Comparative serum metabolomics between SCID mice and BALB/c mice with or without Schistosoma japonicum infection: Clues to the abnormal growth and development of schistosome in SCID mice.

The small blood flukes of genus Schistosoma, which cause one of the most prevalent and serious parasitic zoonosis schistosomiasis, are dependent on immune-related factors of their mammalian host to facilitate their growth and development, and the formation of granulomatous pathology caused by eggs deposited in host's liver and intestinal wall. Schistosome development is hampered in the mice lacking just T cells, and is even more heavily retarded in the severe combined immunodeficient (SCID) mice lacking both T and B lymphocytes. Nevertheless, it's still not clear about the underlying regulatory molecular mechanisms of schistosome growth and development by host's immune system. This study, therefore, detected and compared the serum metabolic profiles between the immunodeficient mice and immunocompetent mice (SCID mice vs. BALB/c mice) before and after S. japonicum infection (on the thirty-fifth day post infection using liquid chromatography-mass spectrometry (LC-MS). Totally, 705 ion features in electrospray ionization in positive-ion mode (ESI+) and 242 ion features in ESI- mode were identified, respectively. First, distinct serum metabolic profiles were identified between SCID mice and BALB/c mice without S. japonicum worms infection. Second, uniquely perturbed serum metabolites and their enriched pathways were also obtained between SCID mice and BALB/c mice after S. japonicum infection, which included differential metabolites due to both species differences and differential responses to S. japonicum infection. The metabolic pathways analysis revealed that arachidonic acid metabolism, biosynthesis of unsaturated fatty acids, linoleic acid metabolism, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, alpha-linolenic acid metabolism, glycerophospholipid metabolism, sphingolipid metabolism and purine metabolism were enriched based on the differential serum metabolites between SCID mice and BALB/c mice after S. japonicum infection, which was addressed to be related to the retarded growth and development of S. japonicum in SCID mice. These findings provide new clues to the underlying molecular events of host's systemic metabolic changes on the growth and development of S. japonicum worms, and also provide quite promising candidates for exploitation of drugs or vaccines against schistosome and schistosomiasis.

Strain- and sex-linked effects of dietary polyunsaturated fatty acids on tumor growth and immune functions in mice.

In the present paper, we studied the influence of different levels of dietary polyunsaturated fatty acids (PUFA) on the immune system and tumor growth in young mice. Female BALB/c mice fed a PUFA-rich diet display an enhanced body growth, proliferative response to mitogens in vitro, and rate of growth of a spontaneous transplantable adenocarcinoma as compared to PUFA-poor diet-fed females. Such effects are, however, limited by sex and strain background genes located outside the H-2 complex. In effect, the influence of dietary PUFA content is evident in female but not in male BALB/c mice. Moreover, in female DBA/2 mice with the same haplotype (H-2d) of the major histocompatibility complex as that of BALB/c mice, low dietary PUFA determines a reduced tumor growth only, but it does not affect body growth and proliferative response to mitogens in vitro.

Expression of specific glycoconjugates in both primary and secondary olfactory pathways in BALB/C mice.

Binding of cell surface carbohydrates to their receptors specifically promotes axon growth and synaptogenesis in select regions of the developing nervous system. In some cases these interactions depend upon cell-cell adhesion mediated by the same glycoconjugates present on the surface of apposing cells or their processes. We have previously shown that the plant lectin Dolichos biflorus agglutinin (DBA) binds to a subpopulation of mouse primary olfactory neurons whose axons selectively fasciculate prior to terminating in the olfactory bulb. In the present study, we investigated whether these glycoconjugates were also expressed by postsynaptic olfactory neurons specifically within the olfactory pathway. We show here for the first time that DBA ligands were expressed both by a subset of primary olfactory neurons as well as by the postsynaptic mitral/tufted cells in BALB/C mice. These glycoconjugates were first detected on mitral/tufted cell axons during the early postnatal period, at a time when there is considerable synaptogenesis and synaptic remodelling in the primary olfactory cortex. This is one of the few examples of the selective expression of molecules in contiguous axon tracts in the mammalian nervous system. These results suggest that glycoconjugates recognized by DBA may have a specific role in the formation and maintenance of neural connections within a select functional pathway in the brain.

