Recognition among mice. Evidence from the use of a Y-maze differentially scented by congenic mice of different major histocompatibility types.

Previous studies of mating preference signified that mice can sense one another's major histocompatibility complex (MHC) types, probably by olfaction. This conclusion has now been substantiated by the use of a Y-maze whose two arms were differentially scented with currents of air conducted through boxes occupied by B6 (H-2b) males and by B6-H-2k congenic males. Four B6 mice, two males and two females, were successfully trained, by water deprivation and reward, to enter the arm scented by B6 or B6-H-2k males. One of the males and one of the females were trained to select the B6-scented arm; the other male and female were trained to select the B6-H-2k-scented arm. Untrained mice showed no MHC discrimination in the maze. The performance of the trained mice in distinguishing between MHC congenic homozygous F2 segregants derived from a cross of B6-H-2k with B6 was as good as their performance in distinguishing the respective inbred strains, thus essentially eliminating alternative and significant additional explanations of MHC-associated sensory discrimination. The data further indicate that chemosensory discrimination of MHC types can be entirely dissociated from sex differences and from the circumstances of mating.

Organization and evolution of D region class I genes in the mouse major histocompatibility complex.

Chromosome walking has been used to study the organization of the class I genes in the D and Qa regions of the MHC of the BALB/c mouse and in the D region of the AKR mouse. Five and eight class I genes are found in the D and Qa regions of the BALB/c mouse, respectively, while the AKR mouse contains only a single class I D region gene that has been identified by transfection as the Dk gene. Restriction map homologies and crosshybridization experiments suggest that the multiple class I genes in the D region of the BALB/c mouse have been generated by unequal crossing-over involving class I genes from the Qa region. The expanded D region of BALB/c and other H-2d haplotype mouse strains appears to be metastable, since evidence for gene contraction in the Dd region has been found in two mutant strains. Thus the D region and also the Qa region class I genes are in a dynamic state, evolving by gene expansion and contraction.

Genetic study of lymphoma induction by AKR mink cell focus-inducing virus in AKR x NFS crosses.

The mink cell focus-inducing (MCF)-247 virus, originally isolated from an AKR thymoma, is lymphomagenic in AKR mice but not in the ecotropic virus-negative NFS mouse strain. Analysis of sensitivity to lymphoma-induction by AKR-247 MCF virus in genetic hybrids between these two strains showed that F1 mice inoculated as sucklings were uniformly sensitive, whereas those inoculated as weanlings were generally resistant. In NFS backcross mice inoculated as sucklings, inheritance and expression of endogenous ecotropic virus from AKR was an essential correlate of replication of MCF virus and subsequent development of lymphoma. However, one-third of the mice expressing ecotropic virus and replicating the inoculated MCF virus did not develop lymphoma. The results suggested that an additional gene that influenced development of lymphoma may be involved, and that mice inheriting both virus-inducing loci from AKR were more susceptible than those inheriting only one. These findings indicate that the causal role of ecotropic virus infection in spontaneous thymomagenesis in AKR mice involves not only the generation of leukemogenic MCF viruses but also the establishment of permissiveness for their growth.

Evidence for the horizontal acquisition of murine AKR virogenes by recent horizontal infection of the germ line.

Several recent reports (8, 10, 11, 13) have established the biological and molecular genetic similarity between the endogenous AKV virus of strain AKR, and an N-ecotropic endogenous virus found in the genome of feral Japanese mice, Mus musculus molossinus. The similarities are so striking as to suggest a common origin of these viruses, which are present in some, but not all, inbred mouse strains. The virogenes of AKR mice may have been acquired by either: (a) common descent of AKR (and other AKV(+) strains) from a common ancestor of AKR and molossinus animals, or (b) horizontal germ line infection of the AKR strains by molossinus virus at 1;he strain's inception followed by fixation through inbreeding. The sexual descent model carries with it a prediction of relative consanguinity of the AKR strain and molossinus, whereas the horizontal infection model does not. We have examined the polymorphic allozyme (allelic isozyme) genotype of 51 nonvirus-related loci in 17 strains of mice including AKR, C58, BALB/c, Swiss, and molossinus. By comparing the composite allozyme genotype of different inbred and outbred mouse strains, the "genetic distance" statistic was derived. Genetic distance measures the degree of allelic substitution between populations and increases proportionately with the amount of time the populations have been reproductively isolated. The genetic distance computed between molossinus and AKR is large, nearly 5-10 times the distance between known related populations and strains (e.g., C57L vs. C57BL/6). Molossinus had a similarly large distance from AKV negative strains (Swiss, C57L) as it did from AKV- positive strains. Cellular DNA sequences that flank the integrated AKV provirus were analyzed by restriction enzyme digestion of liver DNA from molossinus, AKR, and additional inbred strains that express ecotropic murine leukemia virus. The integration flanks of three AKR provirus sequences, Akv-1, Akv-2, and a third uncharacterized sequence, were not evident in molossinus cell DNA, which contained at least six different proviral integration fragments. These data effectively exclude the interpretation of consanguinity of AKR and molossinus and support the notion of acquisition of the endogenous virus in AKR by horizontal infection of the molossinus virus.

