Transgenic major histocompatibility complex class I antigen expressed in mouse trophoblast affects maternal immature B cells.

We have produced transgenic mice using the mouse placental lactogen type II promoter to force and restrict the expression of the mouse major histocompatibility complex (MHC) class I molecule, H-2K(b), to the placenta. We show that the transgenic MHC antigen H-2K(b) is expressed exclusively in trophoblast giant cells from Day 10.5 until the end of gestation. This expression affects neither the fetal development nor the maternal tolerance to the fetus in histoincompatible mothers. We have used the 3.83 B cell receptor (BcR) transgenic mouse line to follow the fate of H-2K(b)-specific maternal B cells in mothers bearing H-2K(b)-positive placentas. Our results suggest that transgenic H-2K(b) molecules on trophoblast giant cells are recognized by 3.83 BcR-transgenic B cells in the bone marrow of pregnant females. This antigen recognition triggers the deletion of a bone marrow B cell subpopulation, including immature and transitional B cells. Their percentage decreases during the second half of gestation and is down to 8% on Day 17.5, compared to 22% in the (3.83 Tg female x Fvb) control group. This deletion might contribute to the process of maternal tolerance of the conceptus.

Comparative diagnoses of twin zygosity by SSLP variant analysis, questionnaire, and dermatoglyphic analysis.

Zygosity diagnosis has been performed in 79 pairs of twins using three methods. Simple sequence repeat length polymorphism (SSLP) analysis allows an efficient classification (MZ or DZ) with only a few markers following a simplified technique of extraction and amplification. A method based on a full questionnaire completed by parents about twin similarity correctly classifies 97.46% of the pairs; 92.41% are correctly classified using only four questions as suggested by logistic regression analysis. The third method, using dermatoglyphic analyses, correctly classifies 86.76% of pairs. To lower the cost of DNA diagnosis we stress the possibility of limiting its use to pairs with scores in the overlap area between MZ and DZ twins with a validated questionnaire.

Linkage between brain serotonin concentration and the sex-specific part of the Y-chromosome in mice.

The implication of the sex-specific part of the Y-chromosome (YS-SP) on brain serotonin (5-HT) level was investigated using congenic strains for this chromosomal region. The 5-HT level, which was higher in the NZB than in the CBA/H strain of mice, was depleted by the transfer of the YS-SP from NZB on CBA/H whereas the transfer of the YS-SP from CBA/H on NZB had no effect. The variations of 5-HT levels were not correlated with plasma testosterone concentration which is also dependent of the YS-SP.

Long-term effects of embryo freezing in mice.

Embryo cryopreservation does not induce clear-cut anomalies at detectable rates, but several mechanisms exist for nonlethal damage during the freeze-thaw process, and the risk of moderate or delayed consequences has not been extensively investigated. In a long-term study including senescence, we compared cryopreserved and control mice for several quantitative traits. Significant differences were seen in morphophysiological and behavioral features, some of them appearing in elderly subjects. Thus, apart from its immediate toxicity, embryo cryopreservation, without being severely detrimental, may have delayed effects. These results, consistent with other findings, question the neutrality of artificial reproductive technologies and draw attention to the preimplantation stages in developmental toxicology.

Alleles of the microsomal steroid sulfatase gene (Sts) in the pseudoautosomal region of the heterosomes of the mouse.

The gene (Sts) for microsomal steroid sulfatase (STS-EC 3.1.6.2) is located at the distal end of the pseudoautosomal region of the mouse heterosomes. A new improved method was developed to measure the activity of this enzyme in four inbred strains of laboratory mice, their respective congenics for the non pseudoautosomal region of the Y chromosome and their F1s were employed. The method was shown to be highly reliable (.95). No sexual dimorphism was shown but a polymorphism for the enzymatic activity, corresponding to three functional allelic forms was found. The results presented here are compatible with the location of the Sts gene on the X-Y pseudoautosomal region. The underdominance that appears in the F1s, in one set of strains and the difference between reciprocal F1s in the other set must be interpreted cautiously since it is the enzymatic activity which is only measured and not the protein itself. A complex mechanism, involving the gene responsible for the enzyme located on the pseudoautosomal region and autosomal locus or loci which regulate(s) its activity, is suggested as being responsible for the results obtained with the F1s.

