Cerebral, cerebellar, and brain stem gangliosides in mice susceptible to audiogenic seizures

Abstract— The quantitative and qualitative distribution of gangliosides was investigated in the cerebrum, cerebellum and brain stem of audiogenic seizure resistant (C57BL/6J) and susceptible (DBA/2J) mice at 21 days of age. The concentration of gangliosides (μg/unit weight) was higher in the DBA cerebrum and brain stem, but lower in the DBA cerebellum compared to the concentration in C57 mice. In general, the brain water content was lower in DBA mice than in C57 mice. The distributions of a number of gangliosides were found to be different between the two strains and the differences were often in the same direction across the three brain regions. The most consistant and significant difference in ganglioside pattern observed between the strains was the higher concentration of G_M1 in all three regions of the DBA brain. These results suggest that DBA mice have a more heavily myelinated CNS than C57 mice. The relationship of these observations to inherent audiogenic seizure susceptibility is discussed.     

Ontogeny of susceptibility to the convulsant, Ro 5-4864, and its relationship to audiogenic seizure susceptibility in inbred mice

The postnatal development of susceptibility to the convulsant effects of Ro5-4864 (4'-chlorodiazepam) was characterized in two inbred mouse strains (DBA/2J and BALB/c ByJ) which as adults differ markedly in their response to this convulsant. Onset of susceptibility to a dose of Ro5-4864 which caused a high frequency of clonic seizures in adults was observed at 10 days of age in DBA/2 mice, but not until 35 days in BALB/c By mice. At 14 days of age an abrupt increase in susceptibility to Ro5-4864-induced tonic seizures was found in DBA/2 but not BALB/c By mice. Both the peak of tonic seizure susceptibility (21 days) and the time course of its subsequent age-dependent decline closely paralleled the change in audiogenic seizure susceptibility in the DBA/2 strain. PK11195 (40 mg/kg) blocked Ro5-4864 (25 mg/kg)-induced, age-dependent tonic seizures but had no effect on clonic seizure induction in the same mice. These observations establish that both the susceptibility to Ro5-4864 in adult mice and the postnatal time course for development of susceptibility to this convulsant are inherently different in these two strains of mice. The lack of coincidence between the developmental onset of susceptibility to Ro5-4864-induced seizures and the onset of supersensitivity to Ro5-4864-induced tonic seizures during the period of peak audiogenic seizure susceptibility in DBA/2 mice implies that more than one neurochemical mechanism is involved in the ability of Ro5-4864 to induce seizures in this strain. However, the blockade of Ro5-4864-induced tonic seizures by PK11195 suggests that peripheral type benzodiazepine receptors may mediate this effect.     

Pregnancy rates, prenatal and postnatal survival of offspring, and litter sizes after reciprocal embryo transfer in DBA/2JHd, C3H/HeNCrl and NMRI mice

Success of embryo transfer is often a limiting factor in transgenic procedures and rederivation efforts, and depends on the genetic background of the donor and recipient strains used. Here we show that embryo transfer to DBA/2J females is possible, and present data on pre- and postnatal success rates after reciprocal embryo transfer using the inbred DBA/2J and C3H/HeN, and outbred NMRI strains. The highest embryo yield was achieved in outbred NMRI females, but embryo yields were similar in DBA/2J and C3H/HeN mice following superovulation despite poor estrus cycle synchronization in DBA/2J females. In-strain transfer of DBA/2J blastocysts (transfer of embryos to recipients from the same strain) resulted in pregnancy rates (57.1%) similar to those obtained following in-strain transfer of C3H/HeN (60.0%) and NMRI mice (83.3%), although the prenatal survival rate of blastocysts was low. Moreover, from the pups born only half survived the postnatal period after transfer of DBA/2J and C3H/HeN blastocysts to DBA/2J recipients. These problems were not observed when transferring NMRI-blastocysts to C3H/HeN and DBA/2J mothers. The number of blastocysts transferred also had a positive effect on the success of embryo transfer. In conclusion, C3H/HeN and DBA/2J females can be used as recipients for embryo transfer procedures for certain donor strains like NMRI, as one major determinant seems to be the genetic background of the embryos transferred. We also recommend to increase the number of DBA/2J blastocysts transferred, and to foster the DBA/2J pups to other DBA/2J mothers postnatally for in-strain transfer of DBA/2J mice.     