The generation and specificity of cytotoxic T cells raised against syngeneic tumor cells bearing AKR/Gross murine leukemia virus antigens.

Efforts were made to generate C57BL/6 cytotoxic effector cells to a syngeneic leukemia (E{male}G2) bearing AKR/Gross virus antigens. As we were unable to induce significant cytotoxic activity by immunization with up to 10(8) irradiated E{male}G2 cells, even when cells from such primed animals were subsequently restimulated with E{male}G2 cells in vitro, C57BL/6 mice were immunized with an aliogeneic, virus-producing AKR leukemic cell line (AKR SL3). Peritoneal exudate cells and, to a lesser degree, spleen cells from these mice showed significant lytic activity toward the immunizing allogeneic tumor but not toward E{male}G2. When spleen cells were harvested from animals {approximately equal to}10 d after injection of AKR SL3 and rechallenged in vitro with either E{male}G2 or AKR.H-2(b) SL1, another tumor that displays AKR/Gross virus antigens, then a vigorous cytotoxic response against E{male}G2 and AKR. H-2(b) SL1 was obtained. Effector cells generated by AKR SL3 priming followed by in vitro stimulation with E{male}G2 or AKR.H-2(b) SL1 lysed only cells of H-2(b) haplotype which were strongly positive for the display of serologically detectable AKR/Gross virus antigens. Thus, AKR SL3 cells were not lysed nor were EL4 cells (H-2(b); but only weakly positive for gp70). Cells not bearing the MuLV antigens tested for, such as P815 mastocytoma cells and spleen cell "blasts" from C57BL/6 and CBA (H-2(k)) mice, were also insusceptible to attack. The cytotoxic effector cells induced bore Thy 1.2 alloantigen and were of the Lyt 1+2+ phenotype. Collectively, these findings are consistent with the conclusion that the cytotoxic T cells raised against E{male}G2 are directed against AKR/Gross virus-associated antigens and are H-2 restricted. It will be of interest to determine the relevance of such effector cells to the known resistance of the C57BL/6 mouse to AKR/Gross virus-induced leukemia.

Qat-4 and Qat-5, new murine T-cell antigens governed by the Tla region and identified by monoclonal antibodies.

Two new lymphocyte antigens, provisionally designated Qat-4 and Qat-5 have been identified with two different hybridoma-derived, monoclonal AKR antiC57BL/6 antibodies. These antigens are governed by genes located to the right (distal) end of the H-2 complex, within the Qa-2,3 region. Qat-4 and Qat-5 antigens which do not seem to be identical with Qa-2,3 or TL antigens are absent from Ig/ lymphocytes and thymocytes. They are only present on a fraction of peripheral T cells. Thus, Qat-4 is expressed on 70%, and Qat-5 on 30% of splenic and lymph node T cells, Qat-4 is also found on the majority of Ig- cells from athymic nude mice. These findings illustrate the complexity of the chromosome segment between the H-2D and Tla loci and they emphasize the role of major histocompatibility complex-associated genes for the differentiation of T cells into different subpopulations with possibly distinct immunologic functions.

A mouse gene on chromosome 5 that restricts infectivity of mink cell focus-forming recombinant murine leukemia viruses.

DBA/1, DBA/2, CBA/N, and CBA/Ca mice carry a gene which specifically restricts infectivity of mink cell focus-forming (MCF) murine leukemia viruses. The gene, designated Rmcfr, is dominant or semidominant and maps to chromosome 5; it is closely linked to the morphologic marker gene Hm. The Rmcf gene may be of much use as a means of determining the role of MCF viruses in various forms of leukemogenesis.

Genes affecting mink cell focus-inducing (MCF) murine leukemia virus infection and spontaneous lymphoma in AKR F1 hybrids.

