Cytofluorimetric study of ploidy in the preleukemic thymus in C57BL/Ka mice under fractionated irradiation

The ploidy of the thymus was studied in C57B1/Ka mice irradiated with 4 weekly X-Ray doses of 1.75 Gy. The determination of nuclear DNA content was performed by flow cytometry of intact thymocytes labeled with propidium iodide in presence of a mixture of chicken and rainbow trout red blood cells as internal reference standards. The method has been tested by detecting the sex difference in DNA content of G0/G1 of normal thymic mouse cells. The mean value was 2.9% higher in female mice. The thymus of almost 60% of irradiated male mice present a slight hypoploidy of 2.6% one month after the last irradiation.     

Interaction of radiation-induced preleukemia cells and bone marrow grafts in C57BL/Ka mice

The repopulation of thymus inoculated with radiation-induced preleukemia cells was studied in 400 R irradiated mice grafted with normal bone marrow cells. These marrow cells gave rise to an actively regenerating thymic progeny, as well as in 400 R treated mice receiving only a bone marrow graft. Moreover, the marrow graft did not prevent the progression of inoculated preleukemic cells towards lymphoma growth.     

Migration of bone marrow cells to the thymus in mice after a single sublethal-dose irradiation

In sublethally irradiated mice, thymus repopulation is due first to the proliferation of surviving thymocytes followed by the multiplication of bone marrow derived prothymocytes. The migration of bone marrow cells to the thymus after a single sublethal whole-body X irradiation was studied by using fluorescein isothiocyanate as a cell marker. Irradiation increases the permissiveness of the thymus to the immigration of bone marrow cells. Furthermore, the post-Rx regenerating bone marrow cells exhibit migration capacities greater than the normal ones. The radiation induced changes in the bone marrow thymus interaction might play an important role in thymus regeneration after sublethal irradiation.     

NK activity in C57BL/Ka mice during the development of radiation-induced lymphomas

Treatment of C57BL/Ka mice with a split dose wholebody irradiation (four weekly irradiations of 1,75 Gy) induces the development of thymic lymphomas. NK activity of spleen cells has been determined at several intervals after leukemogenic treatment. Two days after irradiations. NK activity is normal and decreases strongly after one week. This period of decline persists during about one month. Then, NK activity restores and reaches control values. Lymphomas appear in spite of NK activity restoration. The diminution of NK activity during the preleukemic period could favour preleukemic cells apparition.     

Thiophosphamide-induced chromosome aberrations in embryonal liver and bone marrow cells of A/He and C57BL/L mice]

Chromosomal aberrations were studied in cells of embryonic liver and bone marrow of A/He and C57BL/6 mice following mutagenic treatment by the alkylating agent thiophosphamide in g1-S periods of the cell cycle. Higher sensitivity of chromosomes to aberration induction was found in A/He mice.     

Tumours of the bone in C57BL/10J mice

Tumours of the bone were found in 17 (12 females and 5 males) of 6768 (0.25%) C57BL/10J mice (3384 males and 3384 females) pooled from 14 oncogenic studies carried out from 1973 to 1987. Tumours of bone were found predominantly in female mice and were age related. The tumours were classified histologically as osteosarcoma (5 osteoblastic, 6 mixed type and 1 fibroblastic), chondroma (2), chondromyxoid fibroma (1), chondrosarcoma (1) and periosteal fibroma (1).     

Embryonic stem cells derived from C57BL/6J and C57BL/6N mice

Mouse embryonic stem (ES) cells with the C57BL/6 genetic background allow the generation of knockout mice without the need to backcross to C57BL/6. However, C57BL/6 ES cells whose pluripotency after homologous recombination has been confirmed are not yet available from public cell banks. To facilitate the use of ES cells derived from C57BL/6 sublines in both biologic and medical research, we demonstrated that the use of knockout serum replacement as a medium supplement and 8-cell blastomeres as recipient embryos allowed establishment of ES cells and production of germline chimeric mice, respectively. Under effective conditions, a large number of ES cell lines were established from C57BL/6J and C57BL/6N blastocysts. The majority of ES cells in many cell lines obtained from both strains showed a normal chromosome number. Germline chimeric mice were generated from C57BL/6J and C57BL/6N ES cells. Finally, the ES cell line B6J-S1UTR, derived from C57BL/6J, was used for successful production of gene knockout mice. C57BL/6J ES (B6J-S1UTR and B6J-23UTR) and C57BL/6N ES (B6N-22UTR) cells are available from the cell bank of the BioResource Center at RIKEN Tsukuba Institute (http://www.brc.riken.jp/lab/cell/english/).     

