C57BL/6小鼠不同组织器官中NKT细胞的比较

目的比较C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结中NKT细胞的含量、亚型和功能的特点。方法分离正常C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结的淋巴细胞,利用细胞表面分子染色的方法,观察不同组织器官中CD3+NK1.1+NKT细胞及其亚型的含量;淋巴细胞经过PMA和离子霉素刺激后,应用细胞内细胞因子染色的方法,通过流式细胞仪观察NKT细胞IFN-γ、IL-4、IL-9和IL-17的产生情况。结果肝脏中NKT细胞的含量为(25.2±12)%,显著高于肺脏、脾脏和肠系膜淋巴结。肝脏、脾脏和肠系膜淋巴结中NKT细胞以CD4+细胞亚群为主,而肺脏中NKT细胞以CD4-CD8-亚群为主,同时肠系膜淋巴结的NKT细胞中存在CD4+CD8+亚群。不同组织器官中NKT细胞IFN-γ、IL-4、IL-9和IL-17产生的能力有差别。结论 C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结的NKT细胞在含量、表型和功能方面可能存在明显的差异。     

Id2基因过表达对C57BL/6小鼠免疫表型的影响

目的：建立分化抑制因子2(ID2)过表达和CD45.1,CD45.2嵌合体小鼠模型,探讨ID2过表达(ID2^o/o)对C57BL/6小鼠免疫表型的影响。方法：利用基因编辑技术获得ID2^o/o C57BL/6模型小鼠,采用PCR、琼脂糖凝胶电泳和Western blot对小鼠进行基因型鉴定。Western blot检测ID2^o/o和WT小鼠脾脏细胞中各种ID蛋白(ID1～ID4)表达情况。流式细胞术绝对定量方法检测WT和ID2^o/o小鼠脾脏和外周血细胞中各免疫细胞数量。构建骨髓1∶1竞争性嵌合体小鼠模型,以同样方法检测CD45.1⁺和CD45.2⁺白细胞及相应免疫细胞数量。结果：PCR和Western blot结果表明ID2^o/o小鼠构建成功。在ID2^o/o小鼠脾细胞中,4种ID蛋白的表达量均显著高于WT小鼠。流式细胞分析结果显示,Id2基因过表达使C57BL/6小鼠脾脏和外周血中的中性粒细胞、巨噬细胞、树突状细胞...     

约氏疟原虫感染小鼠脾脏B细胞和NK细胞及其细胞因子的检测

为研究约氏疟原虫感染小鼠脾脏不同免疫细胞及其细胞因子的水平变化,将C57BL/6小鼠分为感染组和正常组,分别经小鼠尾静脉注射约氏疟原虫和生理盐水,8d后处死感染组和正常组小鼠并分离脾脏,制备单细胞悬液,然后利用流式细胞术检测小鼠脾脏B细胞和NK细胞及其表达的细胞因子IFN-y、IL-12、IL-10的水平变化情况.结果...     

C57BL/6小鼠γδT细胞的组织器官分布和功能特点

目的 比较C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结中γδT细胞占所分离的淋巴细胞及其CD3+T细胞的百分比、表型和功能的特点.方法 分离正常C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结的淋巴细胞,应用细胞表面分子染色的方法,使用流式细胞仪观察γδT细胞占从不同组织分离的淋巴细胞及CD3+T细胞的百分比及其表型的特点.细胞经PMA和离子霉素刺激后,应用细胞内细胞因子染色的方法,通过流式细胞仪观察γδT细胞产生IFN-γ、IL-4、IL-9和IL-17细胞因子的情况.结果 γδT细胞占分离淋巴细胞中的百分含量在肝脏明显高于肺脏、脾脏和肠系膜淋巴结(P＜0.05),而γδT细胞在肠系膜淋巴结CD3+T细胞的百分含量要显著的低于其他脏器(P＜0.05).不同组织器官中γδT细胞以CD4-CD8-表型为主,还存在少量CD8+ γδT细胞.在肠系膜淋巴结γδT细胞中CD4+细胞的量明显高于其他器官,且明显可见一群CD4+ CD8+细胞.不同组织中γδT细胞中IL-17+的细胞量要明显高于IFN-γ+和IL-4+细胞.γδT细胞基本不分泌IL-9.肺脏的γδT细胞分泌细胞因子的能力最强,其IL-17+细胞的量达到(26.6±12.1)％.IFN-γ+细胞的量在肝脏和肺脏中较高,分别为(1.36±0.37)％和(1.6±0.7)％.结论 C57BL/6小鼠肝脏、肺脏、脾脏和肠系膜淋巴结γδT细胞的含量、表型和功能方面存在显著性差异.     

