C57BL/6J-HBV转基因小鼠的繁育与检测

目的　摸清C57BL 6J -HBV转基因小鼠的繁育规律 ,建立可靠的HBsAg表达检测方法。方法　对C57BL 6J-HBV转基因小鼠两种不同交配方式的繁殖性能和HBsAg的表达情况进行比较研究 ;应用免疫组化的方法证实血清学诊断HBsAg的准确性并用激光扫描共聚焦显微镜确定HBsAg在肝细胞中的表达部位。结果与结论　从繁殖性能来看 ,两种交配方式窝产仔数差异不显著 (P >0 .0 5 ) ,而离乳数差异具有显著性 ( 0 .0 1

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C57BL/6小鼠性别差异对髓鞘少突胶质糖蛋白诱导的实验性自身脑脊髓炎疾病的影响

多发性硬化是人类常见的中枢神经系统自身免疫性炎症致脱髓鞘疾病．流行病学研究发现,女性患者多于男性,其平均发病时间早于男性．实验性自身免疫性脑脊髓炎(EAE)与多发性硬化症有相似的临床症状和病理特征,是被广泛应用于人类疾病研究的动物模型．本实验利用髓鞘少突胶质糖蛋白MOG_33-35免疫C57BL/6小鼠建立EAE模型,观察29天．通过疾病评分发现雌雄小鼠在发病率、起病时间上均无明显差别,但雄鼠的发病症状明显比雌鼠严重．在其病理切片HE染色中观察到雄性小鼠中枢浸润的炎性细胞多于雌性小鼠,并且在LFB染色中同样观察到雄鼠脱髓鞘区域明显增大．对其发病高峰期中枢浸润细胞的染色分析时,可以发现雄性小鼠中浸润的CD4 T细胞及其亚群T_H-1和T_H-17细胞均有明显增加．这些都表明MOG_33-35免疫C57BL/6小鼠建立的EAE模型存在着性别差异的影响,这一发现为今后建立多发性硬化症的动物模型中动物性别的选择提供了一定的参考依据．     

不同方法建立C57BL/6J小鼠抑郁症模型的探讨

目的用不同方法建立C57BL/6J小鼠抑郁症模型,为探讨GalR蛋白对小鼠抑郁症的治疗作用打下基础。方法 C57BL/6J小鼠经体重、敞箱实验及反抗抓获实验初筛后,随机分为4组:Ⅰ.CUMS组,Ⅱ.CUMS+CORT组,Ⅲ.CORT组,Ⅳ.正常对照组。每天记录小鼠体重及摄食量。28d后进行液体消耗及强迫游泳实验测试。结果小鼠抑郁模型在第28d建立成功。Ⅱ组小鼠短期内体重迅速下降并死亡。与Ⅰ组小鼠相比,Ⅲ组小鼠液体消耗和强迫游泳实验指标改变更明显。结论成功建立C57BL/6J小鼠抑郁症模型。CUMS和CORT模型结合,小鼠不能耐受,短期内死亡。单独CORT模型造模效果要优于CUMS模型。在后续试验中,将用CORT法建立C57BL/6J小鼠抑郁症模型。     

Genomic Imprinting and Problem of Parthenogenesis in Mammals

Genomic imprinting belongs by its nature to problems of epigenetics, which studies hereditary changes in gene expression not related to defective sequences of DNA nucleotides. Epigenetic mechanisms of control, including genomic imprinting, are involved in many processes of normal and pathological development of humans and animals. Disturbances of genomic imprinting may lead to various consequences, such as formation of developmental anomalies and syndromes in humans, appearance of the large offspring syndrome and increased mortality upon cloning of mammals, and death of parthenogenetic embryos soon after implantation and beginning of organogenesis. The death of diploid parthenogenetic or androgenetic mammalian embryos is determined by the absence of expression of the genes of imprinted loci of the maternal or paternal genome, which leads to significant defects in development of tissues and organs. A review is provided of the studies aimed at search of possible normalization of misbalanced gene activity and modulation of genomic imprinting effects during parthenogenetic development in mammals.__________Translated from Ontogenez, Vol. 36, No. 4, 2005, pp. 300–309.Original Russian Text Copyright © 2005 by Platonov.     