Co-segregation of intermale aggression with the pseudoautosomal region of the Y chromosome in mice.

The sexual dimorphism of aggression has led to a search for its Y chromosomal correlates. We have previously confirmed that initiation of attack behavior against a conspecific male is Y-dependent in two strains of laboratory mice (NZB and CBA/H). We provide evidence that the non-pseudoautosomal region of the Y is not involved and that only the pseudoautosomal region of the Y is correlated with initiation of attack behavior. The autosomal correlates also contribute to this behavior in an additive or interactive manner with the pseudoautosomal correlates.

Hippocampal mossy fiber changes in mice transgenic for the human copper-zinc superoxide dismutase gene.

The copper-zinc superoxide dismutase (SOD-1) gene, located on chromosome 21 and triplicated in Down's syndrome (DS), is suspected to be involved in the neuropathology observed in Alzheimer's disease (AD), DS and physiological aging. In order to explore the effect of an overproduction of SOD-1 in the mouse hippocampus, we investigated the Timm-stained mossy fiber (MF) innervation in the hippocampus of transgenic mice for the human SOD-1 gene (hSOD-1 mice). The results showed a decrease of the MF projection area in the hSOD-1 mice overexpressing the SOD-1 protein. These findings suggest that free radicals could play a role in this particular synaptic loss.

Mandible shape analysis in a testicular feminization (Tfm) strain of mice.

The role of androgens on the sexual dimorphism of mandible shape was investigated in mice carrying the X-linked gene for testicular feminization (Tfm), which is known to determine a profound insensitivity to testosterone and is associated with a severe reduction in androgen receptor levels in Tfm/Y males. Mandible shape analysis in an inbred strain of mice segregating for the Ta (tabby) and Tfm mutations showed that the sexual dimorphism observed between +Ta/+Ta females and +Ta/Y males almost disappeared between Tfm+/+Ta females and Tfm+/Y males. In addition, a canonical discriminant analysis showed that these two closely related classes, Tfm+/+Ta and Tfm+/Y, are readily differentiated from both the +Ta/+Ta and +Ta/Y classes. These results suggest that androgens are involved in the mandible shape sexual dimorphism and play a role in mandibular development in both males and females.

Genetic analysis of isolation-induced aggression. I. Comparison between closely related inbred mouse strains.

Young adult males of the inbred mouse strains BALB/c, C57BL/6, CBA, C3H/He, and the outbred strain NMRI showed significant interstrain differences for isolation-induced aggression as measured in a standard-opponent test. Additionally, four related inbred strains of common origin (ABG, AB//Halle, ABB, AB//Jena) were studied. ABG and AB//Halle showed nearly no overlap for the trait isolation-induced aggression. Tests of histocompatibility and coat color, as well as mandible measurements, confirmed a close genetic relationship between ABG and AB//Halle. The large difference for isolation-induced aggression between these two closely related strains offers excellent opportunities to search for single gene correlates of aggression.

Intraperitoneal infection with Salmonella abortusovis is partially controlled by a gene closely linked with the Ity gene.

The aim of the present study was to determine whether the Ity gene, which controls the resistance to S. typhimurium infection in mice, also governs the resistance to S. abortusovis, a serotype specific for goat and sheep. During either i.v. or i.p. infection, BALB/c mice (Itys) were not able to control the growth of S. abortusovis and eventually died from infection. In contrast CBA (Ityr) or (C.CB)F1 (Ityr/s) mice were able to control the growth of these bacteria. Using congenic C.D2 Ityr mice, we found that the gene controlling resistance to S. abortusovis was tightly linked to the Ity gene on chromosome 1. Furthermore, in the spleen and the liver of backcross BALB/c x (CBA x BALB/c) mice, the S. abortusovis resistance phenotype cosegregated with the two alleles of the Len-1 gene, a gene tightly linked to the Ity gene. By contrast, in these backcross mice, the level of infection of the peritoneal cavity, the site of inoculation, did not correlated with the Len-1 phenotype of the animal. These results provide evidence that after i.p. inoculation the control of S. abortusovis growth in the spleen and the liver is controlled by the Ity gene, but also suggest that additional gene(s) regulate the number of bacteria at the site of inoculation.