Studies of sequence heterogeneity of mitochondrial DNA from rat and mouse tissues: evidence for an increased frequency of deletions/additions with aging.

To obtain information on the extent of random nucleotide changes in mitochondrial DNA (mtDNA) from different organs of young adult and senescent Fischer 344 rats, the temperature of thermal denaturation (tm) was measured in (1) the native mtDNA cut at a single SstI site and (2) the reannealed duplexes formed after the initial melting of the mtDNA sample. No change was found between the two tm values in either young or senescent mtDNA, suggesting that the overall mismatch in nucleotide sequence in these samples was below the resolution of the method estimated at about 0.2%. In another experiment, mtDNA samples from young adult or senescent BALB/c mouse liver were digested with EcoRI, denatured and allowed to reanneal. The duplexes formed by the 14-kb EcoRI fragment were analyzed in randomly taken electron micrographs for the occurrence of mismatched segments. About 1.8% of reconstituted duplexes in adult mtDNA and 11% of those in senescent mtDNA contained small loops or knobs suggestive of deletions/additions of about 400 +/- 150 nucleotides. These data correspond to about 1% of the native mtDNA population in adult liver and about 5% in senescent liver having deleted/inserted segments. Although deletions/insertions may occur at variable sites, their distribution appears to be non-random. These findings suggest that small sequence rearrangements, which have been observed previously in unicircular dimers of mouse and human mtDNA, occur also in monomeric mtDNA from normal tissues and accumulate with aging.

Marked differences in olfactory sensitivity and apparent speed of forebrain neuroblast migration in three inbred strains of mice.

In the adult forebrain, new neuroblasts constantly migrate from the subventricular zone along the rostral migratory stream to the olfactory bulb, where many become neurons. It is unclear whether this process is different in commonly used mouse strains and whether it is related to olfactory function. Adult male BALB/c, C57BL/6, and 129/S1 (formerly 129SV) mice were tested for olfactory sensitivity plus discrimination, using male mouse urine from the two other strains. BALB/c mice had the greatest olfactory sensitivity, followed by 129/S1, and C57BL/6 mice, by an order of magnitude each. Newly formed cells were pulse-labeled for 3 h with i.p. 5-bromo-2'-deoxyuridine (BrdU) injections and the animals analyzed 24 h later. In 129/S1 mice, a greater proportion of neuroblasts were present closer to the olfactory bulb than in BALB/c mice, followed by C57BL/6 mice. The total number of BrdU-labeled cells did not differ, suggesting differences in migration and not proliferation. The impaired olfactory function in C57BL/6 mice might be caused by the reduced number of neuroblasts that reach the olfactory bulbs. However, olfactory function in BALB/c and 129/S1 mice did not correlate with their putative migration speed, suggesting a more complex nature of cellular processes that contribute to olfactory function. These results caution against comparing studies of olfactory function or neural precursors that use different strains of mice, and question the use of C57BL/6 mice as a "normal" strain or as transgenic background. Perhaps more importantly, the results point to an opportunity to identify genes that regulate olfactory function and neuroblast behavior.

Expression and distribution of the receptors for coxsackievirus B3 during fetal development of the Balb/c mouse and of their brain cells in culture.

This study was designed mainly to determine the relationships between the expression and distribution of the cellular receptor proteins for coxsackievirus B3 (CVB3) and susceptibility of mouse brain cells during fetal development of Balb/c mice. Immunoblot analysis of fetal extracts demonstrated that the CVB3 receptor proteins were first expressed at day 14 of the fetal stage, and that maximal expression of the cellular receptor occurred at near term or newborn stage. Results also suggested that newborn mouse brain tissue expressed much larger quantities of viral receptor proteins, compared to other tissues. In vitro studies showed that both mouse neurons and astrocytes could be infected by two CVB3 strains, pantropic CVB3 Nancy strain (CVB3N) and myocardiotropic CVB3 Woodruff strain (CVB3W). CVB3N, however, replicated and grew to high titer in primary astrocyte cultures and in primary neuron cultures, whereas, primary astrocyte cultures were relatively resistant to CVB3W. Virus binding assays revealed that CVB3N bound faster and in greater amounts to mouse brain cells than CVBW. These two virus strains, however, were found to share the same receptor specificity by virus competition assays. The number of virus binding sites for CVB3 on newborn mouse brain cells was approximately 1.8 x 10(4) per cell. The data suggested that preferential expression of the cellular receptors on newborn mouse brain cells may be related to their high susceptibilities to CVB3 infection.

