠道双歧杆菌丰富度的影响。方法采用凝集素印记(Lectin Blot, LB)检测不同母乳样品的母乳蛋白核心岩藻糖基化水平,根据蛋白核心岩藻糖基化水平的高低将母乳样品分为两组;采集两组母乳样品对应喂养的新生儿的粪便,通过16S RNA高通量测序的方法检测两组婴儿粪便中菌群的构成。结果母亲母乳蛋白核心岩藻糖基化水平高组相对于低组,其对应的新生儿肠道中双歧杆菌丰富度显著增高。结论高核心岩藻糖基化的母乳可以促进子代肠道中双歧杆菌的优势生长。     

Bmal1时钟基因敲除小鼠的繁育及基因型鉴定

目的对Bmal1基因敲除小鼠进行繁育及基因型进行鉴定,为生物节律研究提供理想的动物模型。方法将引进的Bmal1基因敲除小鼠,以1雄2雌的合笼方式进行饲养繁殖,从仔鼠中提取鼠尾基因组DNA,PCR扩增目的基因片段,琼脂凝胶电泳进行基因结果判定,Western Blot检测心肌组织中Bmal1蛋白表达进行结果验证。结果 Bmal1~(-/-)小鼠与Bmal1~(+/+)小鼠相比,表型除体重外未见明显差异,纯合敲除小鼠丧失繁殖能力; Bmal1~(-/-)小鼠24 h血糖节律改变,Bmal1基因敲除小鼠繁育成功,获得一批基因敲除鼠。结论应用PCR法可成功鉴定Bmal1基因敲除纯合小鼠,PCR扩增法是检测小鼠基因的有效方法。     

口服幽门螺杆菌疫苗候选株对小鼠肠道菌群的影响

目的:考察口服幽门螺杆菌疫苗候选株SH02对小鼠肠道菌群的影响。方法:采用PCR扩增的方法检测疫苗用载体菌侵袭性相关基因缺失状况,用豚鼠角结膜侵袭试验进一步确证其是否具有上皮细胞侵袭能力;用小鼠口服灌胃的方式检测SH02在小鼠体内的组织分布,在肠道的存留时间与活菌数;取小鼠灌胃前(0 d)和灌胃后(2、8 d)的粪便样本,提取基因组DNA,用细菌16S r RNA基因高通量测序的方式,分析评价口服幽门螺杆菌疫苗候选株SH02对小鼠肠道正常菌群的影响。结果:载体菌FWL01侵袭性相关基因缺失,不具有对上皮细胞的侵袭能力;SH02经口服灌胃小鼠后不能侵入机体组织内部,仅在小鼠肠道可以检测到疫苗株活菌,灌胃后24、48 h肠道粪便中活菌数分别为1.19×10~5和2.42×10~3CFU/g,72和96 h在小鼠肠道粪便中没有检测到SH02活菌存在;细菌16S r RNA基因高通量测序与分析结果表明,口服幽门螺杆菌疫苗候选株SH02对小鼠肠道菌群不会产生明显的影响。结论:口服幽门螺杆菌疫苗候选株SH02对小鼠肠道正常菌群没有明显影响。     

东北地区分泌型和非分泌型母亲哺乳期 HMO差异及其对子代肠道菌群的影响

目的分析中国东北地区母亲分泌型和非分泌型基因的分布及其在哺乳期母乳中低聚糖(HMO)种类和含量的差异,探讨这些差异对于子代肠道微生态的影响。方法收集56例母亲在哺乳期第6天母乳样本及其母乳喂养的新生儿在同一天的粪便样本;以质谱分析20种主要HMO的含量;以变性梯度凝胶电泳检测两组新生儿肠道菌群差异。结果所选取的56例母亲其分泌型与非分泌型的比例为43∶13,其中分泌型母亲的岩藻糖基化HMO含量显著高于非分泌型母亲,并且两组新生儿肠道菌群结构也有明显差异。结论不同基因型母亲在哺乳期HMO存在明显差异,而这一差异显著影响了其子代肠道菌群结构。     

