Cytoplasmic genetic effects and growth of hybrid mice

Variation in cellular biochemical functions controlled by cytoplasmic genes was studied in relation to phenotypic differences between progeny of reciprocal hybrid female mice. Least squares procedures were used to test for differences in mitochondrial respiratory metabolism and in capacity for ATP synthesis, and differences in growth of progeny of hybrid dams. Under identical nuclear influences, mitochondria of A/J and C57BL/6J cytoplasms differed (P less than .10 to P less than .01) from those of BALB/cJ cytoplasm in energy conservation. No differences were detected in mitochondrial efficiency between BALB/cJ cytoplasm evaluated in different nuclear environments. Three-way cross progeny of C57BL/6J x BALB/cJ reciprocal hybrid females mated to DBA/2J males differed (P less than .05) in litter weight at weaning and 1 wk and 2 wk postweaning. The F2 progeny of reciprocal C57BL/6J x BALB/cJ dams and F2 and three-way cross progeny of reciprocal A/J x BALB/cJ dams did not differ in weight at any age measured. Across all genotypes of dam, rank correlations of mitochondrial traits with F2 litter weights were nonsignificant. Observed variation in mitochondrial functions partially controlled by cytoplasmic genes did not limit mouse growth under these experimental conditions.     

SPF级BALB/cC57BL/6小鼠繁殖性能及生长发育的比较研究

为提高SPF级实验动物的质量,选择10周龄BALB/c C57BL/6小鼠各50只（雌雄各半）,采取雌雄1？1近亲交配繁殖,并统计其生长发育和繁殖性能指标。BALB/c小鼠1～3周龄的平均窝重及离乳后的体重增长明显大于C57BL/6小鼠（p〈0.01）。第1胎交配分娩间隔,第1和第3胎离乳育成率品系间差异均有显著性（P〈0.05及P〈0.01）。BALB/c小鼠的体格比C57BL/6小鼠大,带乳能力比C57BL/6小鼠强但产仔能力比C57BL/6小鼠弱。     

Effects of diesel exhaust particles on the male reproductive system in strains of mice with different aryl hydrocarbon receptor responsiveness

Diesel exhaust particles (DEPs) contain polycyclic aromatic hydrocarbons (PAH) that bind to aryl hydrocarbon receptors (AhRs) and decrease sperm production. Since it is not clear if AhR mediates DEP toxicity, we investigated the effect of DEPs in four strains of mice that have different AhR responsiveness. We treated BALB/c, C57BL/6, ICR and DBA/2 mice with DEP suspensions and compared their toxicity in each strain. In both the vehicle- and DEP-treated groups, ethoxyresorufin-O-deethylase (EROD) activity, as an indirect index of AhR activity, was increased in the order of BALB/c > C57BL/6 > ICR > DBA/2. Only BALB/c and C57BL/6 mice had significantly lower daily sperm production (DSP) than vehicle-treated mice. All strains exhibited increased sperm abnormalities. In particular, the C57BL/6, ICR and DBA/2 mice exhibited significantly increased abnormalities. A significant correlation was found between EROD activity and DSP or incidence of morphologically abnormal sperm. These data suggest that DEP toxicity may affect the male reproductive system in an AhR-dependent manner.     

Encephalomyocarditis (EMC) virus-induced myocarditis by different virus variants and mouse strains.

The mode of occurrence of encephalomyocarditis (EMC) virus-induced myocarditis in mice was pathologically and virologically investigated using 2 virus variants (highly diabetogenic EMC-D and non-diabetogenic EMC-B) and 2 mouse strains (diabetes-susceptible BALB/c and diabetes-resistant C57BL/6). Mice were inoculated with 10(5) PFU/head of the virus intraperitoneally and observed up to 7 days post inoculation (7DPI). As compared with EMC-B-infected BALB/c and EMC-D-infected C57BL/6 mice, EMC-D-infected BALB/c mice developed marked myocarditis and exhibited a heart virus titer of more than 100 times above that of the others after 4DPI. Electron microscopically, small aggregations of virus-like particles, with 20-25 nm in diameter, were found in the cytoplasm of degenerated cardiomyocytes showing mitochondrial and myofibrillar degeneration in EMC-D-infected BALB/c mice.     

