Masahiko Nishimura

Chromosomal mapping of the host resistance locus to rodent malaria (Plasmodium yoelii) infection in mice

Abstract. The disease outcome in malaria caused by the protozoan parasite Plasmodium is influenced by host genetic factors. To identify host genes conferring resistance to infection with the malaria parasite, we undertook chromosomal mapping using a whole-genome scanning approach in cross-bred mice. NC/Jic mice all died with high parasitemia within 8xa0days of infection with 1×105 parasitized erythrocytes. In contrast, 129/SvJ mice all completely excluded malaria parasites from the circulation and remained alive 21xa0days after infection. We performed linkage analysis in backcross [(NC/Jic×129/SvJ)×NC/Jic] mice. The Pymr (Plasmodium yoelii malaria resistance) locus was mapped to the telomeric portion of mouse Chromosome (Chr)xa09. This locus controls host survival and parasitemia after infection. The Char1 locus (P. chabaudi resistance locus 1), controlling host survival and peak parasitemia in P. chabaudi infection, was previously mapped to the same region. This host resistance locus mapping to Chrxa09 may represent a ubiquitous locus controlling susceptibility to rodent malaria. Elucidation of the function of this gene will provide valuable insights into the mechanism of host defense against malaria parasite infection.

Detection of a new cerebral malaria susceptibility locus, using CBA mice

Human cerebral malaria (CM) during acute Plasmodium falciparum infection is a serious neurological complication that leads to coma and death. P. berghei ANKA infection of CBA mice is a useful experimental model of CM. To identify host susceptibility loci, we performed chromosomal mapping in crossbred populations of both CM-susceptible CBA and CM-resistant DBA/2 mice. One significant region for a CM-susceptible locus in CBA mice was mapped to H2 region on Chromosome 17, tentatively designated cmsc. cmsc was mapped to a different chromosomal region from that previously reported in the C57BL/6 mouse model of CM. It is possible that different loci contribute to CM in CBA and C57BL/6 mouse strains. Comparison of the function of CM susceptibility loci between CBA and C57BL/6 mice could have important implications for the study of the complex pathogenesis of CM in humans.

Mapping quantitative trait loci for circadian behavioral rhythms in SMXA recombinant inbred strains.

SM/J and A/J inbred strain of mice have different characteristics in circadian behaviors such as free-running period (τ), phase relationship (ψ) between light-dark cycles and activity rhythms, and amount of wheel-running activity. To determine the genes which affect these behaviors, a quantitative trait locus (QTL) analysis using SMXA recombinant inbred strains derived from SM/J and A/J mice was performed. Concerning τ, two regions on chromosomes (Chrs) 7 and 18 surpassed the genome-wide suggestive level. As for ψ, one suggestive QTL was detected on Chr 7. The QTLs which affect daily activity counts under light-dark cycles and constant darkness were mapped to the same chromosomal regions on Chrs 1 and 17, respectively. The provisional QTLs detected in the present study might be useful for understanding the complex mechanism regulating circadian behaviors.

Major quantitative trait locus on chromosome 2 for glucose tolerance in diabetic SMXA-5 mouse established from non-diabetic SM/J and A/J strains

Aims/hypothesisThe SMXA-5 mouse is one of the SMXA recombinant inbred substrains established from the non-diabetic SM/J and A/J strains, and is a model for polygenic type 2 diabetes, characterised by moderately impaired glucose tolerance and hyperinsulinaemia. These diabetic traits are worsened by feeding a high-fat diet. The aim of this study was to dissect the diabetogenic loci in the A/J regions of the SMXA-5 genome that contribute to diabetes-related traits.Materials and methodsWe analysed the quantitative trait loci (QTL) for diabetes-related traits and obesity in (SM/J×SMXA-5)F2 intercross mice fed a high-fat diet. To verify the function of the responsible locus that was mapped in the present study, we constructed a congenic strain and characterised its diabetes-related traits.ResultsA major QTL for glucose tolerance, free-fed blood glucose concentration and BMI was mapped on chromosome 2. This locus existed near D2Mit15, with the highest logarithm of the odds score (12.6) for glucose concentration at 120xa0min in a glucose tolerance test, and was designated T2dm2sa. The diabetogenic allele of T2dm2sa originated in the A/J strain. SM.A-T2dm2sa, a congenic strain that introgressed the T2dm2sa region of A/J genome into SM/J, exhibited overt impaired glucose tolerance and hyperinsulinaemia.Conclusions/interpretationThe development of impaired glucose tolerance in SM.A-T2dm2sa mice confirmed the results of QTL analysis for diabetes-related traits in F2 intercross mice. The present results suggest that there are latent diabetogenic loci in the genomes of non-diabetic A/J and SM/J mice, and that the coexistence of these loci, including T2dm2sa, causes impaired glucose tolerance in SMXA-5 and SM.A-T2dm2sa mice.

SMXA-5 Mouse as a Diabetic Model Susceptible to Feeding a High-fat Diet

The SMXA-5 strain, a new mouse model for type 2 diabetes, is a recombinant inbred strain derived from non-diabetic SM/J and A/J strains. As dietary fat is a key component in the development of diabetes, we compared the glucose tolerance and diabetes-related traits among the SMXA-5, SM/J, and A/J strains while feeding a high-fat diet for 10 weeks. SMXA-5 fed on a high-fat diet showed an increased serum insulin concentration. Judging from the hyperinsulinemia in SMXA-5, this strain showed insulin resistance, an inability of peripheral tissues to respond to insulin, which was strengthened by feeding with a high-fat diet. When fed on a high-fat diet for 5 weeks, the SMXA-5 mice showed severely impaired glucose tolerance. On the other hand, SM/J showed mildly impaired glucose tolerance, even when fed on a high-fat diet for 10 weeks. These results indicate that SMXA-5 would be available for use as a diabetic model susceptible to a high-fat diet.