Comparative evaluation of the immune status of congenitally athymic and euthymic rat strains bred and maintained at different institutes: 1. Euthymic rats.

We performed a comparative evaluation of the immune status, focused on the T-cell system, of euthymic rat strains in which the nude mutation had been introduced. From 10 institutes, we sampled 12 groups of euthymic rats at ages of 1 1/2-2 months and 1/2 year. We analyzed weight of body, spleen and thymus; antibody response and delayed-type hypersensitivity response to ovalbumin immunization; and (immuno)histopathology of spleen, lymph nodes, and lymphoid tissue along the gastrointestinal tract. In the spleen morphometric analysis was done of the periarteriolar lymphocyte sheath (using the antibody R73 recognizing the alpha beta-T-cell receptor) and of the red pulp (using the antibody ED2 recognizing red pulp macrophages). For almost all parameters tested, statistically significant differences between the groups (origin of the animals) were observed. A cluster analysis on the basis of body weight, spleen weight, and morphometric data of spleen did not yield clusters with a different composition among animals from individual groups. Based on the antibody response to ovalbumin, clustering revealed groups of "fast-and-high", "slow-and-low", and "intermediate" responders. The various groups differed in location within these clusters, i.e. the speed and extent of the immune response depends on the background euthymic strain. Considering the microbiological status assessed by serology, a variation was found both in post-infection state at entrance in the study, and in primo-infection associated with a rise in antibody concentrations during the study. These states showed no negative effect on anti-ovalbumin reactivity. Rather, the response in primo-infection to Rat Corona Virus, Sendai Virus, and Pneumonia Virus of Mice was the highest in animals clustered as "fast-and high" responders to ovalbumin.

Comparative evaluation of the immune status of congenitally athymic and euthymic rat strains bred and maintained at different institutes: 2. Athymic rats.

We performed a comparative evaluation of the immune status, focused on the T-cell system, in congenitally athymic rat strains. From 11 institutes around the world, we sampled 15 groups of animals at ages of 1 1/2-2 months and 1/2 year. The analysis included weight of body and spleen; antibody response and delayed-type hypersensitivity response after immunization with ovalbumin; and (immuno)histology of spleen, lymph nodes and lymphoid tissue along the gastrointestinal tract. Morphometric analysis was done for alpha beta-T-cell receptor-bearing cells in spleen tissue as a measure of the periarteriolar lymphocyte sheath; it was also done for splenic red pulp using the antibody ED2 recognizing red pulp macrophages. For almost all variables analyzed, statistically significant differences between the groups were observed. The extent of alpha beta-T-cell receptor-bearing cells in the spleen increased with age. The functioning of these cells in immunological responses can be questioned, because an immune response to ovalbumin was invariably absent. But secondary follicles with germinal centers, reflecting T-cell-dependent B-cell reactivity, were observed in lymph nodes and Peyer's patches (up to 40% and 75%, respectively, depending on the group), with a higher prevalence in older animals. A cluster analysis on the basis of body and spleen weight and composition of spleen compartments did not yield clusters with a different profile in regard to the animals' group of origin. The data presented are useful when comparing studies performed with various athymic rat strains at different institutes.

Mandible shape analysis in Y-congenic strains of mice.

In order to test for the presence of genes influencing morphology on the Y-chromosome of mice, a canonical discriminant analysis of variation in mandible shape was conducted in 4 inbred strains of mice and 4 Y-chromosome congenic partners. Genetic background-dependent differences were observed, as well as a small sex variation in some strains, but it was impossible to detect any difference related to the between-strain exchange of the Y-chromosome.

Early and transient osteopetrosis in microphthalmic MIB-rats.