失血性休克对脾脏细胞因子和花生四烯酸代谢的影响

目的：探讨脾脏在失血性休克时对脾脏细胞因子和花生四烯酸代谢的调节作用,方法：分别失血性休克家犬和家兔动脉血、脾静脉血和脾组织均浆放射免疫法测定其IL－1、IL－2、IL－6、IL－8、TNF、TXB2和6KPGF1α含量.结果：1．失血性休克后动脉血浆中IL－1、IL－6、IL－8、TNF、含量显著降低,而IL－2、TXB2和6KPGF1α含量明显增高,2．失血性休克后脾静脉血浆中IL－1、IL－2,IL－6,IL－8、TNF、6KPGF1α含量明显降低,仅TXB2含量明显增高,而休克治疗后IL－8TNF、6KPGF1α含量均明显产高;3．失血性休克后脾组织中IL－1和TXB2含量降低,IL－2、IL－6、TNF、6KPGF1α含量明显增高;4．切脾对TXB2生成无影响,而对6KPGF1α的生成有一定影响,结论：脾及对失血性休克时的多种功能因子的生成和释放有一定作用,对花生四烯酸代谢也有一定影响。     

短暂性脑缺血对C57BL/6小鼠海马区神经发生的影响

目的:利用小鼠急性全脑缺血模型观察急性脑缺血对海马神经发生的影响。方法:将C57BL/6雄性小鼠(6~8周)随机分为脑缺血组和假手术组。采用双侧颈动脉结扎法建立短暂性全脑缺血模型;利用HE染色观察脑缺血后不同时间(1周,2周)海马区形态学变化;采用免疫组织荧光染色观察缺血后海马区caspase-3表达变化;利用Ed U染色观察缺血3 d、7 d、14 d和28 d后,海马区神经干细胞增殖变化;利用免疫组织荧光染色观察缺血3 d、7 d、14 d和28 d后,海马区nestin、胶质纤维酸性蛋白(GFAP)及少突胶质细胞特异蛋白(OSP)等蛋白表达变化,判断脑缺血对海马区神经干细胞分化的影响。结果:脑缺血7 d后,小鼠海马CA1区神经元数目减少,caspase-3阳性细胞增加(P0.05)。海马DG区Ed U阳性细胞数量在第3 d开始增加(P0.05),第1周达到峰值(P0.05);海马CA1区Ed U阳性细胞数量在缺血后1周开始增加(P0.05),缺血2周后逐渐降低。海马DG区nestin、GFAP及OSP阳性细胞数量均在缺血3 d后开始增加(P0.05),缺血1周后达到峰值(P0.05),2周后逐渐降低。海马CA1区GFAP及OSP阳性细胞数量在缺血3 d后开始增加(P0.05),缺血1周后达到峰值(P0.05),2周后逐渐降低。结论:脑缺血激活了海马DG区神经干细胞,使干细胞增殖,并向神经元、星形胶质细胞、少突胶质细胞分化,同时逐渐向CA1区迁移。     

TLR7介导日本血吸虫感染C57BL/6小鼠脾脏的T细胞反应

目的 探究日本血吸虫感染后小鼠脾脏TLR7对T细胞反应的调节作用。方法 使用日本血吸虫感染6周的C57BL/6小鼠（WT）和TLR7-KO小鼠,并设立相应对照组。然后观察组织病变情况,分离脾脏淋巴细胞,运用qRT-PCR、流式细胞术检测脾脏中CD4⁺T和CD8⁺T细胞数量改变以及TLR7分子表达;进一步通过流式细胞术检测WT和TLR7-KO小鼠感染前后CD4⁺T和CD8⁺T细胞CD25、CD69、CXCR5、CD40L等活化表型的变化;经PMA和离子霉素的刺激,使用细胞内细胞因子染色的方法检测CD4⁺T和CD8⁺T细胞IFN-γ、IL-4分泌情况。结果 日本血吸虫感染后,小鼠肝脏和脾脏明显增大,脾脏中CD4⁺T和CD8⁺T细胞发生大量聚集;感染后CD4⁺T和CD8⁺T细胞多种活化及功能相关指标均显著上升,且IL-4⁺CD4⁺T...     