Fas has a role in cerebral malaria, but not in proliferation or exclusion of the murine parasite in mice

We examined the susceptibility of murine Fas-deficient mutants to malaria infection in order to investigate the role of Fas in an experimental murine model of cerebral malaria (CM). We infected mice of B6 and CBA wild-type and mutant backgrounds with Plasmodium berghei ANKA. The incidence of CM in the mutant mice (B6-lpr, CBA-lprcg) was decreased by about 50% compared with wild-type control strains at 2 weeks after infection. We did not observe significant differences of parasitemia during a murine malaria infection with nonlethal Plasmodium yoelii 17XNL between wild-type and lymphoproliferative (lpr) mutant mice of C3H and MRL genetic backgrounds, although B6-lpr mice exhibited significantly higher parasitemia than did B6 mice 12 to 18 days after infection. These results suggest Fas has a possible role in CM but may not play a major role in the proliferation or exclusion of a murine malaria parasite in a nonlethal infection.

Difference in average survival between F344/Du and F344/N rats is not due to genetic contamination.

The average survival was found to be different between substrains of rats, F344/Du and F344/N. One of the causes of this difference could have been genetic contamination by crossbreeding with other strain(s) after the divergence of substrains. These two strains were examined by PCR-SSLP (simple sequence length polymorphism) procedure using microsatellite (Mit) markers to identify the contamination. All 288 Mit markers distributed throughout the genome exhibited no size polymorphisms between these two strains. This indicated that there was no genetic contamination between these two strains. The phenotypic difference in average survival, thus, is not due to genetic contamination.

Deletion in the beige gene of the beige rat owing to recombination between LINE1s

Abstract. We have determined the molecular genetic basis of the rat beige mutant, a model for human Chediak-Higashi syndrome. Deletion of a 578-bp sequence, which led to a frame shift and a presumably non-functional truncated BEIGE protein, was identified in beige cDNA. The beige rat had a deletion of about 20 kb of genomic DNA, including three exons, which constitute the deleted 578-bp cDNA fragment. LINE1s (Long Interspersed Nucleolar Element 1) were identified at the site of the deletion. Consensus recognition sequences for DNA topoisomerase I were clustered at the putative deletion junction sites in LINE1s. We conclude that the deletion in the beige gene mediated by recombination between LINE1s is the causative mutation in the beige rat. The recombination might have been induced by DNA topoisomerase I and the extensive sequence homology between LINE1s.

Chromosomal mapping of host susceptibility loci to Angiostrongylus costaricensis nematode infection in mice.

Abstract.Angiostrongylus costaricensis is a nematode found mainly as a rodent parasite. Laboratory mice were experimentally infected with this parasite. It is known that there is great variability in mortality among inbred mouse strains after infection with this nematode. The survival rate at 5xa0weeks after infection of A/J mice was 90.5%, whereas that of SM/J mice was only 33.3%, with severe anemia and decreased body weight about 3xa0weeks after infection. To identify host susceptibility genes for infection with this nematode, we undertook chromosomal mapping by a whole-genome scanning approach in (A/J×SM/J)F2 mice. We mapped a host susceptibility locus (here designated Acsns, for Angiostrongylus costaricensis nematode susceptibility locus) to the telomeric portion of Chromosome 19 (peak LOD=4.35). We also identified two loci on Chrxa013 and Chrxa017 that have epistatic effects on host survival. This is the first report on host susceptibility loci for helminth infection mapped by whole-genome scanning.

Establishment of consomic strains derived from A/J and SM/J mice for genetic analysis of complex traits.

Consomic strains, in which one chromosome is derived from a donor strain and the other chromosomes are derived from the recipient strain, provide a powerful tool for the dissection of complex genetic traits. In this study we established ten consomic strains (A-2SM, A-6SM, A-11SM, A-12SM, A-13SM, A-15SM, A-17SM, A-18SM, A-19SM, A-YSM) using the SM/J strain as the donor and the A/J strain as the recipient; these are the parental strains of a set of SMXA recombinant inbred (RI) strains that we had developed previously. We analyzed body weights and blood lipid levels in the consomic and parental strains. The mean values for each trait showed a continuous range of variation in the consomic strains suggesting that they are controlled by multiple genes. We previously identified suggestive QTLs for body weight on chromosome 6 in SMXA RI strains and (SM/Jxa0×xa0A/J)F2 mice. The observation that the A-6SM consomic strain had a significantly lower mean body weight than the A/J strain supports the presence of this QTL on chromosome 6. Similarly, the higher blood triglyceride level in the A-11SM strain shows the existence of a previously mapped QTL on chromosome 11, and the A-12SM strain provides evidence of a QTL for blood total cholesterol level on chromosome 12. These consomic strains, along with the previously developed set of SMXA RI strains from A/J and SM/J mice, offer an invaluable and powerful resource for the analysis of complex genetic traits in mice.