Interstrain differences in the maturation of congenital reflexes in 101/HY and CBA mice in early postnatal ontogeny]

We have identified interstrain differences in the rate of formation of certain reflexes and parameters of physical development in CBA/LacSto and 101/HY mice. We have shown that in young 101/HY mice, the maturation of reflexes reflecting the development of the vestibular, neurosensory, and neuromuscular systems, such as surface righting, bar holding, negative geotaxis, and auditory startle response, takes place later during postnatal life. Opening of eyes and auditory canals in mice of all studied groups takes place at the same time. In CBA females, maturation of cliff avoidance reflex occurs later than in 101/HY females; hair also appeared later. Body weight of young 101/HY mice of both sexes is significantly higher during the period of postnatal development from day 2 to day 20 than in the CBA strain. However, the relative brain weight was lower in the 101/HY strain. CBA males had a higher brain weight and also showed a faster rate of formation of inborn reflexes. We discuss possible factors underlying the observed difficulties in the rate of formation of reflexes in 101/HY and CBA young mice in relation to general background information about their genetic and neurobehavioral features. The results provide evidence that these differences are genetically determined; the rate of reflex formation does not depend on the overall physical development of mice and is rather due to a delay and/or abnormalities in nervous system development.     

The gametotoxic effect of antenatal exposure to thiotepa in mice]

ThioTEPA was injected intraperitoneally into female 101/H and CBA mice on day 12 of pregnancy at a dose of 2.5 mg/kg body weight. As a result, the number of prospermatogonia in 19-day-old male fetuses decreased to 78.8% in 101/H mice and to 63.7% in CBA mice. The gametotoxic effect on the male offspring (F1) in 101/H mice treated with thioTEPH included the significant decrease in the number of spermatogonia A, spermatocytes I at preleptotene and pachytene, as well as of spermatids at the stage 7 of development. Interstrain differences are described in the intensity of spermatogenesis in the mature intact animals and in regeneration capacity of gonads after prenatal thioTEPA administration.     

Spontaneous and induced chromosome aberrations in the bone marrow cells of different strains of mice during aging

Sensitivity of bone marrow cell chromosomes to alkylating agent thiophosphamide and to gamma-irradiation has been studied in the course of ageing in 101/H, A/He, CBA, BALB/c and C57BL/6 mice. The effects of both the kinds of mutagenic treatment and of the genotype of the animals on the age-dependent changes in sensitivity of bone marrow cell chromosomes were found. Following gamma-irradiation under our experimental conditions, no variation in the output of chromosomal aberrations was observed between the strains studied. Following thiophosphamide treatment, aged mice of strains 101/H, A/He and CBA showed an increased chromosome instability as compared to young ones. In C57BL/6 mice the level of induced chromosome aberrations was found to be age-independent. Following thiophosphamide treatment, cells with multiple chromosome lesions were found in the bone marrow. The higher instability of aged animals in some strains was mainly due to a sharp increase in the number of such cells. In the intact mice of all the strains studied no age-dependent increase in the number of cells showing structural chromosome aberrations was observed, while accumulation of aneuploid cells varied with genotype.     

A search for strain differences in response of mice to mutagenesis by thio-TEPA

After treatment of mice with thio-TEPA Malashenko and colleagues found differences among inbred strains in yield of dominant lethals and of chromosome aberrations in bone marrow, which they attributed to genes affecting repair. An attempt was made to confirm this work by comparing yields of dominant lethals in different strains of females mated to the same strain of males. However, no differences were found, all strain combinations giving 42-49% dominant lethals after a dose of 2 mg/kg thio-TEPA to late spermatids. Thus, the existence of genetic differences in repair of thio-TEPA induced lesions between strains CBA and C57BL/6J and between C3H/He and 101/H is not confirmed. Possible reasons for the discrepant results are discussed.     