An assessment of the importance of mink cell focus-inducing (MCF)-type recombinant murine leukemia viruses in spontaneous thymic lymphomagenesis and of the genetic factors affecting its occurrence was carried out with F1 hybrids between AKR and various other inbred strains. There was generally close agreement between the frequency of detection of MCF virus, of thymocyte antigenic amplification in the preleukemic period, and of spontaneous lymphoma. Also, hybrid combinations with moderate to high spontaneous lymphoma were uniformly susceptible to lymphoma induction by neonatal inoculation of MCF 247 virus, while lower leukemic hybrids were at least partially resistant to the induced disease. At least four resistance genes can be identified as affecting the disease in the various hybrids: Fv-1, Rmcf, an unidentified gene carried by the C57 series of mice and SJL, and an unidentified minor gene carried by several other strains.

Thymic lymphoma induction by the AKT8 murine retrovirus.

The directly transforming murine retrovirus, AKT8, was isolated from a spontaneous AKR thymoma and carries the cell-derived viral oncogene, akt. We have now shown that this virus produces thymic lymphomas after inoculation of susceptible mouse strains. The presence of the AKT8 genome in the DNA of the virus-induced tumors was demonstrated by Southern blotting using an akt-specific probe. These results establish the in vivo pathogenicity of the AKT8 virus and its akt oncogene, and imply a potential role for the cellular akt proto-oncogene in tumor development.

Anergy for delayed-type hypersensitivity in preleukemic AKR mice.

A singular anergy for delayed-type hypersensitivity (DTH) in preleukemic AKR mice was discovered. This total anergy for DTH against allogeneic cells, which developed in the AKR mice by 4 to 5 months of age, was not due to an artifact of route of sensitization or of other assay parameters and was not found in other strains sharing H-2 or other genetic background. The mice had an intact capacity to be stimulated in mixed lymphocyte culture to produce cytotoxic effector cells. Although the relationship to lymphoma was not directly addressed in these experiments, the genetic and temporal characteristics of this anergy suggest a biologically important relationship to the preleukemic state.

Clonal nature of mink cell focus-inducing virus-induced AKR leukemia: studies with X-chromosome inactivation cellular mosaicism.

The number of cells from which murine thymic leukemias (thymomas) develop after neonatal injection with a mink cell focus-inducing recombinant virus was studied in AKR mice heterozygous at the X-linked phosphoglycerate kinase (PGK) locus. Because only one of the two X-chromosomes is active in XX somatic cells, thymic leukemias that are clonal should display either type A or type B PGK but not both, whereas those with a multicellular origin may exhibit both enzymes. In 23 of 25 animals studied, thymomas expressed exclusively (11 animals) or predominantly (12 animals) a single enzyme in contrast to normal tissue which expressed both enzyme types in approximately equal ratios. In the 12 thymomas expressing a minor enzyme component, the predominant enzyme in the original tumor always predominated in the thymomas arising in animals transplanted with the original tumors, indicating that this minor PGK component was not contributed by malignant cells. The results indicate that the great majority of recombinant virus-induced AKR leukemias are clonal.

Genetic study of lymphoma induction by Friend murine leukemia virus in crosses involving AKR mice.

The AKR mouse strain is uniquely susceptible to thymic lymphoma. Friend murine leukemia virus (F-MuLV) induced thymic lymphoma in this strain after a median latency of 3-4 months, whereas in several other mouse strains F-MuLV caused erythroblastosis. In the investigation of genetic determinants of the AKR predisposition to thymic lymphoma, series of congenic mouse strains and progeny of crosses involving AKR mice were inoculated with F-MuLV; these mice were followed for the development of lymphoma. The results suggested that AKR genes on chromosomes 7 and 15, other than inherited ecotropic virus, are involved in the predisposition to rapid onset of thymic lymphoma after inoculation with F-MuLV.

Lymphomagenesis in AKR.Fv-1b congenic mice.

