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At least 829 records · Page 46Linked to original sources
PubMed1984

Mouse trisomy 16 as an animal model of human trisomy 21 (Down syndrome): production of viable trisomy 16 diploid mouse chimeras.

We have previously proposed that mice trisomic for chromosome 16 will provide an animal model of human trisomy 21 (Down syndrome). However, the value of this model is limited to some extent because trisomy 16 mouse fetuses do not survive as live-born animals. Therefore, in an effort to produce viable mice with cells trisomic for chromosome 16, we have used an aggregation technique to generate trisomy 16 diploid (Ts 16 2n) chimeras. A total of 79 chimeric mice were produced, 11 of which were Ts 16 2n chimeras. Seven of these Ts 16 2n mice were analyzed as fetuses, just prior to birth, and 4 were analyzed as live-born animals. Unlike nonchimeric Ts 16 mouse fetuses which die shortly before birth with edema, congenital heart disease, and thymic and splenic hypoplasia, all but 1 of the Ts 16 2n animals were viable and phenotypically normal. The oldest of the live-born Ts 16 2n chimeras was 12 months old at the time of necropsy. Ts 16 cells, identified by coat color, enzyme marker, and/or karyotype analyses, comprised 50-60% of the brain, heart, lung, liver, and kidney in the 7 Ts 16 2n chimeric fetuses and 30-40% of these organs in the 4 live-born Ts 16 2n animals. Ts 16 cells comprised an average of 40% of the thymus and 80% of the spleen in the Ts 16 2n chimeras analyzed as fetuses, with no evidence of thymic or splenic hypoplasia. However, we observed a marked deficiency to Ts 16 cells in the blood, spleen, thymus, and bone marrow of live-born Ts 16 2n chimeras as compared to 2n 2n controls. These results demonstrate that although the Ts 16 2n chimeras were, with one exception, viable and phenotypically normal, each animal contained a significant proportion of trisomic cells in a variety of tissues, including the brain. Furthermore, our results suggest that although the abnormal development of Ts 16 thymus and spleen cells observed in Ts 16 fetuses is largely corrected in Ts 16 2n fetuses, Ts 16 erythroid and lymphoid cells have a severe proliferative disadvantage as compared to diploid cells in older live-born Ts 16 2n chimeras. Ts 16 2n chimeric mice will provide a valuable tool for studying the functional consequences of aneuploidy and may provide insight into the mechanisms by which trisomy 21 leads to developmental abnormalities in man.

Animals↗
PubMed1989

Somatic cell-oocyte interactions in mouse oogenesis: stage-specific regulation of mouse oocyte protein phosphorylation by granulosa cells.

The relative rate of synthesis of a number of proteins and the protein phosphorylation pattern of growing and fully grown oocytes were influenced by the presence of granulosa cells. In particular, a 74-kDa phosphorylated protein was detected only in granulosa cell-enclosed growing mouse oocytes. When reaggregated with granulosa cells, the growing oocyte displayed the phosphorylated form of the 74-kDa protein but when oocytes were cultured on Sertoli cell monolayers or in granulosa cell-conditioned medium the 74-kDa protein was not phosphorylated. We propose that (1) granulosa cells regulate protein phosphorylation in mouse oocytes; (2) a 74-kDa protein is phosphorylated only in growing oocytes when surrounded by granulosa cells; and (3) granulosa cells, but not Sertoli cells, are competent to send the appropriate "signal" to the growing oocyte.

Animals↗
PubMed1984

Supermelanotic hybrids between mouse teratocarcinoma and mouse melanoma cells.