Irgm 1参与动脉粥样硬化斑块的形成

目的:探讨免疫相关GTP酶1(Irgm 1)对小鼠血管动脉粥样硬化(AS)斑块形成的影响。方法:高脂饲料喂养野生型(WT)、ApoE~(-/-)Irgm 1~(+/+)和ApoE~(-/-)Irgm1~(+/-)小鼠3个月,建立AS模型;取小鼠主动脉弓,免疫荧光染色方法观察WT和ApoE~(-/-)Irgm 1~(+/+)小鼠血管AS斑块中Irgm 1的表达情况及部位;Western blot方法检测WT和ApoE~(-/-)Irgm 1~(+/+)小鼠血管AS斑块中Irgm 1蛋白表达情况;Q-PCR方法检测WT和ApoE~(-/-)Irgm 1~(+/+)小鼠血管AS斑块中Irgm 1 m RNA表达情况;油红O染色观察ApoE~(-/-)Irgm1~(+/+)和ApoE~(-/-)Irgm1~(+/-)小鼠血管AS斑块形成情况;结果:与WT组相比,ApoE~(-/-)Irgm 1~(+/+)组小鼠主动脉弓AS斑块中Irgm 1+细胞明显增多,Irgm 1+细胞主要位于血管AS斑块的表面;与WT组相比,ApoE~(-/-)Irgm 1~(+/+)组小鼠血管AS斑块中Irgm 1蛋白表达显著增多(P0.001),Irgm 1 m RNA表达显著增多(P0.01);与ApoE~(-/-)Irgm1~(+/-)组相比,ApoE~(-/-)Irgm1~(+/+)组小鼠主动脉弓AS斑块面积显著增大(P0.01);结论:Irgm 1能够促进血管AS斑块的形成。     

肠道菌群及代谢与载脂蛋白E在动脉粥样硬化中的相互作用

目的明确肠道菌群及代谢与载脂蛋白在动脉粥样硬化中的相互作用。方法 C57BL/6J小鼠作为对照组(n=5),载脂蛋白E基因敲除(apolipoprotein E-deficient mice,ApoE~(-/-))小鼠作为动脉粥样硬化模型组(n=8),均为6周龄雄性。两组小鼠均给予高脂饮食,饲养12周后麻醉、摘眼球取血,采用全自动生化分析仪检测血脂,高效液相色谱串联质谱法测定血浆氧化三甲胺(Trimethylamine N-oxid,TMAO)的含量,留取粪便用于肠道菌群16S rRNA V3-V4区域的测序进行菌群鉴定,油红"O"染色主动脉根部确定动脉粥样硬化斑块面积。结果血清学指标:ApoE~(-/-)小鼠较对照组血浆中TMAO(t=-2.87,P0.05)、LDL(t=-11.76,P0.05)、TG(t=-3.56,P0.01)、TC(t=-12.38,P0.01)含量及动脉粥样硬化斑块面积(t=-11.94,P0.01)显著增加,HDL含量降低(t=3.63,P0.01)。肠道菌群测序:两组小鼠随着动脉斑块面积的增加,肠道微生物的丰富度、物种组成及功能预测分析等差异均有统计学意义,还发现ApoE~(-/-)小鼠能够生成TMAO的肠道菌群包括Anaeroplasma、Anaeroplasmatales、Anaeroplasmataceae、Proteus、Paraprevotella和Paraprevotellaceae,而对照组生成TMAO的菌群只有1类,为Anaerotruncus。结论 C57BL/6J小鼠与ApoE~(-/-)小鼠肠道微生物群落的组成、功能及其代谢产物TMAO的血浆含量各不相同,提示ApoE基因缺乏的宿主除导致脂质代谢紊乱外,在动脉粥样硬化形成过程中一定程度上影响着肠道菌群的代谢。     

阿莫西林干预IHFA小鼠及对其成年后肠道菌群的影响

目的利用婴儿菌群人源化小鼠(IHFA小鼠)观察阿莫西林对其干预后及成年后肠道菌群的影响。方法新生Balb/c无菌小鼠接种纯母乳喂养的婴儿粪便获得IHFA小鼠。7~21日龄灌胃给予100mg/kg阿莫西林,对照组给予等量的生理盐水。采用变性梯度凝胶电泳(DGGE)检测小鼠在21日龄及53日龄的肠道菌群。结果 21日龄的阿莫西林处理组IHFA小鼠肠道菌群与正常对照组比较差异有统计学意义(P<0.05);即使在停药后饲养至53日龄的成年小鼠,阿莫西林处理组小鼠肠道菌群仍然存在细微差异。结论哺乳期治疗剂量阿莫西林处理不仅严重干扰小鼠肠道菌群结构,同时导致其成年后肠道菌群不能完全恢复。     