C57BL／6J-HBV转基因小鼠血清生化指标与性别年龄的关系

目的观察C57BL/6J-HBV乙型肝炎病毒转基因小鼠血清总胆红素（T-BIL）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总蛋白（TP）和白蛋白（ALB）与性别和年龄的关系,以及与遗传背景相同的C57BL／6J小鼠的差异。方法选取8周龄和24周龄的C57BL／6J-HBV转基因小鼠及C57BL／6J小鼠的血清测定T-BIL、ALT、AST、TP和ALB值。结果C57BL／6J-HBV转基因小鼠与同周龄同性别的C57BL／6J小鼠相比,T-BIL、ALT、AST、TP和ALB均存在显著差异（P〈0．05）;24周龄的C57BL／6J-HBV转基因小鼠ALT和AST与其8周龄鼠相比均存在显著差异（P〈0．05）。结论C57BL／6J-HBV转基因小鼠T-BIL、ALT、AST、TP和ALB值显著高于C57BL／6J小鼠;且C57BL／6J-HBV转基因小鼠ALT和AST值与年龄有关,与性别无关。     

Blueberry and Mulberry Juice Prevent Obesity Development in C57BL/6 Mice

Objectives

To establish whether blueberry (Vaccinium ashei) and mulberry (Morus australis Poir) juice, anthocyanin rich fruit juice, may help counteract obesity.

Design

And Methods: Four-week-old C57BL/6 mice were fed a high-fat diet (HFD) with or without blueberry and mulberry juice for 12 weeks. Body weight, serum and hepatic lipids, liver and adipose tissues morphology, insulin and leptin were assessed.

Results

Mice fed HFD exhibited increased body weight, insulin resistance, serum and hepatic lipids. In comparison, blueberry and mulberry juice inhibited body weight gain, decreased the serum cholesterol, reduced the resistance to insulin, attenuated lipid accumulation and decreased the leptin secretin.

Conclusion

These results indicate that blueberry and mulberry juice may help counteract obesity.     

链脲佐菌素诱导C57BL/6J小鼠2型糖尿病模型研究

目的建立与2型糖尿病(非胰岛素依赖型糠尿病、NIDDM)病人临床特征和发病过程相似的NIDDM动物模型.方法用高脂肪饲料喂养C57BL/6J雄性断乳小鼠3周,腹腔注射链脲佐菌素(STZ),继续喂养4周,测定给药前和给药后1、3、4周非空腹血糖、实验结束时非空腹胰岛素水平,观察胰腺形态学变化.结果喂养3周后(给药前)高脂饲料-STZ组及高脂饲料-柠檬酸组血糖浓度(7.0±0.39)mmol/L、( 6.8±0.45)mmol/L高于普通饲料-STZ组及普通饲料-柠檬酸组(5.3±0.40)mmol/L、(5. 4±0.39)mmol/L,P＜0.05;实验结束时,高脂饲料-STZ组血糖浓度(13 .1±2.01)mmo/L高于高脂饲料-柠檬酸(6.9±0.46)mmol/L、普通饲料-柠檬酸组(6.0± 0.46)mmol/L和普通饲料-STZ组(7.1±0.62)mmol/L(P＜0.05),各组间血浆胰岛素浓度、体重及饮水量差异无显著性,P＞0.05;实验过程中高脂饲料STZ组和柠檬酸组小鼠每天进食热量(64.49±9.2)kJ/只,(70.7±9.6)kJ/只, 显著高于普通饲料STZ组和柠檬酸组(52.7±7.9)kJ/只,(57.3±11.7)kJ/只;各组小鼠胰腺和胰岛细胞形态正常.结论高脂肪饲料和STZ是用C57BL/6J断乳幼鼠建立NIDDM模型所必须的,100mg/kg体重STZ对普通饲料小鼠血糖无影响;用高脂饲料和STZ 处理的小鼠血糖升高、胰岛素浓度正常,与NIDM病人临床特征和发病过程相似;C57BL/6J小鼠易得,建模方法简便,费用低,是在NIDDM实验研究中能广泛使用的较理想的非遗传性NID DM动物模型.     