Each of four kinds of teratocarcinoma cells, OTT6050P, PCC4, PSA1 and LT, derived from 129 or LT mouse strain, was fused with B16-CAPr melanoma cells derived from C57BL/6J by using Sendai virus. The resultant hybrids were morphologically melanotic melanoma cells which were larger and more heavily pigmented than the parental B16-CAPr melanoma cells. The chromosome analysis and GPI electrophoresis demonstrated that all hybrids were products of fusion between a single teratocarcinoma cell and a single melanoma cell. The pigmentation in the hybrids between a 129 teratocarcinoma cell and a melanoma cell was much stronger than that in hybrids between an LT teratocarcinoma cell and a melanoma cell. This phenomenon was consistent with the difference of coat color between 129 and LT mouse strain. From these results, it was suggested that the genes of teratocarcinoma cells involved in the pigmentation are activated in the hybrids with B16-CAPr melanoma cells.

Animals↗
PubMed1988

Cataract in senescence accelerated mouse (SAM). 2. Development of a new strain of mouse with late-appearing cataract.

A new strain of mouse with late-appearing hereditary cataract was developed in the Senescence Accelerated Mouse (SAM-R/3). At the beginning of the selection of the new strain (generation 4), 5% of mice which lived over 20 weeks had cataract, at least unilaterally. During selection and brother-sister inbreeding, the incidence increased and reached 68% of 78 mice whose lifespan exceeded 20 weeks, in the 12th generation. In recent populations, (generations 9-14), the cataract began to appear around 10 weeks of age and about 81% of 64 females and 49% of 47 males had cataract after 32 weeks of age. Inflammatory lesions of the cornea and eyelids began to occur later in life and the incidence was lower than that of cataract. This observation suggested to us that the cataract of this new strain was not congenital in origin but rather was age-related and did not occur consequently after the onset of inflammatory lesions around the lens tissue. Stereomicroscopic examination of the lens revealed that many of these cataracts occurred initially at the posterior area and that the mature cataract showed a characteristic protrusion at the posterior pole and nuclear dislocation. Histology of the mature cataract also revealed protrusion of the posterior pole and showed degeneration and liquefaction of lens cortex, nuclear dislocation and destruction of posterior lens capsule. The wet weight, water and protein contents showed that the lens of this strain developed normally and that the untoward events were the result but not the cause of cataract. The SAM-R/3 strain should prove to be a suitable murine model for investigation of age-related changes in the lens, and cataractogenesis.

Aging↗
PubMed1984

Characterization of a factor inducing differentiation of mouse myeloid leukemic cells purified from conditioned medium of mouse Ehrlich ascites tumor cells.

A factor inducing differentiation of mouse myeloid leukemic cells (MI) into macrophages was purified to apparent homogeneity from 168 1 of CM of Ehrlich ascites tumor cells. The purified factor was half-maximally active at 2 X 10(-11) M. The factor was analyzed by radioiodination, SDS-polyacrylamide gel electrophoresis and autoradiography. Its Mr was 40 000-50 000. On reduction, the factor lost activity, but showed no subunit structure. Treatment of the factor with endo-beta-N-acetylglucosaminidase F, but not endo-beta-N-acetylglucosaminidase H, gave rise to a molecule of Mr 20 000-28 000. The activity of the factor from Ehrlich cells was completely neutralized by antiserum to the factor of Mr 50 000-70 000 from mouse fibroblast L929 cells.

Animals↗
PubMed1992

Cloning and sequencing of mouse collagenase cDNA. Divergence of mouse and rat collagenases from the other mammalian collagenases.

Mouse collagenase cDNA was cloned and sequenced. The deduced amino acid sequence was compared to those of the other mammalian collagenases and related matrix metalloproteinases. These comparisons, as well as those of some enzymatic properties, show that the rodent (mouse and rat) interstitial collagenases are very similar but differ more from the other interstitial collagenases than does human neutrophil collagenase. This supports the hypothesis that the order Rodentia is an outgroup to the other eutherian (placental) mammalian orders.

Amino Acid Sequence↗
PubMed1975

A preparation of Fc fragment of normal mouse IgG1 for production of rabbit anti mouse IgG1 serum.