Genetic Differences in Androgen Receptors and in Autoregulation of Testicular Human Chorionic Gonadotropin Binding Sites in the Mouse

Abstract

The autoregulation of testicular human chorionic gonadotropin (hCG) binding sites was studied in two strains of mice known to differ in their endocrine and reproductive characteristics (C57BL/10J and DBA/2J), and in their F₁ progeny (B10D2F₁). Basal hCG binding levels were higher in C57BL/10J than in DBA/2J mice, while B10D2F₁ mice had intermediate levels. Twenty-four h after injection, hCG produced dose-related changes in hCG binding in C57BL/10J and B10D2F₁ mice not observed in DBA/2J mice. However, 72 h after treatment with hCG there was a decrease in hCG binding in all the strains studied. These results suggest the participation of genetic factors in determining basal levels, dose-related changes and temporal response of testicular hCG binding sites to hCG administration. Androgen receptor levels were measured in the same strains of mice. DBA/2J mice had higher receptor levels in the kidney and coagulating gland, and lower levels in the hypothalamus and seminal vesicle when compred to C57BL/10J mice. B10D2F₁ mice had androgen receptor levels similar to those measured in C57BL/10J mice in all tissues studied, with the exception of the coagulating gland, where levels were similar to those observed in DBA/ZJ mice. These observations may indicate the existence of several loci coding for androgen receptors, with only one being expressed per tissue     

Strain difference in expression of the adult-type polycystic kidney disease gene, pcy, in the mouse

DBA/2FG-pcy and C57BL/6FG-pcy congenic strains were established by transferring the polycystic kidney disease gene, pcy, to DBA/2 and C57BL/6 mice. We carried out pathological and hematological examinations of these strains at 4, 8, 16 and 30 weeks of age. In DBA/2FG-pcy mice more than 8 weeks of age, macroscopic renal cysts were observed on the surface of both kidneys. Their kidneys weight was significantly greater than in DBA/2 mice at all ages examined. Microscopic renal cysts were evenly distributed at 4, 8 and 16 weeks of age. At 30 weeks of age, the kidneys were filled with numerous polycysts. In C57BL/6FG-pcy mice, no macroscopic renal cysts were found until the animals were 30 weeks old, and the weight of their kidneys was greater than in B6 mice of the same age. From 8 weeks of age on, a limited number of microscopic renal cysts was observed, and many renal cysts were found adjacent to the enlarged Bowman's capsules. With age, the red blood cell count and hematocrit level decreased while the platelet count increased in both strains, with greater changes occurring in DBA/2FG-pcy mice than in C57 BL/6FG-pcy mice. These findings demonstrate that polycystic kidney disease exhibits strain differences in animals with a DBA/2 and C57BL/6 background. Our results suggest that phenotypic expression of the pcy gene in the mouse depends on genetic background, and that variations in the severity of human polycystic kidney disease may be explained, at least in part, by individual differences in genetic background.     

Behavioral, physiological, and molecular differences in response to dietary restriction in three inbred mouse strains

Food restriction paradigms are widely used in animal studies to investigate systems involved in energy regulation. We have observed behavioral, physiological, and molecular differences in response to food restriction in three inbred mouse strains, C57BL/6J, A/J, and DBA/2J. These are the progenitors of chromosome substitution and recombinant inbred mouse strains used for mapping complex traits. DBA/2J and A/J mice increased their locomotor activity during food restriction, and both displayed a decrease in body temperature, but the decrease was significantly larger in DBA/2J compared with A/J mice. C57BL/6J mice did not increase their locomotor activity and displayed a large decrease in their body temperature. The large decline in body temperature during food restriction in DBA/2J and C57BL/6J strains was associated with a robust reduction in plasma leptin levels. DBA/2J mice showed a marked decrease in white and brown adipose tissue masses and an upregulation of the antithermogenic hypothalamic neuropeptide Y Y(1) receptor. In contrast, A/J mice showed a reduction in body temperature to a lesser extent that may be explained by downregulation of the thermogenic melanocortin 3 receptor and by behavioral thermoregulation as a consequence of their increased locomotor activity. These data indicate that genetic background is an important parameter in controlling an animal's adaptation strategy in response to food restriction. Therefore, mouse genetic mapping populations based on these progenitor lines are highly valuable for investigating mechanisms underlying strain-dependent differences in behavioral physiology that are seen during reduced food availability.     

Comparison of urethane-induced sister-chromatid exchanges in various murine strains, and the effect of enzyme inducers

The induction of sister-chromatid exchanges (SCEs) by urethane, 150 and 300 mg/kg administered i.p., was examined in bone-marrow cells of AKR, BALB/c, C3Hf, C57BL/6J and DBA/2 male mice. In all strains, the base-line level of SCE/cell was similar, ranging from 4.3 to 8.7, and the response increased with the dose of urethane. DBA/2 mice were the most susceptible to urethane at both dose levels, with 30.6 SCE/cell after treatment with 300 mg/kg, whereas the response of the other strains was from 17.4 to 21.5 SCE/cell at the same urethane dose. Pretreatment of C57BL/6J and DBA/2 mice with phenobarbital decreased the SCE frequencies induced by urethane, 300 mg/kg, to 70%, whereas a prior administration of beta-naphthoflavone reduced SCE levels in C57BL/6J but not in DBA/2 mice.     

