LP小鼠突变基因的定位及鉴定

目的以卷尾突变C57BL/6J小鼠为研究对象,通过微卫星定位以及候选基因测序分析确定突变基因位点。方法将LP小鼠与正常C57BL/6J及C3H小鼠交配,记录了后代中卷尾与正常小鼠的数目,确定卷尾突变表型的遗传模式。微卫星D1Mit113和D1Mit149对突变基因进行了精确定位,确定候选基因。PCR扩增候选基因片段直接测序并进行序列分析。用Fsp BI(Bfa I)内切酶鉴定LP小鼠杂交后代的基因型。结果 LP突变呈单基因不完全显性遗传,在不同的遗传背景中存在表型差异;Vangl2基因编码区1345bp处碱基由C→T。结论 Vangl2基因C→T的突变是一种无义突变,导致蛋白编码提前终止,是引起卷尾突变的原因;杂交LP小鼠后代中未出现纯合子小鼠,证明Vangl2基因突变纯合子致死。     

三种小鼠外周血淋巴细胞亚群正常值比较

目的测定健康正常Balb/C、C3H/HeJ和C3H/SW小鼠外周血淋巴细胞亚群含量，比较3种小鼠的差异性。方法用流式细胞仪测定Balb/C、C3H/HeJ和C3H/SW小鼠外周血淋巴细胞亚群含量。结果各20例3种小鼠、5种外周血淋巴细胞亚群含量呈正态分布，95%可信区间可以被认可；Balb/C与C3H/SW小鼠除B细胞外其他4种淋巴细胞亚群（T细胞、Ts细胞、Th细胞、NK细胞）含量差异性不大（P〉0.05）；而C3H/HeJ小鼠5种淋巴细胞亚群（T细胞、B细胞、Ts细胞、Th细胞、NK细胞）含量均比C3H/HeJ、C3H/SW小鼠低，并具有统计学意义（P〈0.05）。结论建立了健康正常Balb/C、C3H/HeJ和C3H/SW小鼠5种外周血淋巴细胞亚群含量正常参考值；C3H/HeJ小鼠细胞免疫功能比Balb/C、C3H/SW小鼠低，易受细菌等侵染。     

小鼠脊髓发育候选基因Wnt7b的初步研究

目的:探讨Wnt7b基因及Wnt信号系统分子对小鼠脊髓发育的作用。方法:基因测序比较A/J和C57BL/6J两种小鼠Wnt7b基因开放阅读框架序列差异;以RT-PCR方法测定Wnt7b基因在两种小鼠组织中的表达差异;免疫组化法测定Frizzled在2种孕14天胎鼠脊髓上的表达和分布。结果:测序结果中发现两种小鼠Wnt7b开放阅读框架序列结构具有差异;在2种小鼠8种组织中,脊髓、肌肉、脾和肾中的Wnt7b基因表达具有明显差异;免疫组化的测定表明,发育14天的胎鼠脊髓上的Frizzled表达强度和分布范围具有显著差异,尤其是A/J小鼠在神经管周围表达明显强于C57BL/6J。结论:基因表达差异和基因结构差异提示,Wnt7b基因与脊髓发育具有一定的关联,应为一脊髓发育候选基因;Frizzled的强表达与脊髓发育的抑制相关,Wnt7b的信号转导途径介导了控制脊髓发育的过程。     

近交系小鼠不同供受体组合对移植心脏存活期的影响

目的:在小鼠异位心脏移植模型中观察近交系小鼠不同品系的供受体组合对于移植心脏生存时间的影响.方法:采用腹腔异位小鼠心脏移植模型,术后通过扪诊评估移植心功能,比较C3H/HeJ、Balb/c和C57BL/6J常见近交系小鼠六种不同供受体组合生存时间的差异,并进行组织病理评分.结果:C3H/HeJ、Balb/c和C57BL/6J六种不同供受体组合的受体心脏存活时间无统计学差异,中位生存时间均为7～8 d,而且组内生存时间差异小;移植心脏组织病理检查发现所有标本均表现为严重的排斥反应,各组间无明显差异.结论:C3H/HeJ、Balb/c和C57BL/6J不同供受体组合均能符合移植免疫研究的要求,但是综合考虑手术难度、术后检测、喂养以及感染等因素,C57BL/6J和C3H/HeJ适合作为供体小鼠,而Ba1b/c更适合作为受体小鼠.     