Effect of antenatal action of Thio-Tepa on spermatogenesis of mature 101/N and CBA mice]

On pregnancy day 12 101/H and CBA mice were injected intraperitoneally 2.5 mg/kg bw thiophosphamide. 3.5-month-old male offspring were sacrificed. The drug effect on the testes was evaluated by karyologic analysis of the germ cell generations on stage 7 of the seminiferous epithelium cycle. A reliable reduction in the number of spermatogonia A, preleptotene and pachytene spermatocytes and spermatids at development stage 7 was found in 101/H mice. There were interspecific differences in spermatogenesis intensity in intact animals and recovery of germ cell pool after thiophosphamide action inducing toxicity.     

Strain-specific response in mice to the neonatal administration of ACTH(4-10) fragment: behavior, neurochemistry, and brain morphology

Neonatal injection of the ACTH4-10 fragment (5 micrograms daily for five days) caused genotype-dependent changes in concentrations of some monoaminergic neuromediators and their metabolites in hippocampus and brain stem of adult CBA and 101/HY mice. The catecholaminergic neurons increased in number in hypothalamic zona incerta of adult 101/HY mice. Neonatal injection of the peptide caused also genotype-dependent changes in the exploratory behavior of adult animals. Sound sensitivity was reduced in the 101/HY mice, whereas no sensitivity was revealed in both control and experimental groups of the CBA mice. The effects discovered were suggested to be caused by changes in neuronal differentiation.     

Structural abnormalities of hippocampus in 101/HY mice

We studied cytoarchitectonics of the hippocampus in 101/HY and CBA mice on brain sections stained after Nissl and Timm. In CBA mice, the structure of hippocampus was normal. In 10/HY mice, stratum pyramidale in field CA3 was "splitted" and the density of pyramidal neurons was decreased. Abnormalities were also found in the zone of suprapyramidal projections of mossy fibers (sp-ME), i.e., terminals of axons of the fascia dentata granular cells on the apical dendrites of pyramids. If in CBA mice the sp-MF zone was normal, i.e., looked like a vast compact formation or dense ordered bundle, in 101/HY mice, the sp-MF zone represented a group of scattered, diffuse, and interrupted bundles of varying length, some of which were incorporated in stratum pyramidale. Possible causes of the described morphological abnormalities are discussed, as well as their relation to specific features of biology, behavior, and neurological status of 101/HY mice.     

The activity of alkaline phosphatase in the sera of DBA/2NCrj and CBA/BrA mice with calcified tongue lesions

Spontaneously occurring calcified lesions were found in the tongues of DBA/2NCrj and CBA/BrA mice. In the DBA/2NCrj strain, the frequency of the lesion was 80% (males) and 88% (females). The youngest age of a mouse with this lesion was 18 days after birth, and 3-4 lesions were found in the tongue of 6- to 8-week-old mice. In CBA/BrA mice, 20% of females and 48% of males had the lesions. No significant differences were found in the serum calcium concentrations in high and low lesion-developing strains, but the alkaline phosphatase activities in the high-developing DBA/2NCrj, DBA/LiA, and CBA/BrA strains were higher than in strains with no calcified lesions.     

Relation of Infection to Tissue Temperature in Mice Infected with Mycobacterium marinum and Mycobacterium leprae

Intravenous and footpad infections with Mycobacterium marinum and footpad infections with M. leprae were compared in the following mouse strains: A/He, BALB/C, CBA, C3H, C57BL, C57L, DBA, 101, and CFW. The results varied a great deal according to mouse strain used. Intravenous injection of high doses of M. marinum resulted in deaths after 28 days of 100% of strain A/He, and none of strain 101; 27 days after injection, the feet and noses of all strain CBA mice, but few of the C57BL, 101, or CFW mice, were involved. Injection of a small dose of M. marinum into the footpad produced visible disease in 5 days in all of the C57BL and 101 mice, but in not more than 60% of the A/He, DBA, and CFW mice; the average amount of swelling at 17 days varied from 4.40 mm in strain C57L to 0.92 in strain 101. After footpad injection of M. leprae, the average plateau harvests varied from 1.3 x 10(7) acid-fast bacteria in strain CBA to 6.5 x 10(5) in strain C57L. The infections in CBA mice extended from the site of inoculation throughout the foot. The temperature was measured rectally, in the footpad, and in the tail. Analysis of all the results revealed little correlation among the three types of infection. There was a strong negative correlation between the tail temperature and the death rate after intravenous injection of M. marinum, and a strong positive correlation between footpad temperature and plateau harvest of M. leprae.     