Fundamentals of host immune response against Brucella abortus: what the mouse model has revealed about control of infection

The studies reviewed here evaluated the role cellular immune system components play in control of brucellosis by conducting comparative studies with brucella-resistant C57BL/10 or C57BL/6 mice and susceptible BALB/c mice. We have shown by both in vitro and in vivo studies that activation of macrophages with interferon-gamma (IFN-γ) is an important factor for control of infection with B. abortus in the mouse model and that the mechanism of anti-brucella activity largely involved reactive oxygen intermediates. Differences in control of the organism by resistant and susceptible mice was not related to inherent differences in the ability of their macrophages to control infection either with or without IFN-γ activation nor was it attributable to NK cells since we found no role for them in control of brucellosis in either mouse strain. However, relative resistance to brucellosis did correlate with increased production of IFN-γ by CD4 T cells during the first weeks after infection while IL-10 contributed to susceptibility in BALB/c mice. Moreover, by 3 weeks post-infection splenocytes from the susceptible BALB/c mice failed to produce IFN-γ and relied on TNF- as well as CD8 T cells to control infection until the end of the plateau phase around 6 weeks post-infection when IFN-γ production resumed and clearance began. In contrast, IFN-γ was crucial for control throughout the infection in the more resistant C57BL/6 mice and the mice died in its absence by 6 weeks post-infection compared to 12 weeks for the more susceptible mice that relied on additional mechanisms of control. In contrast to the IFN-γ knock-out mice, both β2 microglobulin knock-out C57BL/6 mice, which do not express conventional MHC class I molecules and thus cannot present antigen to CD8 T cells, or perforin knock-out C57BL/6 mice, which have no T cell cytotoxic activity, controlled and cleared the infection as well as normal C57BL/6 mice. The hiatus of IFN-γ production in BALB/c mice correlated with very high levels of total IL-12 and it was postulated that the lack of IFN-γ was a consequence of p40 homodimer blocking activity. However, reduction of p40 IL-12 in vivo through administration of indomethacin reduced the infection without a concomitant measurable increase in IFN-γ. Current studies are aimed at elucidating the mechanism of the IFN-γ hiatus.     

Establishment of ES cells from inbred strain mice by dual inhibition (2i)

A number of mouse ES cells from inbred strains have been established to date, but efficiency varies across the different strains. The 129 strain mouse is efficient to establish, whereas C57BL/6 and BALB/c strains are not. It is possible that their genetic backgrounds account for the difference in their ability to establish ES cell lines. In this study, we attempted to establish C57BL/6J and BALB/c Cr ES cells by dual inhibition (2i) using two inhibitors (PD0325901 and CHIR99021) of extracellular signal regulated-kinase (ERK) and glycogen synthase kinase-3 (GSK-3), which promote ES cell differentiation. The results revealed that the establishment efficiencies of C57BL/6J and BALB/c Cr ES cells were remarkably increased by 2i. These ES cells stably expressed pluripotent markers and generated high-contribution chimeras with germline transmission. Furthermore, we generated germline chimeras from C57BL/6J ES cells through the method of gene modification. These findings indicate that 2i is a powerful tool for establishing C57BL/6J and BALB/c Cr ES cells with the ability to generate germline chimeras.     