小鼠脊髓发育遗传参数的研究

目的：进行小鼠脊髓发育形态学数据测定，研究遗传参数。方法：取4种近交系小鼠，A／J；C57BL／6J；DBA／2J；129xl／SvJ以及A／J与C57BL／6J杂交的F1和F2代重组个体，多聚甲醛灌注后进行脊髓重量、颈膨大截面积、灰质百分比的测定。结果：数据显示在4个近交系中，A／J与C57BL／6J脊髓发育差别最大，A／J为小脊髓(平均重65．21mg，截面积2．69mm^2；C57BL／6J为大脊髓，平均重81．58mg，截面积3．06mm^2)。在该近交系与重组小鼠中，脊髓发育的遗传度为30％～40％，最少控制基因数为2～4。结论：小鼠中脊髓发育存在超显性现象，通过小鼠脊髓形态测定获得了发育相关的遗传参数，为脊髓发育数量性状基因座(QTL)和控制基因的确认提供了资料。     

小鼠脊髓腰膨大截面积QTL研究

目的 :小鼠脊髓腰膨大截面积性状进行数量性状基因座 (Quantitativetraitlocus ,QTL)的分析定位。方法 :应用近交系小鼠A/J和C5 7BL/6J以及其F1、F2代为材料 ,测定脊髓腰膨大截面积 ,利用分布于小鼠全基因组的 87对引物 ,PCR法进行F2代小鼠基因组扫描分析。结果 :9个QTL与脊髓腰膨大截面积相关 ,分别位于 4,8,13 ,14 ,15 ,16,17和X染色体上。其中 7个QTL的性状变异解释率 (10 % )。对照我们以前的结果 ,3个腰膨大QTL与脊髓重和颈膨大QTL均重叠 ;3个与脊髓重QTL重叠 ;3个单独与腰膨大相关。D15Mit15 8位点显示出最大的性状变异解释率 (15 % ) ,与脊髓重最大的性状变异解释率 (2 4% )的位点相吻合。结论 :研究结果说明控制小鼠脊髓发育最主要位点位于 15号染色体D15Mit15 8附近。本研究为小鼠脊髓发育QTL的精细定位、进行位置克隆以最终确认控制基因打下了基础     

EHEC：O157感染后小鼠肾细胞凋亡的检测

目的 检测C3H/HeJ和C3H/SW小鼠感染肠出血性大肠杆菌(EHEC:O157)后肾细胞凋亡,并探讨thl4基因突变对肾细胞凋亡的影响.方法 流式细胞术(FCM)检测C3H/HeJ和C3H/SW小鼠感染EHEC:O157后肾细胞凋亡率,采用SPSS11.0统计软件分析结果.结果 C3H/HeJ小鼠肾细胞凋亡率明显高于C3H/SW小鼠,差异有统计学意义(P<0.05).结论 小鼠感染EHEC:O157后肾细胞凋亡与thl4基因突变相关.     

小鼠脊髓腰膨大发育的初步遗传分析

目的 :筛选小鼠脊髓腰膨大发育具有差异的近交系。检测小鼠各世代群体中个体的数量性状值 ,研究遗传因素在小鼠腰膨大发育中贡献的大小。方法 :取 4种近交系小鼠 ,A/J;C5 7BL/6J ;DBA/2J ;12 9xl/SvJ以及A/J与C5 7BL/6J杂交获得的F1和F2个体 ,多聚甲醛灌注后进行腰膨大截面积测定。结果 :四个近交系中 ,A/J与C5 7BL/6J脊髓腰膨大截面积差别显著。脊髓腰膨大发育遗传度为 3 5 % ,最少控制基因对数为 2。结论 :小鼠脊髓腰膨大发育的数量性状值与遗传参数的确定 ,为其数量性状基因座的确定提供了有价值的资料。     

精神压力对C3H/HeJ小鼠斑秃发病率的影响

目的 研究精神压力对C3H/HeJ小鼠斑秃发病率的影响.方法 选取C3H/HeJ小鼠80只,随机分为4组,即精神压力组、药物诱导组、精神压力与药物诱导组和对照组,每组20只,干预后观察4个月,统计斑秃发病率,并比较各组差异.结果 精神压力组斑秃发病率为10%,对照组没有发生;精神压力与药物诱导组发病率为90%,药物诱导组发病率为60%,差异有统计学意义（P〈0.05）.结论 精神压力在药物诱导下显著提高C3H/HeJ小鼠斑秃发病率.     