In the AKR.Fv-1b congenic strain the Fv-1n allele of the AKR/J mice was substituted with the Fv-1b allele, thereby limiting viral replication and spread of the endogenous N-tropic murine leukemia virus. As a result of this genetic change AKR.Fv-1b mice develop a low spontaneous incidence (7%) of T-cell lymphomas and about 28% of Ly-1+ B-cell lymphomas are observed in old mice. Characteristic changes in thymus subpopulations of AKR/J mice (related to the formation of the dual tropic mink cell focus inducing (MCF) type virus in the thymus) were not observed in the thymus of AKR.Fv-1b mice. In contrast to the low susceptibility to spontaneous T-cell lymphoma development, these mice were highly sensitive to fractionated irradiation or to radiation leukemia virus (a mixture of N- and B-tropic viruses) induced T-cell lymphoma. Potential lymphoma cells (that would ultimately develop into Ly-1+ B-cell lymphomas) were demonstrated in bone marrow and spleens of 16-24-month-old mice. Analysis of the Ly-1+ IgM+ B-cell population in spleens of 18-month-old mice revealed a significant increase in this population (35% versus 2% in young spleens). The spontaneous Ly-1+ B-cell lymphoma incidence could be enhanced (up to 77%) by in vivo administration of anti-CD8 monoclonal antibody or IL-4 to 18-month-old mice. Virological analysis of T/B-cell lymphomas for class I MCF viruses indicated that Class I MCF development was tightly correlated with T-lymphoma development (except radiation induced tumors that showed no MCF provirus involvement). In contrast, Ly-1+ B-cell lymphoma development was independent of Class I MCF pathogenic virus involvement.

Role of the AKR gene locus AKv-1 in susceptibility to chemical induction of thymic lymphomas.

Various strains of mice demonstrate widely differing susceptibility to chemical induction of thymic lymphomas, in both timing and incidence. In AKR mice tumors appear very early and at high incidence after a single dose of N-methyl-N-nitrosourea, while in other strains they appear later and at lower incidences. In an attempt to determine the potential role of AKR ecotropic murine leukemia virus loci in this process, congenic mice of NFS/N background, into which the highly productive ecotropic murine leukemia virus loci AKv-1 or AKv-2 has been transferred, were challenged with N-methyl-N-nitrosourea. Although they had a lower incidence of thymic lymphomas than did the parental donor AKR, the NS.AKv-1 mice had a tumor incidence twice that of NFS/N or NS.AKv-2. However, no difference in timing was noted, and these three strains demonstrated tumor appearance much later than that of AKR/N. It is suggested that the presence of the AKv-1 loci, or a gene of the closely associated genomic region, increases the number of target cells that are susceptible to N-methyl-N-nitrosourea.

Comparison of sister chromatid exchanges in spleen and thymus lymphocytes from AKR, C57BL/6N x DBA/2J F1, and CBA mice following in vivo exposure to N-methyl-N-nitrosourea.

The induction of sister chromatid exchange (SCE) by N-methyl-N-nitrosourea (MNU) was studied in spleen and thymus lymphocytes from AKR mice, which are highly susceptible to MNU-produced thymomas, CBA mice, which are much less sensitive to induction of thymomas by MNU, and C57BL/6N x DBA/2J F1 mice. MNU produced dose-related increases in SCE in concanavalin A (Con A)- and lipopolysaccharide-stimulated spleen lymphocytes and Con A-stimulated thymus lymphocytes from each mouse strain at 1 and 24 h posttreatment. MNU-induced SCE were generally higher in Con A-stimulated spleen lymphocytes compared to lipopolysaccharide-stimulated spleen lymphocytes and Con A-stimulated thymus lymphocytes from each mouse strain. On the whole, MNU-produced SCE were lower in AKR and CBA spleens than in C57BL/6N x DBA/2J F1 spleens. In addition, MNU-induced SCE levels in thymus lymphocytes from all three strains of mice were, for the most part, similar. These results indicate that if differences in MNU-induced genotoxicity in AKR, CBA, and C57BL/6N x DBA/2J F1 thymus lymphocytes exist, these differences cannot be ascertained by use of the in vivo/in vitro SCE assay.

Autosomal dominant mouse cataract (Lop-10). Consistent differences of expression in heterozygotes.

The clinical and histologic features are reported of an autosomal dominant mouse cataract that was first observed as a new mutation in a cross between BALB/cJ and AKR/J. In the homozygous state, the eyes were microphthalmic, and a dense white cataract was present when the eyes opened at day 12. Histologic changes were apparent from birth and as early as 18 days' gestation. Liquefaction started by day 4, and herniation of lens contents posteriorly was seen at day 11. Heterozygous mice had variable expression depending both on their genetic background and age. When the single gene was expressed fully, the cataract appeared as a fetal nuclear white opacity; partial expression gave a nuclear haze to snowflake nuclear opacities. Lop-10 appeared to be an excellent model for studying variable expression of a dominant gene.