Heme oxygenase-1 activity is involved in the control of Toxoplasma gondii infection in the lung of BALB/c and C57BL/6 and in the small intestine of C57BL/6 mice

Heme oxygenase-1 (HO-1) is an enzyme that catabolizes free heme, which induces an intense inflammatory response. The expression of HO-1 is induced by different stimuli, triggering an anti-inflammatory response during biological stress. It was previously verified that HO-1 is able to induce indoleamine 2,3-dioxygenase (IDO), an enzyme that is induced by IFN-γ in Toxoplasma gondii infection. To verify the role of HO-1 during in vivo T. gondii infection, BALB/c and C57BL/6 mice were infected with the ME49 strain and treated with zinc protoporphyrin IX (ZnPPIX) or hemin, which inhibit or induce HO-1 activity, respectively. The results show that T. gondii infection induced high levels of HO-1 expression in the lung of BALB/c and C57BL6 mice. The animals treated with ZnPPIX presented higher parasitism in the lungs of both lineages of mice, whereas hemin treatment decreased the parasite replication in this organ and in the small intestine of infected C57BL/6 mice. Furthermore, C57BL/6 mice infected with T. gondii and treated with hemin showed higher levels of IDO expression in the lungs and small intestine than uninfected mice. In conclusion, our data suggest that HO-1 activity is involved in the control of T. gondii in the lungs of both mouse lineages, whereas the hemin, a HO-1 inducer, seems to be involved in the control of parasitism in the small intestine of C57BL/6 mice.     

Pup mortality in laboratory mice – infanticide or not?

Background

Despite being the most commonly used mammal in biomedical research, problems with perinatal mortality in mice have received little attention and the causes of pup death are still poorly known. Females are often housed alone with their litters and since the lost pups are generally eaten, it is commonly assumed that the mother has killed them. However, more detailed observations than have been reported previously in the literature are required to establish if the cause of death is infanticide. Litter loss can only be prevented efficiently after underlying causes have been carefully investigated and interpreted. The aim of this study was to investigate if females actively kill their pups by observing the behaviour of females and pups in litters that later were lost. We used video recordings of females that lost their entire litter to observe females in detail from parturition until the pups died. In total, 10 C57BL/6 females (wildtype and the knockouts Hfe^−/− and β2m^−/−) were studied, housed in Makrolon II cages with or without access to a small amount of nesting material.

Results

Three of the females had pups that were never seen moving, and another three females had one or two pups that never moved, indicating that some pups were most likely still-born. In five females with live-born pups, detailed observations from the time when a pup was last seen moving until it died were possible to carry out. We observed females eating dead offspring and interacting with both moving and dead pups. However, we never observed a pup stop moving when manipulated by the female, nor were any wounds seen in the pups. Hence, we found no evidence of infanticide when studying females that had lost their entire litter.

Conclusion

These results suggest that other causes than infanticide plays a major role in mouse pup death, and stress the need for more systematic and careful investigations of the causality of litter loss.     

The local immune response of mice after Helicobacter suis infection: strain differences and distinction with Helicobacter pylori

Helicobacter (H.) suis colonizes the stomach of pigs and is the most prevalent gastric non-H. pylori Helicobacter species in humans. Limited information is available on host immune responses after infection with this agent and it is unknown if variation in virulence exists between different H. suis strains. Therefore, BALB/c and C57BL/6 mice were used to compare colonization ability and gene expression of various inflammatory cytokines, as determined by real-time PCR, after experimental infection with 9 different H. suis strains. All strains were able to persist in the stomach of mice, but the number of colonizing bacteria at 59 days post inoculation was higher in stomachs of C57BL/6 mice compared to BALB/c mice. All H. suis strains caused an upregulation of interleukin (IL)-17, which was more pronounced in BALB/c mice. This upregulation was inversely correlated with the number of colonizing bacteria. Most strains also caused an upregulation of regulatory IL-10, positively correlating with colonization in BALB/c mice. Only in C57BL/6 mice, upregulation of IL-1β was observed. Increased levels of IFN-γ mRNA were never detected, whereas most H. suis strains caused an upregulation of the Th2 signature cytokine IL-4, mainly in BALB/c mice. In conclusion, the genetic background of the murine strain has a clear impact on the colonization ability of different H. suis strains and the immune response they evoke. A predominant Th17 response was observed, accompanied by a mild Th2 response, which is different from the Th17/Th1 response evoked by H. pylori infection.     