非酒精性脂肪肝C57BL/6小鼠模型的建立

 BACKGROUND: The establishment of a safe, reliable and easily repeatable mouse model of nonalcoholic fatty liver disease is the prerequisite for the study of the diagnosis and treatment of the disease.     

C57BL/6小鼠皮肤毛囊发育的实验研究

目的探讨胎鼠,乳鼠和成鼠毛囊发育的规律。方法石蜡和冰冻超薄切片,采用HE染色,AP活性染色,油红O染色,凋亡细胞鉴定等方法观察不同时期C57BL/6小鼠毛囊的发育情况。结果胚胎发育第15.5d(E15.5)开始出现毛囊,至E18.5胎鼠背部皮肤逐渐增厚,毛囊逐渐增多。乳鼠出生第1d毛囊大部分处于第4阶段之前,毛囊数量继续增加,至出生第4d皮肤基底层未见新生的毛囊,此时毛囊数量维持不变,出生第9d毛囊达到第8阶段。成鼠脱毛后第9d约有95%的毛囊进入生长Ⅳ期,第17d约有63.3%的毛囊进入退化期,第22d约有85%的毛囊处于退化期。结论小鼠毛囊的早期发育起始于E15.5,从E17.5到出生后3d毛囊数量呈快速增加趋势,因此,这一时期可以用于探讨毛囊再生机制等相关研究。     

汉坦病毒感染C57BL/6小鼠组织中特异性抗原的检测

目的通过检测汉坦病毒感染C57BL/6小鼠组织中特异性病毒抗原,以建立汉坦病毒感染动物的评价体系。方法将汉坦病毒陈株按照原病毒液、10-1、10-2三个滴度经肌肉注射感染C57BL/6小鼠,在感染后的第3、6、9、12、15天,分别取小鼠的心、肝、脾、肺、肾、脑等组织研磨后制成病毒悬液,以ELISA法检测各组织中的病毒特异性抗原。结果 C57BL/6小鼠感染汉坦病毒后短期内在其肝脏和脾脏可以检测到特异性抗原,随着时间的延长,这些抗原逐步消失。结论上述结果为建立汉坦病毒感染动物的评价体系提供了一种参考。     

Fatty acid bile acid conjugates inhibit atherosclerosis in the C57BL/6 mouse model.

OBJECTIVE: The aim of the current research was to study whether fatty acid bile acid conjugates (FABACs) have a beneficial effect on atherosclerosis progression and blood lipid levels in mice. METHODS: C57BL/6 female mice, fed a high-fat Paigen diet, were administered an oral dose of FABAC or DDH2O daily. Quantification of atherosclerotic fatty-streak lesions at the aortic sinus was performed. RESULTS: The FABAC-treated mice showed a significant reduction in the atherosclerotic lesion areas as compared to the control group (p = 0.019). A significant elevation in total cholesterol levels was observed in both the FABAC and control groups. Higher FABAC levels were measured in the high-density lipoprotein fraction as compared to the very-low-density and low-density lipoprotein fractions. CONCLUSION: Our findings demonstrate that FABACs, given orally, reduce the development of atherosclerosis in mice fed a high-fat high-cholesterol diet, despite a lack of effect on plasma lipid levels.     

A mammary gland adenomyoepithelioma in a C57BL/6 mouse

Mammary gland adenomyoepitheliomas are benign complex mammary gland tumors composed of neoplastic cells of epithelial and myoepithelial origins, described in many species (humans, dogs, cats, rats) and rarely in mice. We report here an adenomyoepithelioma in a C57BL/6 female mouse. Histologically, tubes and cords formed by neoplastic epithelial cells were separated by bundles of neoplastic myoepithelial cells in a clear and partially mucinous matrix. The tumor displayed characteristics of a benign neoplastic proliferation with a compressive growth pattern, and moderate cellular pleomorphism and mitotic index. At immunohistochemistry, the epithelial cells were strongly cytokeratin positive; the myoepithelial cells were weakly cytokeratin positive and strongly smooth muscle actin positive. This is to our knowledge, the first report of a mammary gland adenomyoepithelioma in a C57BL/6 mouse.     