Age-related suppression of murine lymphoid cell blastogenesis by marijuana components.

Marijuana components modulate a variety of immune response parameters. The cannabinoids delta 9-tetrahydrocannabinol (THC) and 11-hydroxy-tetrahydrocannabinol (11 OH-THC) are known to depress the in vitro proliferative response of murine lymphoid cells to the mitogens concanavalin A (Con A) and phytohemagglutinin (PHA). In the present report the effects of THC and 11 OH-THC on adult thymus and spleen cells were compared to effects on lymphoid cells of those organs from juvenile mice at various ages. The results demonstrate differences in susceptibility to cannabinoid-induced suppression by lymphoid cells from different organs and different age mice. In adults, thymus cells were suppressed more readily than spleen cells. Splenocytes from mice under 2 weeks old were suppressed much more readily than those from older mice. Cell populations from organs with higher proportions of L3T4+/Lyt2- cells were more difficult to suppress. The possible mechanisms involved and directions for future work are discussed.

The alpha 1, alpha 2, and alpha 3 subunits of GABAA receptors: comparison in seizure-prone and -resistant mice and during development.

Gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in brain, opens chloride channels through actions on GABAA receptors. We now report base and amino acid sequences of the alpha 1, alpha 2, and alpha 3 subunits from GABAA receptors of audiogenic seizure-prone (DBA/2J) and -resistant (C57BL/6J) inbred strains of mice. Inbreeding had fixed different alleles of the alpha 1 subunit in the two strains, giving five base differences in the cDNAs. None of these affected amino acid sequence, but one did create a NsiI restriction site potentially useful in mapping genomic DNA. No base or amino acid sequence differences between the strains were detected for the other two subunits. Northern blots revealed no apparent strain differences in message levels for these three subunits in whole brains of the mice at 3 weeks of age, the peak of seizure susceptibility in DBA/2J, but did reveal distinct regional and developmental patterns of expression among the subunits in mouse brain.

Strain-dependent decrease in glutamate binding to the N-methyl-D-aspartic acid receptor during aging.

Glutamate binding to the N-methyl-D-aspartic acid (NMDA) receptor decreases in two strains of aged mice. In BALB/c mice the Bmax values decline 16% by 10 months and 45% by 30 months of age when compared to 3 months. The Kd increased more by 10 months (+29%) than by 30 months (+14%). In the C57Bl strain the Bmax was unaltered by 10 months but decreased 17% by 30 months. The Kd values, however, increased 121% by 10 months and 283% by 30 months of age. These data suggest that the age-related decline in glutamate binding to the NMDA receptor may predict a strain-specific reduction in NMDA-mediated processes (e.g. long-term potentiation, postsynaptic calcium fluxes).

Age and strain dependence of O6-methylguanine DNA methyltransferase activity in mice.

The activity of the DNA repair protein O6-methylguanine DNA methyltransferase (MT) was compared in liver extracts from female ICR and male C57BL/6 mice at various ages (3-130 weeks old). Similar patterns of overall enzyme activity were observed in both strains with O6-MT activity being relatively low in young mice (3 or 8 weeks old). However, the activity significantly increased after adolescence (middle age), thereafter decreasing with old age (over 100 weeks old) to a level equivalent to that found in young mice. In an additional strain difference study, O6-MT activities in liver extracts from 4 strains of mice were compared at 5 and 30 weeks of age. Although a similar age-associated increase of enzyme activity in adolescence was confirmed in all 4 strains investigated, the closed-colony ICR mice differed from the inbred strains in demonstrating significantly higher levels of O6-MT activity in females than in males. However, the same tendency was also observed in a comparison of the sexes in 30-week-old C3H/HeN, C57BL/6 and BALB/c mice.