Structural Abnormalities of Hippocampus in 101/HY Mice

We studied cytoarchitectonics of the hippocampus in 101/HY and CBA mice on brain sections stained after Nissl and Timm. In CBA mice, the structure of hippocampus was normal. In 101/HY mice, stratum pyramidale in field CA3 was splitted and the density of pyramidal neurons was decreased. Abnormalities were also found in the zone of suprapyramidal projections of mossy fibers (sp-MF), i.e., terminals of axons of the fascia dentata granular cells on the apical dendrites of pyramids. If in CBA mice the sp-MF zone was normal, i.e., looked like a vast compact formation or dense ordered bundle, in 101/HY mice, the sp-MF zone represented a group of scattered, diffuse, and interrupted bundles of varying length, some of which were incorporated in stratum pyramidale. Possible causes of the described morphological abnormalities are discussed, as well as their relation to specific features of biology, behavior, and neurological status of 101/HY mice.     

Changes in germ cell population in young and adult female mice from two inbred strains: CBA/Kw and KE

Female mice from two inbred strains CBA/Kw and KE differ markedly in fertility. The gametes of females from KE strain are of poorer quality than those of CBA/Kw. We analyzed the number of oocytes per ovary in KE and CBA/Kw mice aged 5, 25, 90, 180 and 360 days. The ovaries were dissected and processed according to the routine histological methods. In case of five-day-old females we used a modified distributed point counting method while in order to examine the gonads of older females, the nucleoli counting method was applied. In general, we observed gradual decrease in germ cell number throughout the whole life of females from both strains. The noticeable wave of oocyte loss occurs between 5th and 25th days of life. The mice from KE inbred strain on day 25th (1650 +/- 322 vs. 1140 +/- 210) and 90(th) (1040 +/- 211 vs. 692 +/- 89) days have significantly (p<0.005) more germ cells than the females from CBA/Kw strain. In older females the differences were not statistically significant. Interestingly, CBA/Kw females were found to have more rapid loss of primordial follicles throughout their lives. This can explain their shortened reproductive lifespan which was observed earlier.     

The remote effects of neonatal injections of caffeine and piracetam on audiogenic seizure susceptibility in mice of three genotypes

Neonatal DBA/2J, 101/HY and CBA/Lac/Sto mice (2-7-day-old) were subcutaneously injected with caffeine (200 mg/kg), piracetam (50 mg/kg) or distilled water. At the age of 1 month, they were tested for audiogenic seizure susceptibility (SS). The neonatal injections changed SS in 1-month-old mice in a genotype-dependent manner. Distilled water (control of neonatal pain stimulation) slightly reduced the audiogenic fit severity (arbitrary scores) the effect being most distinct in DBA/2J, less strong in 101/HY strain and absent in CBA. Caffeine neonatal injections induced slight changes in DBA/2J, no changes in CBA and increased SS in 101/HY mice. Piracetam reduced fit intensity in DBA/2J mice but increased it in CBA and, especially, in 101/HY strain. Genotype-dependent differences in physiological mechanisms of audiogenic seizures may be responsible for different remote effects of early treatment.     

Vasoformative non-osteogenic (angio) sarcomas of bone-marrow stroma due to strontium-90.

In a series of experiments, mainly CBA/H, but also C2H/H, mice aged 3 months were injected intraperitoneally with solutions of 90Sr Cl2, the dose per mouse varying from 7 to 20 muCi, and compared with similar mice treated with 226Ra or 239Pu, discussed elsewhere. In male mice, the commonest tumour resulting at each dose of 90Sr was non-osteogenic (angio) sarcoma, a tumour not seen after 226Ra. In females, this tumour occurred far less frequently than osteosarcoma. In CBA mice of both sexes converted to radiation chimaeras (which are sterile) and similarly treated with 90Sr, the only skeletal tumours were osteosarcomas. When only half the body of CBA mice was X-irradiated with 1000 rad and the mice given 90Sr, non-osteogenic sarcoma occurred predominantly in those mice X-irradiated in the cephalic half. The results suggest that intact testes may provide co-factors for this type of neoplasm, whereas others have shown that oestrogens facilitate murine osteosarcoma. The non-osteogenic osteosarcomas arise from damaged stromal elements in bone-marrow of selected bones. The risk to this component of bone-marrow, as well as to haematopoietic tissue, should be considered in radiation protection.     