Different influences of genomic imprinting on the development of parthenogenetic cell clones in C57BL/6 and CBA mice

Clonal analysis of parthenogenetic chimeric mouse embryos C57B1/6(PG)<-->BALB/c has shown that parthenogenetic cell clones C57BL/6 are present in the brain, liver, and kidneys of 14- and 18-day-old embryos. The content of the parthenogenetic component (PG) in these organs on day 18 was lower than on day 14, and, in some 18-day-old embryos, parthenogenetic cell clones were absent from the liver and/or kidneys. These data suggest that, during the embryogenesis of parthenogenetic chimeras, parthenogenetic cell clones of mostly endodermal and mesodermal origins were actively eliminated. Therefore, in such parthenogenetic adult chimeras, parthenogenetic clones of mostly ectodermal origins were preserved. In parthenogenetic chimeras CBA(PG)<-->BALB/c, parthenogenetic cell clones were actively eliminated at early embryonic stages, and, as a result, they were absent at the post-implantation stages. Hence, during development of parthenogenetic cell clones, the effects of genomic imprinting are expressed unequally in C57BL/6 and CBA mice.     

C57BL/6小鼠听皮层A1区LTP特性

应用在体细胞外记录群体细胞兴奋性突触后场电位方法,研究C57BL6小鼠听皮层A1区突触长时程增强(LTP)特性。观察到,给予模拟的θ节律电刺激(TBS),刺激听皮层白质,可在听皮层灰质ⅡⅢ层记录到明显LTP。根据TBS后LTP幅度变化特征,LTP可分为瞬时增强型(有PTP)和缓慢增强型。高强度TBS诱导的LTP增幅百分数比低强度TBS诱导的LTP增幅大,但两者诱导的LTP成功率无显著差异。实验还表明,2月龄小鼠较4月龄小鼠LTP幅度增长率更高,提示听皮层LTP具有年龄依赖性特征。实验结果为进一步研究听觉模态学习记忆的神经机制提供了资料。     

Garcinia cambogia Extract Ameliorates Visceral Adiposity in C57BL/6J Mice Fed on a High-Fat Diet

The aim of present study is to evaluate the effects of Garcinia cambogia on the mRNA levels of the various genes involved in adipogenesis, as well as on body weight gain, visceral fat accumulation, and other biochemical markers of obesity in obesity-prone C57BL/6J mice. Consumption of the Garcinia cambogia extract effectively lowered the body weight gain, visceral fat accumulation, blood and hepatic lipid concentrations, and plasma insulin and leptin levels in a high-fat diet (HFD)-induced obesity mouse model. The Garcinia cambogia extract reversed the HFD-induced changes in the expression pattern of such epididymal adipose tissue genes as adipocyte protein aP2 (aP2), sterol regulatory element-binding factor 1c (SREBP1c), peroxisome proliferator-activated receptor γ2 (PPARγ2), and CCAT/enhancer-binding protein α (C/EBPα). These findings suggest that the Garcinia cambogia extract ameliorated HFD-induced obesity, probably by modulating multiple genes associated with adipogenesis, such as aP2, SREBP1c, PPARγ2, and C/EBPα in the visceral fat tissue of mice.     

Immunological,Cytogenetical, and Behavioral Changes in CBA and C57BL/6 Male Mice after Pheromonal Action

Immunological and cytogenetical changes were studied in mice-recipients of CBA and C57BL/6 lines as well as modifications of their behavior appearing as a result of action of various stressors. Post-stress volatile excretions of CBA males cause led to a pronounced immunosuppression in males-recipients of both lines. This resulted in a decrease of capability for immune response of the stressed mice to sheep erythrocytes. Besides, in young males of the CBA line, frequency of chromosome aberrations in bone marrow cells increased statistically significantly after stressing by syngenec volatile excretions of mature animals. Attractive properties of volatile excretions of the CBA and C57BL/6 line males were studied. In norm, animals of both lines prefer syngenic odors of intact males donors. However, for the CBA line males, post-stress excretions from males-donors of both lines became more attractive than excretions from intact animals. Also revealed were interline differences in incidence of choice of syngenic and allogenic post-stress excretions. This can be explained both by different stress-reactivity of animals-donors and differences of recipients of the studied genotypes in selectivity to olfactory stimuli. An importance of olfactory stresses and of the revealed interline differences for explanation of some micro-and macroevolutionary synecological processes in the domestic mouse is discussed.     