Population density and growth rate in laboratory mice.

Home Office guidelines recommend an area of 60 cm2 per mouse for growing mice up to 30 g. However, the overall growth rate, and final adrenal weight of weanling BALB/c and MF1 strain mice was not affected by being housed at a density of down to 27 cm2 per mouse, though there was some evidence of strain differences in ability to tolerate such dense housing. The presence of cage accessories had no effect on growth rate of BALB/c and female mice, but reduced growth of MF1 and male mice, though the effect was small. It is concluded that present Home Office guidelines make a generous provision for the space requirements of growing laboratory mice, and that the use of cage accessories of varying design may be worth exploring in more detail.

Aspen wood-wool is preferred as a resting place, but does not affect intracage fighting of male BALB/c and C57BL/6J mice.

Aspen wood-wool, provided as nesting material, was evaluated as a possible improvement of cage environment for 10-14-week-old inbred male mice maintained in groups of six (BALB/c n = 72 and C57BL/6J n = 36). The daily behaviour of mice was video recorded and their body weight, food consumption, weights of some organs and serum corticosterone concentrations were measured. Aggressive interactions between cage mates and against a strange intruder as well as the number of wounds on the back of the animals was monitored in order to evaluate the effect of nesting material on intermale aggression. Nesting material did not affect the daily active/passive behaviour patterns of mice, although animals clearly preferred it as a resting place. BALB/c mice given nesting material showed less weight gain and smaller brown adipose tissue weights than animals without nesting material. The other characteristics measured were not affected by the presence of nesting material in either strain. The presence of nesting material had no effect on fighting in cages. C57BL/6J mice were more aggressive than BALB/c mice according to the number of wounded animals in a cage. Wounded BALB/c mice had enlarged spleens and decreased epididymal adipose tissue weights. In conclusion, the nesting material used in this study did not adversely affect the animals. On the other hand, the material was clearly preferred to conventional bedding as a resting place. These findings suggest that nesting material may improve the cage environment of laboratory mice. Furthermore, there was an indication of strain differences in aggressive behaviour. It could be suggested that C57BL/6J mice are less tolerant towards intruders and housing six mice per cage is not suitable for this strain.

[Interstrain differences in the serotonin and 5-hydroxyindoleacetic acid levels in the postnatal ontogeny of C57BL/6J and BALB/cLac strain mice]. / Mezhlineinye razlichiia v urovne serotonina i 5-gidorksiindoluksusnoi kisloty v postnatal'nom ontogeneze myshei linii C57BL/6J i BALB/cLac.

The content of serotonin and 5-hydroxy-indole-acetic acid (5-HIAA) was determined in the brain stem and hemispheres in 1 and 3 months old C57BL/6J and BALB/cLac mice. The characteristic dynamics of serotonin and its metabolite content related to the age was found in different brain regions and proved to be similar in both the strains, but the rate of serotonin system development in C57BL/6J mice was higher than in BALB/c Lac mice. An intensive catabolism of serotonin, possibly, related to the reaction to new environment was noted in the newborn animals. Sex differences in the rate of serotonin system maturation and serotonin and 5-HIAA content were shown for 12--16 days old mice: 12 days old males were characterized by more intensive metabolism than females while 16 days old males had less serotonin than females.

Sociability and brain development in BALB/cJ and C57BL/6J mice.