Microphtalmic blanc mutation (mib/mib) displays a very mild form of osteopetrosis in rats. The autosomal recessive mib mutation shows pleiotropic expressions in homozygotes. Microphtalmia, absence of eye and skin pigmentation, retardation in the tooth eruption were observed in the mutants. Most bone abnormalities occurred in newborns. An increased radiological opacity of long bones, persistence of primitive bone in medullary cavities, reduced number of poorly differentiated osteoclasts in mandibulae, reduced number of mononuclear peritoneal cells as well as reduced number of mononuclear osteoclast precursors in peritoneal cell population were found. In 3 weeks old and in adult mutants, both bone structure and the number of mandible osteoclasts appear normal, but the number of blood monocytes, peritoneal cells and mononuclear osteoclast precursors in peritoneal cell population remain significantly lower than in the healthy littermates. These observations indicate that the early failure of osteoclast differentiation and maturation is transient in the mib/mib form of osteopetrosis.

Creation of a strain of genetically obese-hypertensive rats.

A strain of genetically obese-hypertensive rats (SHR-fa/fa) was created by transferring the fatty/fa gene of hyperlipaemic obese non-inbred rats into the genome of an SHR inbred strain by five successive crossings of SHR-fa+ brother-sister matings. SHR-fa/fa rats were heavier than their SHR littermates. They showed a severe hypertension, their systolic arterial blood pressure being higher than that previously found in genetically hypertensive rats. Their blood glucose content was not significantly different from that of their SHR littermates but their plasma triglycerides were increased by more than 500 per cent. While obesity and hypertension occurred from the 5th week following the rat's birth, the increase in blood triglycerides was manifest earlier.

Comparative study of the biosynthesis of PGE2, PGF2 alpha and TXA2 by different organs of genetically hypertensive (SHR) and obese-hypertensive (SHR-fa/fa) rats.

As an experimental model, we used 6-week-old genetically obese-hypertensive rats (SHR-fa/fa) which were obtained by transferring the fatty/fa gene of hyperlipaemic obese rats into the genome of the SHR strain: the SHR-fa/fa were bigger and more hypertensive than their SHR littermates. Studying the capacity of the hearts, kidneys, spleens, brains and lungs to synthesize PGE2, PGF2 alpha and TXA2, enabled us to show that the hearts and lungs of SHR-fa/fa synthesized more PG than those of SHR; SHR-fa/fa brains generated less icosanoids than those of SHR; the amounts of PGE2 and TXA2 produced by the kidneys are similar in SHR and in SHR-fa/fa. From the experimental data we can infer that the introduction of the fatty/fa gene into the genome of SHR does not significantly alter the capacity of the kidneys to synthesize icosanoids; the more severe hypertension in the SHR-fa/fa would result from an increase in TXA2 biosynthesis by cardiac tissue which, at the same time, synthesized more PGE2, which could be a means of defence against hypertension. Moreover this genetical manipulation inhibited the icosanoid-synthesizing capacity of the brain which thus attenuated the central nervous system activity of the animals.

Mercury-induced autoimmune glomerulonephritis: requirement for T-cells.

Mercury-induced autoimmunity in Brown-Norway rats has been shown previously to be due to polyclonal activation of B lymphocytes, requiring the presence of T lymphocytes. Autoimmunity in that strain is characterised by the appearance of an autoimmune glomerulonephritis, by the production of a host of autoantibodies, and by an increase in total serum IgE. In the present study, T-cell deprived rats were tested to assess the role of T cells in the appearance of autoimmune abnormalities in vivo. It will be shown that both BN rnu/rnu and BN 'B' rats, who have virtually no T cells, do not develop autoimmunity following HgCl2 injections. In contrast BN 'B' rats reconstituted with normal T cells, and BN rnu/+ rats, exhibit autoimmune manifestations, including autoimmune glomerulonephritis, quite similar to those observed in Brown-Norway rats. These data demonstrate that T cells are essential for mercury-induced autoimmunity to occur in Brown-Norway rats.

[Cure of osteopetrosis by injection of allogenic bone marrow into "op" rats treated with cyclosporin A]. / Guérison de l'ostéopétrose par injection de moelle osseuse allogénique chez le rat "op" traité par la cyclosporine A.

The cure of osteopetrosis by allogenic bone marrow injection has been obtained in "op" mutant rat kept under cyclosporin A treatment. This immunosuppressive agent able to prevent the rejection of transplanted cells does not impair the propriety of these cells to restore the bone resorption process in this severe osteopetrotic form.