Protective effect of bone marrow and spleen suspensions on radiation-induced leukemogenesis in C57BL/6 mice

Summary The present experiments are an attempt to precise the type and localization of the cells involved in the protective effect of hemopoietic suspensions against the radiation-induced thymic lymphosarcoma (TLS) of C57BL/6 mice. Inocula containing variable numbers of BM or spleen CFUs from 60-day-old and 360-day-old donors were tested. According to their origin, the suspensions differed with respect to the CFU replication rate, the CFU ability to differentiate towards the T lineage and the content of the suspensions in thymic precursors. Two levels of inhibition were observed: BM suspensions from 60-day-old donors containing 1,500 CFUs had the best protective effect: 14.5% of TLS; 1,500 CFUs from 360-day-old donors were slightly but not significantly less efficient (28.5%). The second level of inhibition (36–46% of TLS) was obtained with all the following inocula:a) 1,200 and 300 spleen CFUs or 300 and 95 BM CFUs from 60-day-old donors,b) 1,500 spleen CFUs from aged donors. Seventy-six spleen CFUs from 60-day-old donors, 120 BM or 175 spleen CFUs from aged donors had no effect. These results suggest that in addition to the high replication rate of the BM CFUs as compared with spleen CFUs, cells endowed with an optimal protective effect are present in BM suspensions and are either absent or present in very small amount in spleen suspensions. These cells which induce an early repopulation of the thymus might correspond to thymic precursors.     

Relations between the capacity of thymic repeopling and the formation of lymphoma obtained by radiation-induced preleukemic cells in C57BL/Ka mice

We have analyzed the repopulation of thymuses injected with preleukemic cells obtained at various interval after fractionated irradiation. Our results show that preleukemic cells can repopulate the thymus transiently as normal thymocytes or they can proliferate in recipients. In all cases, preleukemic cells can give rise to donor lymphomas.     

Mast cells and macrophages in normal C57/BL/6 mice

Mast cells and macrophages have an important role in immunity and inflammation. Because mice are used extensively for experimental studies investigating immunological and inflammatory responses, we examined mast cell and macrophage distribution in normal murine tissues. Mast cells were abundant in the murine dermis, tongue, and skeletal muscle but were rarely found in the heart, lung, spleen, kidney, liver, and the bowel mucosa. In contrast, dogs exhibited large numbers of mast cells in the lung parenchyma, liver, and bowel. Some murine dermal mast cells had long cytoplasmic projections filled with granular content. Mouse mast cells demonstrated intense histamine immunoreactivity and were identified with histochemical enzymatic techniques for tryptase and chymase. Macrophages, identified using the monoclonal antibody F4/80, were abundant in the spleen, lung, liver, kidney, and bowel but relatively rare in the heart, tongue, and dermis. Using a nuclease protection assay we investigated mRNA expression of stem cell factor (SCF), a crucial survival factor for mast cells, and the macrophage growth factors macrophage colony stimulating factor (M-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF). Stem cell factor mRNA was highly expressed in the murine lung. Relatively low levels of SCF mRNA expression were found in the tongue and earlobe, which are tissues containing a high number of mast cells. Macrophage CSF and GM-CSF mRNA was highly expressed in the lung and spleen. The murine heart, an organ with a low macrophage content, expressed high levels of M-CSF but negligible levels of GM-CSF mRNA. Constitutive growth factor mRNA expression in murine tissues without significant populations of mast cells and macrophages may suggest an alternative role for these factors in tissue homeostasis.     

Lympho-epithelial interactions in the thymus during development of radio-induced lymphomas in C57BL mice

In C57BL/Ka mice, leukemogenic fractionated whole-body X-irradiation induces alterations of the lymphoepithelial interactions normally found in the Thymic Nurse Cells (TNCs) and leads to the disappearance of these complexes. This phenomenon is due to the disturbances of thymic lymphopoiesis caused by modifications of bone marrow prothymocytes and of the epithelial component of TNCs.     

Characteristics of tumors developed in mdx mice after transplantation of GFP-positive mesenchymal stem cells isolated from bone marrow of transgenic C57BL/6 mice

The purpose of the work was morphological and histochemical examination of tissue differentiation in tumors developed in mdx mice after the intramuscular transplantation of GFP-positive mesenchymal bone-marrow stem cells (MSC-GFP) derived from C576BL/6 transgenic mice and cultivated for 43–45 passages. These cells did not generate tumors in syngeneic adult C57BL/6 mice. The tumors were classified as mesenhymomas, fibrosarcomas, and sarcoma. Adipocyte and chondrocyte clusters, as well as bone areas with erythroid, myeloid, and thrombocyte hematopoiesis and neural tissue with glia cells were observed inside of tumors. Types of tissue tumor differentiation were similar to those described in the literature for MSCs induced to differente in vitro by specific treatment. However, the differentiation spectrum in MSC-GFP-produced tumors was broader than the differentiation of tumors derived from adult mouse MSCs spontaneously transformed or transfected in vitro. The results presented here, along with our previous data, demonstrate that the transfection of stem cells, including totipotent stem cells, with genetic constructs is accompanied by the destabilization of the cell genome, even if the activity of inserted gene (GFP) does not affect general cell functioning.     