两品系小鼠食物过敏模型的比较

目的比较两种不同品系小鼠食物过敏模型的敏感性和肠道菌群变化的差异,旨在为食物过敏模型的建立提供依据。方法分别对30只4~5周龄BALB/c和KM雌鼠用卵清蛋白(ovalbumin,OVA)致敏建立食物过敏模型,ELISA法检测小鼠血清OVA特异性IgE水平;HE染色观察空肠组织形态;采用DGGE技术检测粪便菌群的变化。结果 (1)30只致敏的BALB/c小鼠中有27只血清OVA特异性IgE水平明显升高(P0.001),而30只致敏的KM小鼠中有21只,且BALB/c小鼠空肠绒毛炎症细胞浸润、上皮脱落及坏死比KM小鼠明显;(2)食物过敏造模后,BALB/c小鼠肠道菌群的改变明显(P0.001),而KM小鼠中仅有均匀度改变显著(P0.05);(3)BALB/c小鼠和KM小鼠对照组肠道菌群的丰富度、Shannon指数及均匀度都有差异。结论 BALB/c小鼠对OVA的敏感性高于KM小鼠,不同品系小鼠肠道菌群结构不同,OVA处理后,BALB/c小鼠菌群的改变比KM小鼠更明显。     

Cidec基因敲除小鼠肠道菌群的特征

目的:探究Cidec敲除(CIDEC-KO)小鼠肠道菌群的结构。方法:随机分别挑选体重相近、2月龄5只野生型和5只Cidec敲除的雄性小鼠,收集两种基因型小鼠经高脂饲料16周喂养前后的新鲜粪便。提取粪便中的细菌基因组,对菌群基因组16S r RNA基因V4高变区进行测序,对数据进行PCoA分析、Alpha多样性分析及LEf Se分析。结果:属水平下的LEf Se分析显示,在普通饲料喂养条件下,Cidec缺失小鼠对比野生型小鼠粪便中PrevotellaceaUCG001属丰度显著上升,Blautia属、Streptococcus属、LachnospiraceaeUCG006属丰度显著下降。与同月龄同高脂喂养的野生型小鼠比,Cidec敲除小鼠粪便中RuminococcaceaeUCG014属丰度显著下降。进一步比较同一种基因型下饮食对肠道菌群的改变,发现在喂养高脂饲料后,某些属的丰度仅在Cidec缺失小鼠中发生显著变化,但是在野生型小鼠中未发现有显著变化,这些属包括:Alistipes属、Bacteroides属、Paraprevotella属、Streptococcus属、LachnospiraceaeUCG006属丰度显著上升。而喂养高脂后,仅在野生型小鼠中发现Peptococcus属和Ruminococcustorquesgroup属丰度的显著上升,以及Tyzzerella3属、Ruminiclostidium6属和A2属丰度的显著下降,在Cidec缺失小鼠粪便中并未发现这些属的丰度有显著变化。结论:高脂诱导下,对比野生型小鼠,某些同代谢综合征正相关的属只在Cidec缺失小鼠肠道菌群中丰度显著上升。     

Core fucosylation regulates epidermal growth factor receptor-mediated intracellular signaling

alpha1,6-Fucosyltransferase (Fut8) catalyzes the transfer of a fucose residue to N-linked oligosaccharides on glycoproteins via an alpha1,6-linkage to form core fucosylation in mammals. We recently found that disruption of the Fut8 gene induces severe growth retardation and early postnatal death. To investigate the molecular mechanism involved, we have established embryonic fibroblasts of Fut8+/+ and Fut8-/-, derived from wild-type and Fut8-null mice, respectively. Interestingly, the epidermal growth factor (EGF)-induced phosphorylation levels of the EGF receptor (EGFR) were substantially blocked in Fut8-/- cells, compared with Fut8+/+ cells, while there are no significant changes in the total activities of tyrosine phosphatase for phosphorylated EGFR between two cells. The inhibition of EGFR phosphorylation was completely restored by re-introduction of the Fut8 gene to Fut8-/- cells. Consistent with this, EGFR-mediated JNK or ERK activation was significantly suppressed in Fut8-/- cells. Finally, we found that the core fucosylation of N-glycans is required for the binding of the EGF to its receptor, whereas no effect was observed for the expression levels of EGFR on the cell surface. Collectively, these results strongly suggest that core fucosylation is essential for EGF receptor-mediated biological functions.     