Optimization of the use of C57BL/6 mice as a laboratory animal model for Neospora caninum vaccine studies

The development and testing of vaccines for Neospora caninum in mice require challenge studies to demonstrate a reduction in clinical signs or prevention of vertical transmission of the parasite after vaccination. Genetic susceptibility to N. caninum varies with the strain of mice. In this study, C57BL/6 mice were evaluated as a model for Neospora vaccine studies. A lethal challenge model was developed and the LD(50) was determined to be 1.5 x 10(7)N. caninum tachyzoites/mouse, delivered intraperitoneally. Brain lesions encountered in sections from sub-lethally challenged mice were scored on the basis of severity and total number of lesions to develop a histopathological scoring system for vaccine efficacy. A vertical transmission model for N. caninum vaccine studies was developed by studying mice that were infected either 2 weeks prior to mating or between days 12 and 14 of pregnancy. It was found that infection prior to mating reduced the average number of pups per litter. DNA extracted from fetal tissue was examined by a N. caninum specific polymerase chain reaction (PCR). The rate of vertical transmission was 0, 100 and 90.5% for the uninfected controls, mice infected during pregnancy and mice infected before mating, respectively. This study demonstrates that the C57BL/6 strain of mice is a good model for N. caninum vaccine studies because it is possible to establish a clear-cut lethal challenge model in C57BL/6 mice and they transmit the disease to their offspring efficiently.     

Different A-type natriuretic peptide level in five strains of mice

Atrial (A-type) natriuretic peptide (ANP) is vasodilative hormone involved in the regulation of blood pressure and volume homeostasis. In this study, we examined the differences of the auricular and plasma ANP distribution by immunohistochemistry, ultrastructural morphometry, and radioimmunoassay in five strains of mice. The ANP-immunoreactivities of the auricle were most intense in ICR, and moderate in C57BL/6J and C3H/HeN, and weakest in BALB/cA and DBA/2Cr. The number of ANP-granules was greatest in ICR followed by C57BL, C3H or BALB/c, and DBA/2 mice, in this order. The auricular ANP content was highest in ICR, but the plasma ANP concentration was comparable in all strains. The present study demonstrates that there are differences in the ANP circulating system between five strains.     

rasH2 mouse: reproducibility and stability of carcinogenicity due to a standardized production and monitoring system

The rasH2 mouse was developed as a model for carcinogenicity studies in regulatory science. Its phenotype is stable during high-volume production and over successive generations. To produce rasH2 mice, three strains of mice (C57BL/6J-TgrasH2, C57BL/6J, and BALB/cByJ) were maintained individually. Since the homozygous c-HRAS genotype is lethal, hemizygous transgenic mice were maintained by crossing with inbred C57BL/6J mice. After breeding, male B6-transgenic mice were mated with female BALB/cByJ mice to obtain transgenic mice. Pups that were rasH2-Tg (tg/wt) or rasH2-Wt (wt/wt) were confirmed by genotyping. Frozen embryos were preserved by the Central Institute for Experimental Animals (CIEA) and sent to two facilities, CLEA Japan and Taconic Biosciences, where the mice were produced. Production colonies are created in both facilities and supplied to customers worldwide. To prevent genetic drift, the colonies were renewed for up to 10 generations, and renewals were carried out four times every five years from 2005 to 2021. To ensure the uniformity and maintenance of the phenotype of rasH2 mice, the carcinogen susceptibilities were monitored in every renewal of colonies by CIEA based on a standard protocol of the short-term carcinogenicity study using the positive control compound N-methyl-N-nitrosourea (MNU). Furthermore, simple carcinogenicity monitoring targeting the forestomach, the organ most sensitive to MNU, was performed approximately once a year. Based on the optimally designed production and monitoring systems, the quality of rasH2 mice with reproducibility and stability of carcinogenicity is maintained and supplied globally.     

A fatty acid binding protein from Fasciola hepatica induced protection in C57/BL mice from challenge infection with Schistosoma bovis.