FMR1基因敲除对C57BL/6小鼠部分生物学特性的影响

为探讨脆性X智力障碍1(FMR1)基因敲除对C57BL/6小鼠(Mus musculus domesticus)部分生物学特性的影响,使用全自动动物血细胞分析仪和全自动生化仪分别检测8~10周龄的FMR1基因敲除小鼠(C57BL/6 FMR1 KO)及同源背景C57BL/6小鼠的血液生理生化指标、电解质等,并进行统计学分析。C57BL/6 FMR1 KO小鼠与野生型C57BL/6小鼠比较,血液生理指标中雌性及雄性组间的中性粒细胞计数(MEUT#)、中性粒细胞百分比(MEUT)和淋巴细胞百分比(LY)存在显著性差异(P0.05);雄性组间的红细胞总数(RBC)、血红蛋白(HGB)、红细胞压积(HCT)和平均血红蛋白浓度(MCHC)存在显著性差异(P0.05);雌性组间的白细胞(WBC)、淋巴细胞计数(LY#)存在显著性差异(P0.05);而非性别因素影响两类小鼠组间的红细胞总数(RBC)、红细胞压积(HCT)、中性粒细胞计数(MEUT#)、中性粒细胞百分比(MEUT)和淋巴细胞百分比(LY)存在显著性差异(P0.05);血清生化指标中雄性组间的谷草转氨酶/谷丙转氨酶(AS/AL)存在显著性差异(P0.05);雌性组间的肌酐CR、肌酸磷酸激酶CK和钙离子浓度Ca2+存在显著性差异(P0.05);而非性别因素影响两类小鼠组间的谷草转氨酶/谷丙转氨酶(AS/AL)、肌酐CR和肌酸磷酸激酶CK存在显著性差异(P0.05);其他各项指标差异不显著(P0.05)。研究表明,FMR1基因敲除可影响小鼠的部分生理生化指标水平,这为今后研究和应用C57BL/6 FMR1 KO小鼠模型提供了实验依据。     

Effect of chronic corticosterone application on depression‐like behavior in C57BL/6N and C57BL/6J mice

Many studies using genetic mouse models are performed with animals on either one of the two closely related genetic backgrounds, C57BL/6J or C57BL/6N. These strains differ only in a few genetic loci, but have some phenotypic differences that also affect behavior. In order to determine the effects of chronic stress hormone exposure, which is relevant for the pathogenesis of psychiatric disorders, we investigated here the behavioral manifestations of long‐term increase in corticosterone levels. Thus, male mice from both sub‐strains were subcutaneously implanted with corticosterone (20 mg) or placebo pellets that released the hormone for a period of 21 days and resulted in significantly elevated plasma corticosterone levels. Corticosterone significantly increased food intake in B6N, but not in B6J mice. At various time points after pellet implantation, we performed tests relevant to activity and emotional behaviors. B6J mice displayed a generally higher activity in the home cage and the open field. Corticosterone decreased the activity. In B6N mice, corticosterone also decreased sucrose preference, worsened the coat state and increased forced swim immobility, while it had no effect in the B6J strain. Altogether, these results indicate that B6N mice are more sensitive to some of the effects of chronic corticosterone treatment than B6J mice.     