Gastrointestinal mucins of Fut2-null mice lack terminal fucosylation without affecting colonization by Candida albicans

Posttranslational modification of apomucins by the sequential action of glycosyltransferases is required to produce mature mucins. The Secretor gene (FUT2) encodes an alpha(1,2)fucosyltransferase (EC 2.4.1.69) that catalyzes addition of terminal alpha(1,2)fucose residues on mucins and other molecules in mucosal epithelium. Mutant mice containing targeted replacement of Fut2 with the bacterial reporter gene lacZ were studied to determine the affect of the loss of Fut2 on glycosylation of mucins in the gastrointestinal tract. By whole organ X-gal staining, lacZ activity is prominently expressed in the foveolar pit and chief cells of the glandular stomach, Brunner's glands of the duodenum, and goblet cells in the large intestine of Fut2-LacZ-null mice. Staining with Aleuria aurantia agglutinin demonstrates loss of L-fucosylated epithelial glycans throughout the gastrointestinal tract of Fut2-LacZ-null mice, however, histologic appearance of the tissues appears normal. Analysis of oligosaccharides released from insoluble colonic mucins, largely Muc2, by mass spectrometry shows complete lack of terminal fucosylation of O-linked oligosaccharides in Fut2-LacZ-null mice. Precursor glycans accumulate with no evidence of compensation by other fucosyltransferases or sialyltransferases on mucin glycosylation. Because Candida albicans has been reported to adhere to intestinal mucins creating a potential reservoir associated with vaginitis, Fut2-LacZ-null and wild-type mice were inoculated by gastric lavage with C. albicans. We observe no difference in colonization between genotypes suggesting mucin terminal fucosylation does not significantly influence C. albicans-host interaction in the intestine, highlighting that infections caused by the same organism at different mucosal surfaces are not equal.     

高脂食物对动脉硬化合并类风湿关节炎 小鼠肠道菌群的影响

目的分析高脂食物对动脉硬化合并类风湿关节炎小鼠肠道微生物的影响,了解动脉硬化合并类风湿关节炎小鼠肠道微生物的变化。方法 8周龄ApoE~(-/-)小鼠饲喂高脂食物和普通食物至17周龄来诱发动脉硬化症状,再通过给17周龄ApoE~(-/-)小鼠腹腔注射抗6-磷酸葡萄糖异构酶(glucose-6-phosphate isomerase,GPI)抗体呈阳性的K/BxN血清,从而诱导其产生类风湿关节炎症状。通过Illumina HiSeq平台对各组小鼠粪便进行16S rDNA V4区测序,分析动脉硬化合并类风湿关节炎小鼠肠道微生物的变化。结果 ApoE~(-/-)小鼠饲喂高脂食物后,其血清低密度脂蛋白胆固醇(LDL-C)浓度和血清总胆固醇(TC)浓度均显著升高,主动脉内膜斑块面积比喂普通食物的ApoE~(-/-)小鼠显著增加,表明ApoE~(-/-)小鼠饲喂高脂食物后引起更显著的动脉硬化症状。再通过腹腔注射抗GPI抗体呈阳性的K/BxN血清,各组ApoE~(-/-)小鼠均出现关节肿胀,饲喂高脂食物的ApoE~(-/-)小鼠其踝关节宽度和临床评分(clinical score)低于饲喂普通食物组小鼠。OTU数、Shannon指数和Simpson指数显示高脂食物和K/BxN血清处理组ApoE~(-/-)小鼠肠道菌群多样性降低,Firmicutes/Bacteroidetes值升高,t-test分析显示在属水平上,Prevotellaceae_UCG-001显著降低,Ruminiclostridium_6显著升高。t-test分析和Firmicutes/Bacteroidetes比值显示ApoE~(-/-)小鼠肠道菌群结构紊乱。结论高脂食物使ApoE~(-/-)小鼠的肠道菌群组成和结构发生改变,导致ApoE~(-/-)小鼠的动脉硬化症状加重,类风湿关节炎症状减轻。提示肠道微生物组成和结构的改变,可能与动脉硬化合并类风湿关节炎发病机制相关。     

Haptoglobin and malaria

Haptoglobin gene knockout mice and wild-type controls were infected with Plasmodium berghei ANKA or Plasmodium chabaudi. The peak parasitaemia and parasite burden were higher in Hp-/- mice than in Hp+/+ mice. The increase in spleen weight following malaria infection was smaller in Hp-/- mice than in Hp+/+ animals. The occurrence of cerebral malaria in P. berghei ANKA infection was not different in Hp gene knockout mice and their controls.     