A multimodal, multidimensional atlas of the C57BL/6J mouse brain

Strains of mice, through breeding or the disruption of normal genetic pathways, are widely used to model human diseases. Atlases are an invaluable aid in understanding the impact of such manipulations by providing a standard for comparison. We have developed a digital atlas of the adult C57BL/6J mouse brain as a comprehensive framework for storing and accessing the myriad types of information about the mouse brain. Our implementation was constructed using several different imaging techniques: magnetic resonance microscopy, blockface imaging, classical histology and immunohistochemistry. Along with raw and annotated images, it contains database management systems and a set of tools for comparing information from different techniques. The framework allows facile correlation of results from different animals, investigators or laboratories by establishing a canonical representation of the mouse brain and providing the tools for the insertion of independent data into the same space as the atlas. This tool will aid in managing the increasingly complex and voluminous amounts of information about the mammalian brain. It provides a framework that encompasses genetic information in the context of anatomical imaging and holds tremendous promise for producing new insights into the relationship between genotype and phenotype. We describe a suite of tools that enables the independent entry of other types of data, facile retrieval of information and straightforward display of images. Thus, the atlas becomes a framework for managing complex genetic and epigenetic information about the mouse brain. The atlas and associated tools may be accessed at http://www.loni.ucla.edu/MAP.     

The C57BL/6 nu/nu lpr/lpr mouse. I. Expression of the 'lpr phenotype' in the C57BL/6 genetic background

The C57BL/6 lpr/lpr mice develop within 3-6 moths a series of abnormalities of their immune system which characterize the 'lpr phenotype' and allow them to be distinguished easily from normal C57BL/6 as well as from C57BL/6 nu/nu: lymphadenopathy, hyperimmunoglobulinemia, high anti-nuclear antibody titers and, less regularly, anti-whole DNA antibody. Though C57BL/6 nu/nu occasionally present auto-antibody too, the combined use of three or four of the aforementioned parameters for each tested animal allows an easy detection of animals presenting the lpr phenotype. In our C57BL/6 lpr/lpr colony, the lymphadenopathy does not seem to start by chance in any lymphoid organ but shows a very strong preferential occurrence in the cervical lymph nodes. These parameters of the lpr phenotype have been used to trace the lpr genes and to construct the C57BL/6 nu/nu lpr/lpr as described in the companion paper.     

Standard atlas space for C57BL/6J neonatal mouse brain

A standard atlas space with stereotaxic co-ordinates for the postnatal day 0 (P0) C57BL/6J mouse brain was constructed from the average of eight individual co-registered MR image volumes. Accuracy of registration and morphometric variations in structures between subjects were analyzed statistically. We also applied this atlas coordinate system to data acquired using different imaging protocols as well as to a high-resolution histological atlas obtained from separate animals. Mapping accuracy in the atlas space was examined to determine the applicability of this atlas framework. The results show that the atlas space defined here provides a stable framework for image registration for P0 normal mouse brains. With an appropriate feature-based co-registration strategy, the probability atlas can also provide an accurate anatomical map for images acquired using invasive imaging methods. The atlas templates and the probability map of the anatomical labels are available at .     

Spontaneous central apneas occur in the C57BL/6J mouse strain

Despite the clinical significance of central apneas in a wide range of disorders little is known about their pathogenesis. Research in this field has been hindered by the lack of appropriate animal models. Our goal was to determine whether the C57BL/6J mouse strain, which has an inherited predisposition for dysrhythmic breathing, exhibits spontaneous apneas. In vivo plethysmography of unanesthetized, unrestrained adult C57BL/6J mice revealed a regular occurrence of spontaneous apneas. In situ recordings from respiratory outputs (phrenic, vagal, hypoglossal nerves) in the working heart-brainstem preparation (WHBP) also showed spontaneous central apneas accompanied by laryngeal closure as indicated by tonic vagal postinspiratory activity and increase in subglottal pressure. The apneas were further characterized by a hypoglossal discharge with delayed onset compared to the tonic vagal postinspiratory activity. We conclude that spontaneous central apneas with active laryngeal closure occur in C57BL/6J mice. This mouse strain is a useful animal model to study neuronal mechanisms that underlie the generation of spontaneous central apneas.