TLR4在华支睾吸虫感染致小鼠肝纤维化作用的研究

目的探讨TLR4基因在华支睾吸虫感染小鼠致肝纤维化中的作用。方法建立感染华支睾吸虫C3H/HeN(TLR4野生型)与C3H/HeJ(TLR4基因突变型)小鼠模型,在感染后1d、7d、2W、4W、8W、12W取小鼠肝脏,切片并进行HE和Masson染色,观察病理变化。结果 C3H/HeN小鼠肝脏大体病变从2W开始,随感染时间延长而加重;C3H/HeJ小鼠从2W开始,4W加重,至8W已明显减轻。观察HE、Masson染色肝脏切片示随感染时间的延长,C3H/HeN小鼠前期肝纤维化逐渐加重,至8W起趋于稳定;2W~12W纤维化评分均高于感染前(P0.01)。C3H/HeJ小鼠肝纤维化评分2W升高,4W达高峰并显著高于感染前(P0.01),至8W明显下降。与C3H/HeN比较,感染后2W、4W、8W、12W C3H/HeJ小鼠肝纤维化程度轻(P0.01)。感染后7d,C3H/HeN小鼠的HE染色切片上可见嗜酸性粒细胞,4W达高峰,此后明显减少。感染后2W、4W、8W,C3H/HeJ小鼠嗜酸性粒细胞较C3H/HeN小鼠少(P0.05)。结论 TLR4基因缺失可能在华支睾吸虫感染致肝纤维化过程起一定的阻制作用,从而减缓肝纤维化的进程。     

Effects of endotoxin on carbohydrate metabolism in inbred mice.

Effects of endotoxin on carbohydrate metabolism were studied in A/HeJ (endotoxin-sensitive) and C3H/HeJ (endotoxin-resistant) inbred mice. A/HeJ mice developed hypoglycemia within two hours after endotoxin injection, yet liver glycogen content did not differ from controls. Similarly treated C3H/HeJ mice did not develop significant hypoglycemia. Administration of glucagon to endotoxin-treated A/HeJ mice failed to elevate their blood glucose concentrations, while endotoxin-treated mice of the same strain did respond to dibutyryl cyclic AMP with a significant elevation of blood glucose. C3H/HeJ mice on the other hand responded to glucagon and dibutyryl cyclic AMP with elevated blood glucose. Endotoxin-treated C3H/HeJ but not A/HeJ mice were able to carry out gluconeogenesis induced by prednisolone, while both inbred strains showed active glycogenesis after administration of an exogenous glucose load. Administration of glucagon resulted in diminished liver glycogen concentrations in A/HeJ endotoxin-treated mice suggesting no impairment of glycogenolysis. The inability of endotoxin-treated A/HeJ mice to respond to glucagon could be due to impairment of gluconeogenesis. Although endotoxin interfered with the capacity of both inbred strains to respond to glucagon administration with elevation of liver cyclic AMP, the effect was significantly more severe in A/HeJ mice. The susceptibility of A/HeJ mice to the lethal effect of endotoxin may be related to the apparent sensitivity of carbohydrate metabolic pathways to disturbance by endotoxin.     

The Collaborative Cross mouse genetic reference population designed for dissecting complex traits

Complex traits are multifactorial traits controlled by polygenic host factors. These trait-related phenotypic characteristics and performance including body weight, blood chemistry, immune cell profiles, as well host susceptibility to infectious and chronic diseases. In recent years, tremendous efforts were invested aiming to map the host genetic factors attribute to these traits and subsequently clone the gene/s underlying these loci. In parallel to human studies, a number of mouse models and approaches were developed aimed to enhance the mapping process and the gene cloning. These include of using resources such as F2, backcross, advanced intercross lines, outbred populations, consomic, congenic and recombinant inbred lines (RIL). The constraints of these approaches were the limited resolution mapping of genomic regions of the quantitative trait loci (QTL) associated with the trait of interests, and the limited genetic diversity observed in the parental founders. To overcome these limitations, a new genetically highly diverse recombinant inbred lines of mouse population was established, namely the Collaborative Cross (CC), created from full reciprocal mating of 8 divergent strains of mice:A/J, C57BL/6J, 129S1/SvImJ, NOD/LtJ, NZO/HiLtJ, CAST/Ei, PWK/PhJ, and WSB/EiJ. By intercrossing these eight founders to generate the different CC lines, the genetic makeup of the newly developed resource is completely different from the eight parental lines, and will show heterosis, which subsequently will response differently comparing with their original founders. Finally, our results suggest that it is not essential to defining the phenotypic response of the eight parental lines, prior of assessing the CC lines, because it is believed that genetic interaction of the new genetic makeup of the new lines will reveal new phenotypic response, which completely different from the parental lines.In this report, we present to the community the power of the CC for dissecting variety of complex traits including host susceptibility to infectious and chronic diseases as well body performance traits. Based on our results from a variety of studies, we recommend to the community, that the best strategy of using this population is to aim of phenotyping about 50 and more of CC lines, with limited number of biological replicates (3-4 mice per line), and subsequently using the publicly available high dense genotype information of the CC lines as well the sequence database of the eight founders, it will be possible performing QTL mapping to a unprecedented precision genomic regions less than 1 MB, subsequently lead to identify potential strong candidate genes. These achievements are believed cannot be obtained with any other currently available mouse resource populations.     