Ascitic fluid globulins obtained by peritoneal irritation from normal CBA mice were digested with papain, and the digests were examined by gradient chromatography on CM cellulose. The classical three peaks of OD280 found by Porter with papain digests of rabbit IgG were obtained. Fraction II contained material which reacted in immunodiffusion against mouse anti-IgG. The ascending part of this peak reacted against anti-IgG1 and not against anti-TgG2 reacting material appearing at detectable levels in the descending part of this peak. A pool from the ascending part of this peak, containing Fc of IgG1 and not visibly contaminated with Fc of IgG2, was injected into rabbits. The resulting antisera were free enough of anti-IgG2 to be effective in removing IgG1 from alloantibody-containing globulins without appreciable loss of antibodies of IgG2 class, thus allowing the complement-dependent IgG2-class antibodies full expression of their titer, without competition by Ig1 class antibodies. Chromatography of native mouse globulin did not produce a similar degree of separation of Ig1 from IgG2.

Animals↗
PubMed1983

Stimulation of [3H]thymidine uptake in mouse marrow by granulocyte-macrophage colony stimulating factor from mouse lung conditioned medium.

Stimulation of [3H]thymidine uptake in mouse marrow cells by a haematopoietic factor, granulocyte-macrophage colony stimulating factor from mouse lung conditioned medium, was used to follow the activity of the factor in the medium, and during its partial purification from the medium. The assay was performed in microtitre plates and found to be much easier and faster than conventional colony counting. The marrow cells were incubated in flat-bottomed plates in the presence of the factor for 5 days before labelling with [3H]thymidine (2 muCi/well, 5 Ci/mmole) for 6 h. Stimulation of the [3H]thymidine uptake was compared with the development of granulocyte-macrophage colonies in semisolid methylcellulose culture. The activity followed by both assays showed identical behaviour when subjected to ammonium sulphate precipitation, Sephadex gel filtration, concanavalin-A-Sepharose chromatography and polyacrylamide gel electrophoresis.

Animals↗
PubMed1994

Bovine colostrum ultrafiltrate: an effective supplement for the culture of mouse-mouse hybridoma cells.

An ultrafiltrate fraction (UF) of bovine colostrum has been successfully used as a cell culture supplement for growth and monoclonal IgG antibody production of cultured mouse-mouse hybridomas derived from spleen cells. In this study we compared the ability of UF to support growth and antibody production of IgA hybridomas derived from Peyer's patch cells with that of an IgG hybridoma cell line. One IgG (LPC2) and two IgA hybridoma cell lines (RB3 and P2E7) were used as models. The optimal UF concentration for Ig production and cell growth for both the IgA hybridoma RB3 and the IgG hybridomas was 5-10%. Initial plating density was found to be a critical factor for IgA hybridoma cell growth: the IgA hybridomas required a seeding density of at least 70,000 cells/ml to grow compared to 15,000 IgG hybridoma cells/ml (Pakkanen et al., 1992). The addition of small amounts (up to 2%) of FBS in 10% UF supplemented medium did not enhance IgA production or cell growth. RB3 and LPC2 cells seeded at equal density and grown in 10% UF for 8 days attained maximum cell densities at 3-4 days that were 58% (RB3) or 34% (LPC2) lower than those in 10% FBS, but the total amounts of monoclonal antibody produced were 73% and 83%, respectively, of that in 10% FBS. Thus, Ig production per cell was 22-27% higher in 10% UF than in 10% FBS. Hybridoma cells could be cultured for at least 5 weeks without any reduction in growth rates, if medium was partially but not completely replaced twice a week. This suggests that hybridoma cells maintained in UF supplemented medium secrete growth promoting factors. Cells maintained in UF for up to 5 weeks sustained similar monoclonal antibody production rates as in short term culture. These results show that UF can be used as an economical and effective hybridoma culture supplement for the production of both IgG and IgA antibodies.