Three strains of mice (NMRI, C57/BL, BALB/c) were each immunized with a 12 kDa purified, native Fasciola hepatica fatty acid binding protein (Fh12) and challenged percutaneously with Schistosoma bovis cercariae. C57/BL mice immunized with Fh12 had significant reductions in S. bovis worm burden recoveries (96 and 87% reductions over controls in two separate experiments). When using NMRI or BALB/c mice, Fh12 alone or in Freund's adjuvant failed to induce significant protection against S. bovis. In C57/BL mice vaccinated against Fh 12, antibodies to the IgG2a isotype, but not to the IgG1 isotype, increased by 2 weeks after the second immunization and remained high through 8 weeks of S. bovis infection. Antibodies to S. bovis increased after 4 weeks of infection. Regarding cytokine production by spleen mononuclear cells, C57/BL mice vaccinated with Fh12 in adjuvant, and having the highest protective response against challenge infection with S. bovis, had an increase of IFN-gamma production with Concanavalin A but no increase of IL-4 in similarly stimulated cells. These results suggest that the protection obtained in this group of mice is mediated by a Th1 immune response.     

近交系小鼠RAPD标记的观察

采用随机扩增多态性DNA技术(RAPD),分析BALB/c、C57BL/6、DBA/2、C3H/He等4个近交系小鼠的基因多态性,探讨用RAPD作为遗传标记,对近交系小鼠进行遗传检测。结果表明,4种小鼠表现出了各自不同的多态性RAPD标记,证明RAPD可作为近交系小鼠的分子标记,在DNA水平区别4种近交系小鼠。     

A2a基因敲除小鼠生物学特性的初步研究

为了解A2a基因敲除小鼠的生物学物性,给研究心血管疾病及神经疾病提供部分基础资料,试验采用全自动生化分析仪及血细胞分析仪分别对60日龄C57BL/6、A2a雌雄小鼠的常用血液学指标及生化指标值进行检测,并测定了各小鼠的脏器。结果:C57BL/6和A2a小鼠血液的血小板计数、红细胞分布宽度差异显著(P0.05)。C57BL/6和A2a小鼠的丙氨酸转氨酶、肌酸激酶差异显著(P0.05),乳酸脱氢酶差异极显著(P0.01),脏器系数无显著差异(P0.05)。     

Infections in immunocompetent and immune-deficient mice with promastigotes of a North American isolate of Leishmania infantum

Leishmania infantum, an etiologic agent of zoonotic visceral leishmaniasis, is widespread among foxhounds in the United States. Experimental infections with a North American isolate of L. infantum were evaluated using two inoculation routes in immunocompetent and immunosuppressed mouse strains. Groups of 2-5 interferon gamma gene knockout (IFN-gamma-KO) (BALB/c-Ifng), inducible nitric oxide synthase (NOS) gene knockout (iNOS-KO) (C57BL/6), B-cell-deficient (microMT) (C57BL/6), and BALB/c mice were intravenously (i.v.) or subcutaneously (s.c.) inoculated with various doses of promastigotes of the LIVT-1 strain of L. infantum. None of the mice developed clinical signs of leishmaniasis during the 8-9 weeks of the study. Promastigotes were cultured from spleens of all i.v.-infected mice by 3 days post culture. Spleens from s.c.-infected mice inoculated with greater than 1 x 10(6) parasites became culture positive 3-24 days post culture, but promastigotes were not cultured from mice infected with 1 x 10(5) or 5 x 10(5) LIVT-1 promastigotes. Histological lesions were prominent in the livers of i.v.-infected mice but were mild to nonexistent in s.c. infection. Serological responses were low and transient determined by indirect fluorescent antibody testing in all groups. These results indicate that the i.v. route of infection is superior to the s.c. route in a mouse model of North American leishmaniasis and that mice lacking INF-gamma, iNOS or mice that are B-cell-deficient are not more susceptible to acute infection.     