Additional lack of iNOS attenuates diastolic dysfunction in aged ET-1 transgenic mice

Endothelin-1 (ET-1) exhibits potent proinflammatory and profibrotic properties. Moreover, inflammation is a potent stimulus for inducible NO synthase (iNOS), which has been shown to contribute to cardiac injury. We thus hypothesized that ET-1-induced cardiac injury is attenuated by concomitant lack of iNOS. We established crossbred animals of ET-1 transgenic mice (ET+/+) and iNOS knockout mice (iNOS-/-). At 13 months of age, mice were allocated according to their genotype to one of 4 study groups: wild type (WT) controls (n=8); ET-1 transgenic (ET+/+) mice (n=10); iNOS knockout (iNOS-/-) mice (n=7); and crossbred (ET+/+ iNOS-/-) mice (n=15). Left ventricular function was determined in vivo by using a tip catheter. Animals were subsequently euthanized and hearts were harvested for weight assessment and histologic evaluation. No cardiac hypertrophy was present, as evidenced by similar mean cardiac weight and myocyte diameter in all groups. Cardiac perivascular fibrosis was significantly increased in ET+/+ and iNOS-/- groups versus WT, whereas ET+/+ iNOS-/- mice did not differ from WT. Regarding left ventricular function, plasma B-type natriuretic peptide was elevated in ET+/+ and iNOS-/- mice, but again in crossbred animals this effect was blunted. Heart catheterization revealed a significantly increased stiffness constant in both ET-overexpressing groups versus WT, but this increase was significantly attenuated in the ET+/+iNOS-/- group versus the ET+/+ group. Parameters indicating systolic heart failure (EF, cardiac output), however, were not different between all study groups. Our study demonstrates that ET transgenic mice develop left ventricular stiffening with subsequent diastolic dysfunction in a slow, age-dependent manner. Additional knock out of iNOS significantly attenuates cardiac injury. We thus conclude that ET-1-induced cardiac injury is at least partially mediated by iNOS.     

Temporal and spatial dynamics underlying capacitative calcium entry in human colonic smooth muscle

Angiotensin converting enzyme (ACE) has been shown to be involved in regulation of apoptosis in nonintestinal tissues. This study examined the role of ACE in the modulation of intestinal adaptation utilizing ACE knockout mice (ACE-/-). A 60% small bowel resection (SBR) was used, since this model results in a significant increase in intestinal epithelial cell (EC) apoptosis as well as proliferation. Baseline villus height, crypt depth, and intestinal EC proliferation were higher, and EC apoptosis rates were lower in ACE-/- compared with ACE+/+ mice. After SBR, EC apoptosis rates remained significantly lower in ACE-/- compared with ACE+/+ mice. Furthermore, villus height and crypt depth after SBR continued to be higher in ACE-/- mice. The finding of a lower bax-to-bcl-2 protein ratio in ACE-/- mice may account for reduced EC apoptotic rates after SBR in ACE-/- compared with ACE+/+ mice. The baseline higher rate of EC proliferation in ACE-/- compared with ACE+/+ mice may be due to an increase in the expression of several EC growth factor receptors. In conclusion, ACE appears to have an important role in the modulation of intestinal EC apoptosis and proliferation and suggests that the presence of ACE in the intestinal epithelium has a critical role in guiding epithelial cell adaptive response.     