Effect of autologous marrow transplantation on bone marrow recovery following large-field fractionated irradiation of dogs

After large-field fractionated irradiation of dogs at a cumulative dose of 54 Gy, a stable bone marrow depletion occurs persisting for a year following irradiation. The automyelotransplantation after the end of the exposure elicits a transient recovery of the exposed bone marrow, 1.5-2 months after the beginning of irradiation, followed by a secondary depletion of the exposed haemopoietic sites. The control and the automyelotransplanted animals exhibited bone marrow recovery one year and six months after irradiation, respectively, the cellularity being maintained at a high level for 3 years of observation.     

Differences in anxiety-related behavior between apolipoprotein E-deficient C57BL/6 and wild type C57BL/6 mice

The influence of ApoE gene deletion on the anxiety state has not been previously investigated. The elevated plus maze was used in this study to determine differences in anxiety-related behavior between apoE-deficient and wild type C57BL/6 mice. The apoE-deficient mice demonstrated less anxiety on the elevated plus maze by spending more time in the open arms of the elevated plus maze compared to wild type mice (p<0.001). Additionally, female apoE-deficient mice visited the open arm of the maze more often than their apoE-deficient male counterpart (p<0.05). The anxiety state and/or sex are possible variables to be considered when designing physiological and/or behavioral studies involving mice that are apoE-deficient.     

Enhanced susceptibility to lupus contributed from the nonautoimmune C57BL/10, but not C57BL/6, genome

Genes from New Zealand Black and New Zealand White mice have been implicated in the development of a disease similar to human systemic lupus erythematosus. In an attempt to define the MHC class II genes involved in disease, we previously studied similarly designed backcrosses of New Zealand Black mice with C57BL/6 (B6) mice transgenic for Ez genes or with C57BL/10 (B10) mice transgenic for Az genes. Although the transgenes showed no effect on the development of autoantibody production or lupus nephritis in either backcross, surprisingly, there was greatly increased expression of these disease traits in the backcrosses involving B10 compared with B6 mice. These studies therefore implicated genetic contributions in B10 vs B6 backgrounds, despite their 98% identity. A genome-wide linkage analysis uncovered a B10 locus on mid-chromosome 13, which enhanced nephritis and was strongly linked with the production of pathogenic retroviral gp70-anti-gp70 immune complexes when contributed by B10, but not B6, mice. The subsequent identification of a single marker polymorphic between B10 and B6, along with the extreme genetic similarity between the two strains in this region, is likely to permit expedited identification of the lupus-susceptibility gene from this nonautoimmune strain.     

A comparative phenotypic and genomic analysis of C57BL/6J and C57BL/6N mouse strains

Background

The mouse inbred line C57BL/6J is widely used in mouse genetics and its genome has been incorporated into many genetic reference populations. More recently large initiatives such as the International Knockout Mouse Consortium (IKMC) are using the C57BL/6N mouse strain to generate null alleles for all mouse genes. Hence both strains are now widely used in mouse genetics studies. Here we perform a comprehensive genomic and phenotypic analysis of the two strains to identify differences that may influence their underlying genetic mechanisms.

Results

We undertake genome sequence comparisons of C57BL/6J and C57BL/6N to identify SNPs, indels and structural variants, with a focus on identifying all coding variants. We annotate 34 SNPs and 2 indels that distinguish C57BL/6J and C57BL/6N coding sequences, as well as 15 structural variants that overlap a gene. In parallel we assess the comparative phenotypes of the two inbred lines utilizing the EMPReSSslim phenotyping pipeline, a broad based assessment encompassing diverse biological systems. We perform additional secondary phenotyping assessments to explore other phenotype domains and to elaborate phenotype differences identified in the primary assessment. We uncover significant phenotypic differences between the two lines, replicated across multiple centers, in a number of physiological, biochemical and behavioral systems.

Conclusions

Comparison of C57BL/6J and C57BL/6N demonstrates a range of phenotypic differences that have the potential to impact upon penetrance and expressivity of mutational effects in these strains. Moreover, the sequence variants we identify provide a set of candidate genes for the phenotypic differences observed between the two strains.