Sociability--the tendency to seek social interaction--propels the development of social cognition and social skills, but is disrupted in autism spectrum disorders (ASD). BALB/cJ and C57BL/6J inbred mouse strains are useful models of low and high levels of juvenile sociability, respectively, but the neurobiological and developmental factors that account for the strains' contrasting sociability levels are largely unknown. We hypothesized that BALB/cJ mice would show increasing sociability with age but that C57BL/6J mice would show high sociability throughout development. We also hypothesized that littermates would resemble one another in sociability more than non-littermates. Finally, we hypothesized that low sociability would be associated with low corpus callosum size and increased brain size in BALB/cJ mice. Separate cohorts of C57BL/6J and BALB/cJ mice were tested for sociability at 19-, 23-, 31-, 42-, or 70-days-of-age, and brain weights and mid-sagittal corpus callosum area were measured. BALB/cJ sociability increased with age, and a strain by age interaction in sociability between 31 and 42 days of age suggested strong effects of puberty on sociability development. Sociability scores clustered according to litter membership in both strains, and perinatal litter size and sex ratio were identified as factors that contributed to this clustering in C57BL/6J, but not BALB/cJ, litters. There was no association between corpus callosum size and sociability, but smaller brains were associated with lower sociability in BALB/cJ mice. The associations reported here will provide directions for future mechanistic studies of sociability development.

Increased juvenile and adult body weights in BALB/cByJ mice reared in a communal nest.

Both wild and laboratory mice and rats preferentially rear their young in communal nests and indiscriminately nurse any of the young within the nest. In this study, BALBc/ByJ mice reared under communal nesting (CN) conditions (3 dams and their litters sharing a common nest) were compared with BALBc/ByJ mice raised in single (one dam with her litter) nests (SN) in body weight from birth into adulthood; food and water intake and body composition were compared between adult mice. Compared with SN female mice, female CN mice (measured only until weaning) exhibited significantly higher body weights at postnatal days 11 and 25. Male CN mice were significantly heavier than were male SN mice at postnatal day 25 and at 20, 26, and 30 wk of age. There were no differences between adult male mice from CN and SN groups in 48-h food and water intake or body composition (total lean:total fat ratio; measured by quantitative MRI). In conclusion, BALB/cByJ mice reared under communal nesting conditions showed more robust juvenile growth rates than did mice raised with a single dam and litter per cage. In addition, body weights of male CN mice remained higher than male SN mice into adulthood.

Paternal transmission of complex phenotypes in inbred mice.

BACKGROUND: Inbred mice are genetically identical but nonetheless demonstrate substantial variability in complex behaviors such as activity levels in a novel environment. This variability has been associated with levels of parental care experienced early in development. Although maternal effects have been reported in biparental and uniparental strains, there have been no investigations of paternal effects in non-biparental strains in which offspring are reared exclusively by mothers. METHODS: In the uniparental inbred Balb/cJ mouse strain, we examined the relationship of paternal open-field activity to the activity of both male and female offspring in the open-field. Potential mediators of paternal transmission of behavior were examined, including maternal care, growth parameters, litter characteristics, and time the father was present with the pregnant mother prenatally. RESULTS: An association of paternal open-field activity with the open-field activity of female but not male offspring was found. Variation in maternal postnatal care was associated with female but not male offspring activity in the open-field but did not mediate paternal effects on offspring behavior. Paternal effects on offspring growth parameters were present, but these effects also did not mediate paternal effects on behavior. CONCLUSIONS: Paternal transmission of complex traits in genetically identical mice reared only by mothers suggests a nongenetic mechanism of inheritance potentially mediated by epigenetic factors. The exclusion of multiple mediators of paternal effects on offspring suggests the possibility of germline paternal inheritance via sperm of complex phenotypes in inbred mice. Future studies are required to examine these interesting possibilities.

Teratogenic effect of lithium carbonate in early development of BALB/c mouse.

Lithium carbonate is used as a standard treatment for manic depression. While researchers have investigated the teratogenic effects of lithium carbonate on embryos of various animals in later stages of development, very limited work has been done on the probability of effects at early stages of development. In this study, the teratogenic effect of lithium carbonate was investigated at earlier preimplantation through implantation stages of development of Balb/C mouse embryos. A therapeutic dose (i.e., 300 mg/kg b.w.) was injected into mice intraperitoneally on days 3.5, 4.5, 5.5, and 6.5 of pregnancy. Then, on day 15.5 of gestation, embryos were collected from the pregnant animals. Among the embryos, 71.7% were healthy, 10.7% resorbed, 3.1% showed lordosis, 8.1% were underdeveloped and 8.4% had eye malformations. Significant increases (P < 0.05) in the number of hepatic megakaryocytes and nucleated red cells were also observed among experimental embryos. Analysis of maternal serum proteins prepared from dissected animals showed a significant increase or decrease (P < 0.05) in the levels of serum proteins albumin, alpha2 globulin, beta globulin, and gamma globulin. This research on early developmental stages suggests that pregnant mothers need to be aware of possible teratogenic effects at early stages of pregnancy, although it has been thought that the egg envelope can prevent teratogens from entering. In this case, mothers may need to stop lithium carbonate treatment before they make a decision to become pregnant.