Alpha1,6-fucosyltransferase-deficient mice exhibit multiple behavioral abnormalities associated with a schizophrenia-like phenotype: importance of the balance between the dopamine and serotonin systems

Previously, we reported that α1,6-fucosyltransferase (Fut8)-deficient (Fut8(-/-)) mice exhibit emphysema-like changes in the lung and severe growth retardation due to dysregulation of TGF-β1 and EGF receptors and to abnormal integrin activation, respectively. To study the role of α1,6-fucosylation in brain tissue where Fut8 is highly expressed, we examined Fut8(-/-) mice using a combination of neurological and behavioral tests. Fut8(-/-) mice exhibited multiple behavioral abnormalities consistent with a schizophrenia-like phenotype. Fut8(-/-) mice displayed increased locomotion compared with wild-type (Fut8(+/+)) and heterozygous (Fut8(+/-)) mice. In particular, Fut8(-/-) mice showed strenuous hopping behavior in a novel environment. Working memory performance was impaired in Fut8(-/-) mice as evidenced by the Y-maze tests. Furthermore, Fut8(-/-) mice showed prepulse inhibition (PPI) deficiency. Intriguingly, although there was no significant difference between Fut8(+/+) and Fut8(+/-) mice in the PPI test under normal conditions, Fut8(+/-) mice showed impaired PPI after exposure to a restraint stress. This result suggests that reduced expression of Fut8 is a plausible cause of schizophrenia and related disorders. The levels of serotonin metabolites were significantly decreased in both the striatum and nucleus accumbens of the Fut8(-/-) mice. Likewise, treatment with haloperidol, which is an antipsychotic drug that antagonizes dopaminergic and serotonergic receptors, significantly reduced hopping behaviors. The present study is the first to clearly demonstrate that α1,6-fucosylation plays an important role in the brain, and that it might be related to schizophrenia-like behaviors. Thus, the results of the present study provide new insights into the underlying mechanisms responsible for schizophrenia and related disorders.     

Core fucosylation of μ heavy chains regulates assembly and intracellular signaling of precursor B cell receptors

α1,6-Fucosyltransferase (Fut8) knock-out (Fut8(-/-)) mice showed an abnormality in pre-B cell generation. Membrane assembly of pre-BCR is a crucial checkpoint for pre-B cell differentiation and proliferation in both humans and mice. The assembly of pre-BCR on the cell surface was substantially blocked in the Fut8-knockdown pre-B cell line, 70Z/3-KD cells, and then completely restored by re-introduction of the Fut8 gene to 70Z/3-KD (70Z/3-KD-re) cells. Moreover, loss of α1,6-fucosylation (also called core fucosylation) of μHC was associated with the suppression of the interaction between μHC and λ5. In contrast to Fut8(+/+) CD19(+)CD43(-) cells, the subpopulation expressing the μHC·λ5 complex in the Fut8(-/-) CD19(+)CD43(-) cell fraction was decreased. The pre-BCR-mediated tyrosine phosphorylation of CD79a and activation of Btk were attenuated in Fut8-KD cells, and restored in 70Z/3-KD-re cells. The frequency of CD19(low)CD43(-) cells (pre-B cell enriched fraction) was also reduced in Fut8(-/-) bone marrow cells, and then the levels of IgM, IgG, and IgA of 12-week-old Fut8(-/-) mice sera were significantly lower than those of Fut8(+/+) mice. Our results suggest that the core fucosylation of μHC mediates the assembly of pre-BCR to regulate pre-BCR intracellular signaling and pre-B cell proliferation.     

Alteration of the glutamate and GABA transporters in the hippocampus of the Niemann-Pick disease, type C mouse using proteomic analysis

Niemann-Pick disease type C (NPC) is a fatal autosomal recessive cholesterol disorder characterized by severe progressive neurodegeneration. To unveil the mechanism of neurodegeneration, proteomic and morphological approaches were applied to the hippocampus in NPC -/- mouse. Two-DE was utilized to resolve the hippocampal protein expression profiles of 4- and 8-week-old NPC +/+ and -/- mice. Differentially expressed protein spots were identified by MALDI-TOF MS and database searching. At 4 weeks of age, there was no significant difference in protein profiles between NPC +/+ and -/- mice. However, at the age of 8 weeks, NPC +/+ and -/- mice showed marked difference in protein expressions. Among these, glutamate receptor 2 precursor was identified. The immunohistochemical study on neurotransporters showed that glial GABA transporter (GAT-3) increased in both 4- and 8-week-old NPC -/- mouse and glutamic acid decarboxylase (GAD-6) increased in 8-week-old NPC -/- mouse. Glial glutamate transporter, excitatory amino acids carrier-1 (EAAC1), decreased in 8-week-old NPC -/- mouse. In conclusion, our data may provide insight into the understanding of the basic mechanism through perturbation of protein networks and neurotransporter systems in a single gene knockout model of NPC disease.