Strain differences in amphetamine sensitivity in mice. I. A diallel analysis of open field activity.

Experiments are reported which show that 1 mg/kg of d-methylamphetamine HCl induced hyperactivity in pigmented strains (C57BR, C57BL/6, and SEC) and hypoactivity or no change in albino strains (BALB/c, A, and AKR) of mice. In F1 hybrids, the B6 genotype was partially dominant over BR and C, and BR over C. In animals back-crossed to C parents widespread distributions with two peaks were obtained in control experiments, and amphetamine induced hyperactivity in 38% of the albino population, and hypoactivity or no significant change in 45% of the pigmented one. This genetic study indicates that genes influencing locomotor activity are independent from those influencing amphetamine sensitivity. From results obtained in back-crosses and C57BL/6-c2J mice, the albino gene does not seem to be involved in the hypoactive effect of amphetamine.

Genetic control of apomorphine-induced climbing behavior in two inbred mouse strains.

Over a wide dose range (1-32 mg/kg), apomorphine-induced climbing behavior was significantly greater in the AKR/J than in the DBA/2J inbred mouse strain. A similar difference was found when apomorphine-induced stereotypy was examined. A cross-fostering study demonstrated that the strain difference in apomorphine-induced climbing behavior did not result from differences in post-natal rearing environment. After apomorphine administration, brain levels of the drug were similar in the two strains indicating that the difference in behavioral response to apomorphine in the two strains was not due to differences in metabolism or distribution of the drug. The climbing response to apomorphine was examined in the F1 cross of the two strains (AKD2F1/J) and in 10 AKXD recombinant inbred strains. Results suggested that the trait was partially dominant and not X-linked; furthermore, a few and possibly one locus was responsible for the differences in apomorphine-induced climbing behavior observed in the AKR/J and DBA/2J mice.

Interstrain variation in murine aerobic capacity.

PURPOSE: The contribution of genetic factors to aerobic capacity is unknown. The purpose of this study was to measure maximal aerobic performance among inbred strains of mice to provide basic heritability estimates. METHODS: Eight female mice, 8 to 10 wk old, in 10 inbred strains (A/J, AKR/J, Balb/cJ, C(3)H/HeJ, C57Bl/6J, C57L/J, C(3)Heb/FeJ, CBA/J, DBA/2J, and SWR/J) were run on a treadmill until exhaustion. The protocol started at 22 m.min(-1) and increased in speed approximately 6 m.min(-1) every 4 min. After 4 min at 42.4 m.min(-1), the grade was increased 2% every 4 min thereafter until the mouse could not run off of the shock grid (150 V; 1.5 mA). RESULTS: There were significant differences between inbred strains in maximal duration of exercise accomplished (P < 0.0001). The order of strain-specific exercise duration was Balb/cJ > SWR/J > CBA/J > C57L/J > C3H/HeJ > C3Heb/FeJ > C57Bl/6J > AKR/J > DBA/2J > A/J. Two measures of heritability in the broad sense, intraclass correlation (0.73), and the coefficient of genetic determination (0.58) were both significant. CONCLUSION: These data indicate that there is a strong genetic contribution to aerobic capacity in mice.

Anomalous (preduodenal) portal vein: autosomal recessive mutation in AKR/J mice.

BACKGROUND AND PURPOSE: Anomalous (preduodenal) portal vein was found in AKR/J mice. It is a rare congenital malformation in humans, and to the authors' knowledge, has never been reported in laboratory animals. Morphology, clinical signs of disease, and heritability of this anomaly were examined. METHODS: Fifty-three strains of inbred mice (6,026 mice) in our mouse colony were examined for preduodenal portal vein and its association with clinical signs of disease (vomiting or abdominal pain) and other anomalies. Heritability also was tested by use of cross-backcross matings of AKR/J mice with clinically normal PT mice. RESULTS: The portal vein was found at the ventral side of the duodenum in most (98%) AKR/J mice, whereas it ran at the dorsal side of the duodenum in 52 other inbred mouse strains in our mouse colony. Clinical signs of disease and other congenital anomalies were not detected in this strain of mice, although position has a high association with other congenital anomalies in humans. Heritability testing of this anomaly in AKR/J mice indicated single autosomal recessive inheritance. CONCLUSIONS: Preduodenal portal vein found in AKR/J mice is a single autosomal recessive mutation, but is not associated with clinical signs of disease and other congenital malformations.