Interleukin-6 is produced during both murine and avian Eimeria infections

The production of interleukin-6 (IL-6) during Eimeria infection was investigated in an attempt to gain a better understanding of the role of this multi-functional cytokine in resistance to this parasite. IL-6 production was measured in both chickens, in which the disease is of economic importance, and the better-characterised murine model system.Systemic and local IL-6 production in mice during E. vermiformis infection was investigated, in the relatively resistant BALB/c strain, and the relatively susceptible C57 BL/6 strain, using a murine IL-6 ELISA and the 7TD1 assay. Enhanced systemic production of IL-6 in serum was seen in infected BALB/c mice when compared to C57 BL/6 mice. This difference was also reflected in the draining lymph node of the site of infection, assessed by testing supernatants from stimulated mesenteric lymph node cells taken from infected mice at different times post-infection.Production of chicken IL-6-like factor activity was investigated using a murine IL-6 7TD1 bioassay. The presence of substantial quantities of IL-6-like factor activity was detected in serum taken from some chickens infected with E. tenella during the course of primary infection and, in a separate experiment, during the first few hours post-infection, a time when the pro-inflammatory capacity of IL-6 would influence the developing immune response. These results suggest that IL-6 is also important in the induction of immune effector responses to Eimeria infections in the chicken.     

Neonatal-piglet weight variation and its relation to pre-weaning mortality and weight gain on commercial farms

To determine the effect of within-litter neonatal-weight variation on pre-weaning mortality and weight gain, we analyzed piglet survival and weight gain within 400 litters from 10 commercial farms. Neonatal-weight variation (independent of mean neonatal weight, litter size and sow parity) was associated with pre-weaning survival and weaning-weight variation-but not with mean weaning weight. Neonatal piglets with weights well below the range of most of the litter (low-birth-weight piglets) had an increased risk of dying and were unable to obtain normal weight gains by weaning if they survived. These piglets experienced lower survival and poorer weight gain in larger litters. These piglets also tended to have lower survival but normal (albeit low) weaning weights if they survived in litters from middle-aged and old sows. High neonatal-weight variation resulted in lower survival and more variable weaning weights. Small piglets had a greater risk for poor survival and weight gain compared to their heavier litter-mates (a disadvantage that was exacerbated in large litters).     

Effect of birth and fraternal litter size and cross-fostering on growth and reproduction in swine

Twelve hundred fifty-one pigs from six farrowings (FGRP) were classified within a FGRP by their birth litter size (BL- = below average and BL+ = above average), randomly allotted to nursing litter sizes of 6 or 12+ pigs/sow (NL- vs NL+) and reared by their own or foster dams (XF- vs XF+). Pigs were weighed at birth, 21 d and when near 105 kg. A random sample of 40 gilts per FGRP was retained for observation of pubertal age and primipara conception. Twenty-four gilts per FGRP were farrowed and rebred for a second parity. Pigs born in large litters were younger at 105 kg than those born in small litters (189 vs 196 d +/- 1.4); no other differences (P greater than .05) were observed for BL. Pigs reared in larger litters had lower survival rate from birth to weaning (79 vs 86% +/- 1), had slower weight gains to 21 d of age (5.3 vs 6.6 kg +/- .17) and were older at 105 kg (195 vs 190 d +/- 1.4) than those reared in small litters (P less than .04). Cross-fostered pigs were slower gaining to 21 d (5.9 vs 6.1 kg +/- .14) and were older at 105 kg (195 vs 191 d +/- 1.4) than pigs not cross-fostered pigs (P less than .02). Growth beyond 105 kg and pubertal age were unaffected by any factor studied (P greater than .05). Although size of birth litter did not affect (P greater than .05) any reproductive trait, an interaction between litter size and farrowing group was detected.(ABSTRACT TRUNCATED AT 250 WORDS)     

胎次与配种月份对大约克母猪繁殖性能的影响

对河南省某原种猪场2002～2004年270窝法系大约克母猪各胎产仔数进行统计,根据不同胎次、不同配种月份分别研究法系大约克母猪的繁殖性能。结果表明：胎次对产仔数的影响较大,其产仔数性能以第一胎低,随胎次增加,繁殖性能逯步上升,至第五胎达到高峰,此后逯渐下降;配种月份对其繁殖性能的影响不大,但以10月份配种的母猪繁殖性能最差。