Animals↗
PubMed1987

Studies on radiation-induced chromosome aberrations in mouse spermatocytes. II. Dose-response relationships of chromosome aberrations induced at zygotene stage in mouse primary spermatocytes following fast neutron- and 60Co gamma-irradiations.

The chromosome aberrations induced at zygotene stage in mouse spermatocytes following exposures to fast neutrons and 60Co gamma-rays were examined at diakinesis-metaphase I. The dose-response relationships were well fitted to linear equation for deletion-type aberrations and to linear-quadratic equation for exchange-type aberrations in 60Co gamma-irradiation group. In fast neutron-irradiation group, the dose-response relationships were well fitted to linear equations for deletion- and exchange-type aberrations. The rate of deletion-type aberrations was remarkably high for fast neutrons, about 6 times higher than that after 60Co gamma-irradiation. The main types of chromosome aberrations observed were iso-chromatid breaks or fragments and chromatid exchanges in both irradiation groups as well as X-irradiation. These results indicate that there is a possibility that two double-strand breaks are induced simultaneously at iso-locus position in sister chromatids by a single track of radiations. Production of such single-track-induced two double-strand breaks in iso-chromatids may be very frequently expressed as iso-chromatid-type deletions in the high LET fast neutron-irradiation group. On the contrary, in the low LET 60Co gamma- or X-irradiation group, the above-mentioned mechanism may not be so effective for contribution to chromosome aberration induction in mouse spermatocytes. This mechanism was discussed in detail.

Animals↗
PubMed1991

Recommended protocols based on a survey of current practice in genotoxicity testing laboratories: III. Cell transformation in C3H/10T1/2 mouse embryo cell, BALB/c 3T3 mouse fibroblast and Syrian hamster embryo cell cultures.

A standardized protocol and guidelines for the performance of cell transformation testing in mouse embryo (C3H/10T1/2), mouse fibroblast (BALB/c 3T3) and Syrian hamster embryo (SHE) cells have been developed. The protocol is based primarily on current laboratory practices as determined by responses to a detailed questionnaire completed by North American and European governmental, university and contract laboratories involved with cell transformation experimentation. This report identifies those modifications to previously described methodologies which are being used on a regular basis and also serves to clarify confusing or inconsistent practices.

Animals↗
PubMed1994

Isolation of micronuclei from mouse blood and fluorescence in situ hybridization with a mouse centromeric DNA probe.

Spontaneously existing and chemically induced micronuclei were isolated from mouse blood. 50 microliters of cardiac blood was diluted with PBS and centrifuged. After this, the cell pellet was subjected to hypotonic treatment, fixed with acetic acid-methanol (1:3), and the lysate was filtrated through a 2-microns polycarbonate nucleopore membrane. Isolated micronuclei were air-dried on a glass slide and subjected to fluorescence in situ hybridization (FISH) using a mouse centromeric gamma satellite probe. Approximately half of the micronuclei isolated from vehicle control mice showed centromere signal(s). In these preliminary studies, the proportion of centromere-positive micronuclei was increased by treatment with spindle poisons (colchicine and vinblastine sulfate), decreased only slightly by 1-beta-D-arabinofuranosylcytosine, and was generally unaffected by mitomycin C.

Animals↗
PubMed1987

Suppression of in vitro antibody production by dimethylnitrosamine in mixed cultures of mouse primary hepatocytes and mouse splenocytes.