Study of embryotoxicity of mentha piperita L. during organogenesis in balb/c mice / Estudio de la embriotoxicidad de mentha piperita L. durante la organogénesis en ratones balb/c

Mentha piperita (Labiatae), commonly known as peppermint is a native Iranian herb which is used in folk medicine for various purposes. This study was carried out to reveal the teratogenic effect of Mentha piperita on mice fetuses. In this experimental study, pregnant Balb/c mice divided to four groups. Case group received 600 (treatment I) and 1200 (treatment II) mg/kg/day the hydroalcoholic extract of Mentha piperita during 6-15 of gestational days and one control group received normal saline during GD6-GD15 by gavages and other control group did not receive any matter during 6-15 of gestational days. Mice sacrificed at GD18 and embryos were collected. Macroscopic observation was done by stereomicroscope. 20 fetuses of each group were stained by Alizarin red-S and Alcian blue staining method. The Mean weight of fetuses decreased in treatment groups rather than control (P<0.05) but CRL there was no significant difference between treatments and controls groups. In the treatment I (600 mg/kg/day) and treatment II (1200 mg/kg/day), normal saline and control group, no gross congenital malformations were observed in fetuses. Treated fetuses also had no delayed bone ossification as determined by Alizarin red-S and Alcian blue staining method. This study showed that the hydroalcoholic extract of Mentha piperita (600 and 1200 mg/kg/day) has no teratogenic effect in mice fetuses if used continuously during embryonic period.

Mentha piperita (Labiatae), comúnmente conocida como menta, es una hierba nativa de Irán, que se utiliza en la medicina tradicional para diversos fines. Este estudio fue realizado para descubrir el efecto teratogénico de la Mentha piperita en fetos de ratones. Los ratones Balb/c preñadas fueron divididas en cuatro grupos. Los grupos recibieron 600 (tratamiento I) y 1200 (tratamiento II) mg/kg/día del extracto hidroalcohólico de Mentha piperita durante los días 6-15 de gestación (DG), mientras que un grupo control recibió solución salina normal durante los DG 6-15 vía oral y otro grupo control sano no recibió substancia durante los DG 6-15. Los ratones fueron sacrificados el DG 18, recolectando los fetos. Se realizó la observación macroscópica mediante un estereomicroscopio. 20 fetos de cada grupo se tiñeron por el método de rojo de alizarina-S y azul de Alcián. La media de peso de los fetos disminuyó más en los grupos de tratamientos que los controles (p <0,05), pero CRL no presentó diferencias significativas entre los tratamientos y los grupos control. En los fetos del grupos tratamiento I (600 mg/kg/día), tratamiento II (1200 mg/kg/día), solución salina normal y control no se observó ninguna malformación congénita grave. Los fetos tratados tampoco tuvieron osificación ósea retrasada según lo determinado por el método de rojo de alizarina-S y azul de Alcián. Este estudio mostró que el extracto hidroalcohólico de Mentha piperita (600 y 1200 mg/kg/día) no tiene efectos teratogénicos en fetos de ratones al ser utilizado continuamente durante el período embrionario.

The effects of neonatal vestibular stimulation on adult growth and emotional reactivity in BALB/c mice.

Two experiments were conducted to determine the effects of neonatal vestibular stimulation on the subsequent development and behavior in BALB/c mice. No evidence appeared to suggest that neonatal vestibular stimulation alters either the growth pattern or the ontogenesis of emotional reactivity.