The effect of prodigiozan on the postradiational recovery of hemopoiesis in long-term bone morrow cultures of mice of different genotypes

The influence of prodigiozan on the processes of postirradiation recovery of hemopoiesis in long-term bone morrow cultures of two strain mice, having genetic distinctions in the condition of systems of the reparation DNA was investigated. It was showh, that the irradiation of long-term bone morrow cultures of mice reparation-defective strain 101/H resulted in the greater degree damage of early haemopoietic precursors (GM-CFC), reduction of the amount of the immature and of the mature granulocytes and of the decrease of the number of stromall cells in the comparison with the bone morrow of reparation-capable mice (CBA x C57B1)F1. Under the introduction in cultures of prodigiozan for 24 hours prior to an radiation the distinctions of the speed of postirradiation recovery of hemopoiesis substantially smoothed out, and the protective effect of the drag in bone morrow cultures of mice 101/H was comparable to those, marked in bone morrow cultures of reparation-capable strain mice (CBA x C57B1)F1. It is supposed, that this effect can be caused by the activation of the hematopoietic microenvironment cellular elements and inclusion of the mechanisms of intercellular of interactions, which provide stimulation of the regenerative processes at action radioprotective drags and can in the certain degree to compensate the defect of the systems of the reparation DNA.     

Lethality in LPS-induced endotoxemia in C3H/HeCr mice is associated with prevalence of proinflammatory cytokines: lack of protective action of lactoferrin

C3H/HeCr mice are more susceptible to infection compared with other strains. Lactoferrin (LF), a protein involved in innate defense, was shown to protect mice against lethal endotoxemia. In this investigation we attempt to explain the cause of increased susceptibility of C3H/HeCr mice to LPS and lack of protective LF action in these mice. We found that C3H/HeCr mice produced up to 5-fold more serum TNFalpha and 66% higher IFNgamma levels in response to i.v. LPS injection than the control, CBA strain. 24 h pretreatment of C3H/HeCr mice with LF did not cause inhibition of the LPS-induced TNFalpha serum levels, whereas in CBA mice LF significantly decreased TNFalpha level. IL-6 serum levels, in turn, were lowered in C3H/HeCr mice but elevated in CBA mice. That differential regulation of cytokine production by LF in C3H/HeCr mice paralleled a decreased survival after lethal LPS injection - 10% vs. 60% in control, PBS treated mice. In addition, determination of colony forming units (CFU) in livers and spleens after administration of 10(8) Escherichia coli revealed that pretreatment of CBA mice with LF caused a marked reduction of CFU in these organs, whereas in C3H/HeCr mice the changes were insignificant. These results indicate that the altered TNFalpha/IL-6 ratio in C3H/HeCr mice, as compared to control CBA mice, as well as the increased IFNgamma level, may be responsible for the increased susceptibility to endotoxemia in that substrain. We also suggest that an association exists between the LF protective effect against endotoxic sequelae and the insult-induced systemic immune response.     

A new alloantigenic system associated with the Mls locus in the mouse.

An antiserum prepared by injecting C3H/HeJ mice with CBA/J tissue has been shown to react with cell surface components that are not part of any previously described system of serologically detectable alloantigens. The antiserum, which is designated AST-101, acts selectively in cytotoxic tests carried out with lymphoid cells, killing B cells, but not T cells. Phagocytic cells found in peritoneal exudates are also killed by AST-101 and complement in vitro; the sensitivity of other cell types has not been determined. Strain distribution does not indicate any association of the AST-101 system with H-2, Ly, or Thy systems; genetic analysis reveals close linkage with the mouse minor MLC-stimulating (Mls) locus. Serologic analysis also points to a close association between antigens reactive with AST-101 and the products of the Mls genes.     