Increased Intestinal Epithelial Cell Turnover and Intestinal Motility in Gymnophalloides seoi-Infected C57BL/6 Mice

The changing patterns of goblet cell hyperplasia, intestinal epithelial cell turnover, and intestinal motility were studied in ICR and C57BL/6 mice infected with Gymnophalloides seoi (Digenea: Gymnophallidae). Whereas ICR mice retained G. seoi worms until day 7 post-infection (PI), C57BL/6 mice showed a rapid worm expulsion within day 3 PI. Immunosuppression with Depo-Medrol significantly delayed the worm expulsion in C57BL/6 mice. Goblet cell counts were increased in both strains of mice, peaking at day 1 PI in C57BL/6 mice and slowly increasing until day 7 PI in ICR mice. In C57BL/6 mice infected with G. seoi, newly proliferating intestinal epithelial cells were remarkably increased in the crypt, and the increase was the highest at day 1 PI. However, in ICR mice, newly proliferating intestinal epithelial cells increased slowly from day 1 to day 7 PI. Intestinal motility was increased in G. seoi-infected mice, and its chronological pattern was highly correlated with the worm load in both strains of mice. Meanwhile, immunosuppression of C57BL/6 mice abrogated the goblet cell proliferation, reduced the epithelial cell proliferation, and suppressed the intestinal motility. Goblet cell hyperplasia, increased intestinal epithelial cell turnover, and increased intestinal motility should be important mucosal defense mechanisms in G. seoi-infected C57BL/6 mice.     

Perinatal Exposure to Low-Dose Methoxychlor Impairs Testicular Development in C57BL/6 Mice

Methoxychlor (MXC), an organochlorine pesticide, has adverse effects on male reproduction at toxicological doses. Humans and wild animals are exposed to MXC mostly through contaminated dietary intake. Higher concentrations of MXC have been found in human milk, raising the demand for the risk assessment of offspring after maternal exposure to low doses of MXC. In this study, pregnant mice (F0) were given intraperitoneal daily evening injections of 1 mg/kg/d MXC during their gestational (embryonic day 0.5, E0.5) and lactational periods (postnatal day 21.5, P21.5), and the F1 males were assessed. F1 testes were collected at P0.5, P21.5 and P45.5. Maternal exposure to MXC disturbed the testicular development. Serum testosterone levels decreased, whereas estradiol levels increased. To understand the molecular mechanisms of exposure to MXC in male reproduction, the F1 testes were examined for changes in the expression of steroidogenesis- and spermatogenesis- related genes. RT-PCR analysis demonstrated that MXC significantly decreased Cyp11a1 and increased Cyp19a1; furthermore, it downregulated certain spermatogenic genes (Dazl, Boll, Rarg, Stra8 and Cyclin-a1). In summary, perinatal exposure to low-dose MXC disturbs the testicular development in mice. This animal study of exposure to low-dose MXC in F1 males suggests similar dysfunctional effects on male reproduction in humans.     

Utility of a C57BL/6 ES line versus 129 ES lines for Targeted Mutations in Mice

Inbred ES lines, though useful for generating targeted mutations in mice, are used infrequently. To appreciate the relative efficiency of inbred ES lines, a C57BL/6 ES line was compared with 129 strain ES lines for effectiveness in chimera formation leading to the establishment of targeted mutations in mice. Data from a transgenic facility spanning 7 years were collected. C57BL/6 ES cells injected into Balb/c embryos results in lower coat color chimerism than do 129 ES cells injected into C57BL/6 embryos. Combined data indicate that five independent targeted C57BL/6 clones should be injected as compared to three independent 129 clones to generate enough chimeras to effectively test for germ-line transmission. Thus, although less efficient than 129 ES lines, the C57BL/6 ES line is a relatively competent line and useful for the routine generation of targeted mutations in mice on a defined genetic background.     

Effects of fibroblast growth factor 2 and insulin-like growth factor II on the development of parthenogenetic mouse embryos in vitro