C57BL/6J和DBA/2J小鼠可能与学习记忆相关基因的初步筛选

目的：初步筛选出C57BL/6J（B6）和DBA/2J（D2）小鼠海马中可能与学习记忆有关的新基因，为以后的学习记忆基因研究提供参考。方法：利用已完成基因组测序的 B6、D2两个近交系小鼠的海马全基因（45101个）表达的数据，采用差异检验、功能分析方法进行筛选。结果：共筛选出5个可能与学习记忆有关的候选基因：cntnap2、pacsin1、cntn1、lypd1、ndrg2。结论：候选基因可能与已知的学习记忆机制的组成部分有关，增强了筛选的预见性和科学性，并将进一步确认候选基因与学习记忆的关系。     

克雷伯氏肺炎杆菌内毒素诱导小鼠β—防御素—4基因表达及其信号传递

目的 探讨用克雷伯氏肺炎杆菌内毒素（LPS）对小鼠β－防御素表达的影响及其相关的信号传导通路。方法 分别给予C3H／HeJ和C3H／HeN小鼠腹腔注射LPS（4mg/kg),于不同时间点采取气管、肺、肾管组织,提取总RNA。用RT－PCR检测各组织β-防御素－3和／或β-防御素－4mRNA的表达,并对扩增出的cDNA片段进行测序;同步用Western blot法检测该两系小鼠肺脏1－kBa磷酸化情况（p-I-kBa)和I－kBa的量。结果 （1）经LPS处理24小时后的C3H／HeN小鼠,其肺脏表达β-防御素－4mRNA而C3H／HeJ小鼠未见表达。（2）与未给予LPS处理小鼠比较,经LPS处理4小时后,C3H／HeN小鼠肺组织p-kBa含量明显增高;LPS处理后8小时,p-I-kBa以及I-kBa含量均呈减少趋势;至第24小时,p-I-kBa和I－kBa含量均明显减少。而C3H／HeJ小鼠肺组织p-I-kBa及I－kBa含量均未见相应变化。结论 克雷伯氏肺炎杆菌内毒素能诱导C3H／HeN小鼠肺β-防御素－4的表达,其诱导表达作用与Toll受体蛋白－4介导的NF－kB活化级联信号传递通路有关。     

6个近交系小鼠线粒体DNA D-Loop区PCR-RFLP分析

分析Ｔ７３９等６个近交系小鼠线粒体ＤＮＡＤ－Ｌｏｏｐ区的多态性,探讨近交系小鼠的遗传监测方法。方法：ＰＣＲ－ＲＦＬＰ技术,即ＰＣＲ技术结合限制性片段长度多态分析。结果Ｄ－Ｌｏｏｐ片段经ＨａｅⅢ、ＨｉｎｆⅠ、ＥｃｏＲⅤ、ＩＮＤⅢ、ＨｐａⅠ、ＢａｍＨⅠ、ＡｐａⅠ、ＮｄｅⅡ、ＸｈｏⅠ、ＸｂａⅠ内切酶消化后,Ｔ７３９、ＴＡ２、６１５、ＢＡＬＢ／Ｃ、Ｃ３Ｈ、Ｃ５７ＢＬ／６Ｊ小鼠表现出完全相同的酶切栈局,未发     

克雷伯氏肺炎杆菌内毒素诱导小鼠β-防御素-4基因表达及其信号传递

目的　探讨用克雷伯氏肺炎杆菌内毒素 (L PS)对小鼠 β-防御素表达的影响及其相关的信号传导通路。方法　分别给予 C3H /He J和 C3H /He N小鼠腹腔注射 L PS (4mg/kg) ,于不同时间点采取气管、肺、肾等组织 ,提取总 RNA。用 RT- PCR检测各组织β-防御素 - 3和 /或β-防御素 - 4m RNA的表达 ,并对扩增出的 c DNA片段进行测序 ;同步用 Western blot法检测该两系小鼠肺脏 I-κBα磷酸化情况 (p- I-κBα)和 I-κBα的含量。结果　 1经L PS处理 2 4小时后的 C3H/He N小鼠 ,其肺脏表达 β-防御素 - 4m RNA 而 C3H/He J小鼠未见表达 ;2与未给予L PS处理小鼠比较 ,经 L PS处理 4小时后 ,C3H/He N小鼠肺组织 p- I- κBα含量明显增高 ;L PS处理后 8小时 ,p- I-κBα以及 I- κBα含量均呈减少趋势 ;至第 2 4小时 ,p- I- κBα和 I- κBα含量均明显减少。而 C3H/He J小鼠肺组织 p- I-κBα及 I- κBα含量均未见相应变化。结论　克雷伯氏肺炎杆菌内毒素能诱导 C3H/He N小鼠肺 β-防御素 - 4的表达 ;其诱导表达作用与 Toll受体蛋白 - 4介导的 NF-κB活化级联信号传递通路有关