A potent modifier of liver cancer risk on distal mouse chromosome 1: linkage analysis and characterization of congenic lines

The C3H/HeJ (C3H) and CBA/J (CBA) mouse strains are classical mouse models of cancer susceptibility, exhibiting high risks for both spontaneous and chemically induced liver cancer. By analysis of backcrosses and intercrosses between C3H or CBA and resistant B6 mice, we have mapped a potent modifier of hepatocellular carcinoma development to distal chromosome 1, linked to the marker D1Mit33 with combined LOD(W) scores of approximately 5.9 (C3H) and 6.5 (CBA). We previously identified this region as one of two that modify susceptibility in the more distantly related C57BR/cdJ (BR) strain. Congenic B6.C3H(D1Mit5-D1Mit17) and B6.BR(D1Mit5-D1Mit17) mice developed significantly more liver tumors than B6 mice did (6- to 13-fold, P < 10(-11), in males; 3- to 4-fold, P < 10(-3), in females). Thus, distal chromosome 1 carries one or more genes that are sufficient to confer susceptibility to liver cancer.     

Preimplantation embryonic mortality--method of studying interstrain differences in the reparative activity of mouse ova

Cytological analysis of preimplantation embryonic death in 101/HY, C57BL/6JY and CBA/lacY females crossed with hybird males F1 (BALB/cYxDBA/2Y) was carried out. Embryonic death was induced by thiophosphamide at a dose of 2 mg/kg, i. p. The maximum preimplantation death was recorded in 101/HY females (38.8%), the minimum in CBA/LacY females (21.9%). In C57BL/6JY females, the maximum preimplantation death accounted for 31.3%. Thus the same chromosome damage induced by thiophosphamide in late spermatids of F1CD2 males caused quantitative differences in embryonic mortality in females of different genotypes. The data obtained evidence that fertilized eggs are capable of repairing part of damage induced by paternal chromosomes. It was demonstrated that the preimplantation embryonic death can be used for studying strain differences from the reparative activity of mouse eggs.     

Marked difference in sensitivity to gamma-ray-induced cataract between BALB/c and CBA-C3H strain mice]

Nineteen BALB/c, 14CBA/KI, 12C3H/HeJ and 15 (CBA/Kl x C3H/HeJ) F1 female mice were irradiated with 850 rad gamma-rays and transferred with 10(7) syngeneic bone marrow cells 24 hrs later. The occurrence of cataract was examined in these animals. All the BALB/c mice showed visible lens opacification in both eyes between 113 and 149 days after irradiation. All the animals were autopsied 6 months after irradiation and examined for opacification of their lenses. The proportion of opaque lenses was 100, 7.1, 16.7 and 0% in BALB/c, CBA/Kl, C3H/HeJ and (CBA/Kl x C3H/HeJ) F1 mice, respectively. The results indicate that BALB/c mice are much more sensitive to radiation-induced cataractogenesis than CBA and C3H mice.     

Molecular manifestations of radiation-induced genome instability: the possibility of chemical modification

The molecular manifestations of radiation-induced genome instability-changes of the DNA structure, the excision DNA repair and the contents of the reactive oxygen forms in bone marrow cells of the repair proficient mice (CBA) and of the repair-defective (101/H) lines in the dynamics up to 185 day after ionizing radiation exposure in the dose of 1.5 Gy were studied. Is was established, that after irradiation in bone marrow cells the descendants with the decreased activity of excision DNA repair and prone to increased changes of DNA structure DHK is arised. The injection of the phenozane in concentrations causing its receptor interaction with cells, did not defend DNA of the bone marrow cells from the radiation injury after the exposure in a sublethal dose, however it exerted influence on long-term changes. Due to the phenosane of the bone marrow cells of the irradiated mice of CBA line exhibited the larger activity in a DNA repair from damages and maintenance of vitality. The bone marrow cells of male mice of repair defective 101/H line, which phenozan was entered before the irradiation, remained unfit to the remuval of DNA damages by the repair, that probably resulted the activations of the program of the maintenance of genome constancy by the apoptosis in the cells--carriers of the structural defects and the cause of animal lethality.