The suppression of in vitro antibody responses by dimethylnitrosamine (DMN) was produced in a mouse hepatocyte and splenocyte co-culture system. Mouse hepatocytes were isolated from female B6C3F1 mice and cultured for 20-24 hr to allow the formation of a monolayer on collagen-coated plastic petri dishes. Spleen cells were isolated from the same hybrid and were co-cultured with the hepatocytes along with DMN. Cyclophosphamide (CP), an immunosuppressive agent requiring metabolic activation that was included as an initial positive control, produced a marked suppression of the in vitro antibody responses to LPS, DNP-Ficoll, and SRBCs in 4 hr in the co-culture system. Under comparable conditions DMN markedly suppressed the response to SRBCs, marginally suppressed the response to DNP-Ficoll, and did not suppress the polyclonal response to LPS. The suppression by DMN was related to the rocking speed during the 4-hr co-culture period and was optimally produced when the cultures were not rocked. Addition of serum into the medium (10% fetal calf serum) during the co-culture period did not change the effects of DMN on the antibody response. However, the addition of extracellular DNA (1 mg calf thymus DNA/ml) prevented the suppression of the antibody response by DMN. These results suggest that DNA represents the primary macromolecular target for the reactive intermediate of DMN, and indicate that a syngeneic co-culture system can be used to characterize the in vitro immunosuppression

Animals↗
PubMed1988

Genome sequences of a mouse-avirulent and a mouse-virulent strain of Ross River virus.

The nucleotide sequence of the genomic RNA of a mouse-avirulent strain of Ross River virus, RRV NB5092 (isolated in 1969), has been determined and the corresponding sequence for the prototype mouse-virulent strain, RRV T48 (isolated in 1959), has been completed. The RRV NB5092 genome is approximately 11,674 nucleotides in length, compared with 11,853 nucleotides for RRV T48. RRV NB5092 and RRV T48 have the same genome organization. For both viruses an untranslated region of 80 nucleotides at the 5' end of the genome is followed by a 7440-nucleotide open reading frame which is interrupted after 5586 nucleotides by a single opal termination codon. By homology with other alphaviruses, the 5586-nucleotide open reading frame encodes the nonstructural proteins nsP1, nsP2, and nsP3; a fourth nonstructural protein, nsP4, is produced by read-through of the opal codon. The RRV nonstructural proteins show strong homology with the corresponding proteins of Sindbis virus and Semliki Forest virus in terms of size, net charge, and hydropathy characteristics. However, homology is not uniform between or within the proteins; nsP1, nsP2, and nsP4 contain extended domains which are highly conserved between alphaviruses, while the C-terminal region of nsP3 shows little conservation in sequence or length between alphaviruses. An untranslated "junction" region of 44 nucleotides (for RRV NB5092) or 47 nucleotides (for RRV T48) separates the nonstructural and structural protein coding regions. The structural proteins (capsid-E3-E2-6K-E1) are translated from an open reading frame of 3762 nucleotides which is followed by a 3'-untranslated region of approximately 348 nucleotides (for RRV NB5092) or 524 nucleotides (for RRV T48). Excluding deletions and insertions, the genomes of RRV NB5092 and RRV T48 differ at 284 nucleotides, representing a sequence divergence of 2.38%. Sequence deletions or insertions were found only in the noncoding regions and include a 173-nucleotide deletion in the 3'-untranslated region of RRV NB5092, compared with RRV T48. In the coding regions, most of the nucleotide differences are silent; there are 36 amino acid differences in the nonstructural proteins and 12 in the structural proteins. The distribution of amino acid differences between the two RRV strains correlates with the location of domains which are poorly conserved in sequence between alphaviruses. The possible role of amino acid differences in envelope glycoproteins E1 and E2 in determining the different antigenic and biological properties of RRV NB5092 and RRV T48 is discussed.

Alphavirus↗
PubMed1982

Co-expression of mouse and rat haemoglobins in interspecific hybrids of mouse and at erythroleukemia cells.

A modified technique for cell fusion with lysolecithin-lipid emulsions was used to generate hybrid erythroleukemia cell lines from Friend leukemia mouse cells (FLC) and chemically transformed rat erythroleukemia cells. Chromosome analysis of the hybrid cells showed the presence of both parental genomes even after long culture periods. The hybrids were still able to undergo erythroid differentiation after dimethylsulphoxide (DMSO) stimulation. Analysis of the globin chains from the DMSO-stimulated cells showed that both the rat and the mouse erythroid phenotypes were expressed. This demonstrates the compatibility of the regulatory genetic elements for the control of erythroid differentiation in cell hybrids of erythroleukemic populations from different species.