Most parthenogenetic embryos (PEs) in mammals die shortly after implantation, and this failure to develop is associated with genomic imprinting. We have examined the influence of human recombinant basic fibroblast growth factor 2 (FGF-2) and human recombinant insulin-like growth factor II (ICF-II) on the development of (CBA x C57BL/6)F1 parthenogenetic mouse embryos. Embryos were treated in vitro at the morula stage with different doses of FGF-2 and, after their development to blastocysts, transferred to pseudopregnant recipients. The optimal doses of FGF-2 did not affect the number of forming and implanting blastocysts, but increased, from 20 to 42%, the number of embryos developing to somite stages. PEs (18-21 somites) treated with an optimal dose of FGF-2 were explanted for further development in culture by treatment with the second growth factor, IGF-II. Eighty-three percent of those embryos cultured with IGF-II (2.5 microg/ml) developed to 35 or more somites, as compared with 36% of embryos cultured without any growth factors (P < 0.01). Also, a significantly higher proportion of PEs developed to 40-50 somites in this case. These results show that the in vitro treatment of PEs with FGF-2 at the morula stage increases the number of somite embryos, and the second treatment of somite PEs with IGF-II in culture medium prolongs their development significantly.     

Immune responses against allogeneic and syngeneic tumors in aged C57BL/6 mice

Aged C57BL/6 (B6) mice could reject allogeneic BALB/c RL male 1 tumor as efficiently as young B6 mice. However, in vitro analysis showed impaired generation of cytotoxic T cell response in aged B6 mice against allogeneic tumor. The reaction could be augmented by the addition of recombinant interleukin-2 (rIL-2). Enzyme-linked immunospots (ELISPOT) produced by CD8+ T cells purified from spleen cells showed no reduction in aged mice. The findings suggested that the number of CD8+ T cells capable of reacting against allogeneic H-2 antigens was similar in young and aged B6 mice. Low cytotoxic T lymphocyte (CTL) responsiveness in aged B6 mice appeared to have resulted from low responsiveness of CD4+ T cells producing IL-2. Although CTL generation was apparently impaired, strong multiple antigenicity of allogeneic tumor evoked a rejection response in aged B6 mice. On the other hand, no rejection response was observed against syngeneic EL4 tumor in aged B6 mice even after depletion of CD4+ CD25+ immunoregulatory cells. Depletion of CD4+ CD25+ cells caused rejection of EL4 tumor in young B6 mice. The findings suggested that aged B6 mice were incapable of inducing effector cells against weak tumor antigens. Only marginal CTL response and small number of ELISPOTs were generated in young but not aged B6 mice against EL4. Addition of rIL-2 to the culture augmented EL4 killing and ELISPOTs in spleen cells from young and aged B6 mice.     

从C57BL/6J小鼠胚胎中分离ES细胞系(英文)

ＥＳ细胞（ＥｍｂｒｙｏｎｉｃＳｔｅｍＣｅｌｓ）是来源于小鼠早期胚胎的细胞系,它可以在体外大量培养而不失去其发育的多潜能性。ＥＳ细胞不仅可以用来制作转基因动物,而且能够作为载体进行基因打靶等多种研究。目前,国际上常用的胚胎干细胞系都是来源于１２９小鼠的胚胎。因而,有必要探讨用其它品系小鼠建立ＥＳ系。１９９１年,Ｌｅｄｅｒｍａｎｎ等人首次报道从Ｃ５７ＢＬ／６Ｊ小鼠胚胎中建成了ＥＳ细胞系,但是没有对建立的细胞系进行特性分析。国内柴桂萱等人虽然做了特性分析,但是他们所建细胞系的细胞直径较大,生长速度较慢,不同于常见的ＥＳ细胞系。我们从Ｃ５７ＢＬ／６Ｊ品系小鼠胚胎中共分离了四个ＥＳ细胞系,分别命名为ＣＥ１、ＣＥ２、ＣＥ３、ＣＥ４（Ｆｉｇ．１ａ＆ｂ）。这四个细胞系核型正常率均达到７０％以上。我们检查了ＣＥ２细胞的分化能力,当将ＣＥ２细胞注入同基因型小鼠的皮下后,获得的畸胎瘤（Ｆｉｇ．３）组织切片检查的结果表明：该细胞能够分化成多种组织（Ｆｉｇ．２）。我们也研究了ＥＳ细胞的嵌合能力,用ＩＣＲ小鼠胚胎作为受体胚胎,采用囊胚显微注射法构建嵌合鼠。在幸存的幼鼠中我们获得了来源于ＣＥ２细胞的嵌合鼠（Ｔａｂｌｅ１,Ｆｉｇ．４）。综