Animals↗
PubMed1986

Ontogenetic development of lipopolysaccharide-reactive B cells against bromelain-treated mouse erythrocytes in mouse lymphoid tissues.

In lymphoid tissues of mice, there exist LPS-reactive B cells which can differentiate to IgM-secreting plaque-forming cells (IgM-PFC) and PFC secreting antibodies against bromelain-treated mouse erythrocytes (BrMRBC) by LPS activation. In this study, four groups of LPS-reactive B cells in spleen, peritoneal cavity and mesenteric lymph nodes from 2- and 10-week-old mice were compared and enumerated as precursors of IgM-PFC and anti-BrMRBC PFC on days 1 and 2 after LPS activation in quantitative culture conditions. The induction of each of four PFC responses in peritoneal cells was sensitive to LPS and anti-mouse IgM antibodies as much as the induction of the respective PFC response in spleen cells. The ratios of four groups of PFC to each other were different among three tissues and between two ages. These findings support the view that the four groups of LPS-reactive B cells in each tissue are mostly in distinct subpopulations of B cells from each other, and the respective groups of different lymphoid tissues at different ages belong to the same subpopulation.

Aging↗
PubMed1983

Improvements in obtaining and characterizing mouse cerebrospinal fluid. Application to mouse hepatitis virus-induced encephalomyelitis.

This report describes advances in techniques for analyzing cellular and humoral immune components in the cerebrospinal fluid (CSF) of the mouse that are applicable to other laboratory animals. CSF studies undertaken during experimental infection of mice with JHM strain virus (JHMV) of mouse hepatitis virus are presented. A critical pitfall which can lead to erroneous or invalid results is contamination of the CSF by even minute quantities of blood. Means of avoiding this contamination are attention to anatomical reference points, the use of a micropipet, and prior intracardiac perfusion of animals with phosphate-buffered saline. Cells in the CSF were typed as either B, T, polymorphonuclear, or mononuclear cells by the combination of a microcytotoxicity assay and histologic stains. A radioimmunoassay (RIA) allowed quantification of antibodies to JHMV in the CSF and indicated the presence of intrathecal synthesis of antibody in chronically infected mice. The combined use of these sensitive methods makes possible CSF analysis in individual mice rather than in pooled groups.

Animals↗
PubMed1990

Comparative sensitivity of infectivity assay and mouse antibody production (MAP) test for detection of mouse thymic virus (MTLV).

Mouse thymic virus (murid herpesvirus-3; MTLV) is a naturally occurring T lymphotropic herpesvirus of mice. We compared the sensitivity of infectivity assay, which tests for induction of thymic necrosis in newborn mice, and an enzyme immunoassay (ELISA)-based modified mouse antibody production (MAP) test. Infection in adult mice was verified by infectivity assay of salivary glands. Approximately ten times as much virus was required to infect adult mice as newborns. No adults became infected at the lowest dose (just below 1 ID50 by thymic necrosis in newborn mice); a dose tenfold higher resulted in both infection and seroconversion in three out of five mice. At higher doses tested (up to 8000 ID50), all mice became infected and seroconverted. Infected mice shed virus and remained seropositive for at least six months. False positives were observed in ELISA; these could be eliminated by adding control (uninfected) thymic homogenate (1/10 volume of a 10% homogenate) to the sample diluent. These data suggest that the MAP assay can be a reliable and sensitive indicator of infection in adult mice undergoing primary infection. Although slightly less sensitive than infectivity assay for detecting MTLV, the MAP test is likely to detect most samples containing virus. It is recommended that several mice be used for each sample and that all positive serum samples be retested with control thymic homogenate for increased reliability.

Animals↗