Plasmodium chabaudi chabaudi (AS): differential cellular responses to infection in resistant and susceptible mice

The infection with blood stages of Plasmodium chabaudi chabaudi (AS) was followed in BALB/c and DBA/2 mice. Both strains show a peak parasitemia by 7-9 days after infection, display splenic hypercellularity of T and B cells, thymic atrophy, nearly complete depletion of B cells in the bone marrow, and mount comparable polyclonal IgM and IgG responses in the serum. In contrast, these strains diverge in some aspects of the immune response and susceptibility to infection: while BALB/c survive, 70-80% of DBA/2 die within 2 weeks; BALB/c but not DBA/2 show marked increases in the levels of splenic gamma/delta and regulatory T cells, dendritic cells and macrophages and parasite-specific IgM and IgG levels; however, lower levels of TNF-alpha and IL-12 were observed. These results suggest the relevance of different cell populations that are known to participate/regulate specific antibody responses and cytokine production in the susceptibility to infection.     

Observations on soft tissue calcification in DBA/2NCrj mice in comparison with CRJ:CD-1 mice

Male and female DBA/2NCrj (DBA/2) mice 3, 4, 5 and 10 weeks old were examined biochemically and pathologically and the results obtained were compared with those for CRJ:CD-1 (ICR) mice of the same age. The plasma levels of glucose, triglyceride and total cholesterol tended to be lower in DBA/2 mice than in ICR mice but the levels of non-esterified fatty acid, calcium and inorganic phosphorus were almost the same in the two strains. The mean body weight of DBA/2 mice was significantly lower than that of ICR mice at each examination, and the relative weights of the hearts of male and female DBA/2 mice were significantly greater than those of male and female ICR mice. Cardiac calcinosis, tongue calcification and corneal degeneration occurred exclusively in DBA/2 mice with incidences of 30%-100%. The incidence and severity of these lesions increased with age but no sex differences were seen. It was difficult to relate differences in biochemical features of the two strains with pathological findings obtained in the DBA/2 mice. The numbers of cells secreting adrenocorticotropic hormone in the pituitary glands were significantly greater in male and female DBA/2 mice than in ICR mice, suggesting a higher secretion of corticosteroids in the former strain.     

IL-4-producing gamma delta T cells that express a very restricted TCR repertoire are preferentially localized in liver and spleen.

IL-4-producing gamma delta thymocytes in normal mice belong to a distinct subset of gamma delta T cells characterized by low expression of Thy-1. This gamma delta thymocyte subset shares a number of phenotypic and functional properties with the NK T cell population. Thy-1dull gamma delta thymocytes in DBA/2 mice express a restricted repertoire of TCRs that are composed of the V gamma 1 gene product mainly associated with the V delta 6.4 chain and exhibit limited junctional sequence diversity. Using mice transgenic for a rearranged V gamma 1J gamma 4C gamma 4 chain and a novel mAb (9D3) specific for the V delta 6.3 and V delta 6.4 murine TCR delta chains, we have analyzed the peripheral localization and functional properties of gamma delta T cells displaying a similarly restricted TCR repertoire. In transgenic mice, IL-4 production by peripheral gamma delta T cells was confined to the gamma delta+9D3+ subset, which contains cells with a TCR repertoire similar to that found in Thy-1dull gamma delta thymocytes. In normal DBA/2 mice such cells represent close to half of the gamma delta T cells present in the liver and around 20% of the splenic gamma delta T cells.     

Emotionality, exploratory behavior, and locomotion in aging inbred strains of mice.

Two inbred strains of mice, C57BL/6J and DBA/2J, ranging in age from 2 to 38 months, were tested in an open field using the free exploration method. Scores were obtained for locomotor activity, exploratory behavior and emotionality. Strain differences were observed for all three variables. Beginning at late maturity (12 months), locomotor activity decreased with increasing age. Exploratory behavior was at a low level for DBA/2J mice at all ages. For C57BL/6J mice, exploratory behavior decreased significantly between 2 and 6 months and remained stable thereafter. Emotionality remained unchanged with advancing age for both strains of 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.     

Differences in expression of cell surface co-stimulatory molecules, Toll-like receptor genes and secretion of IL-12 by bone marrow-derived dendritic cells from susceptible and resistant mouse strains in response to Coccidioides posadasii

Coccidioides posadasii is a soil fungus that causes coccidioidomycosis or Valley Fever in the endemic regions of the southwestern US and Central America. Persons with decreased T cells reactivity and immune deficiency are at increased risk of developing severe disseminated infection. Among different mouse strains, DBA/2 mice are relatively resistant to C. posadasii whereas BALB/c mice are highly susceptible, and this discrepancy has been attributed to the difference in the development and expression of their Th1 cellular response. Dendritic cells (DC) are the most potent antigen-presenting cells that are activated after taking up pathogens or pathogens-derived antigens and regulate the immune response in the host, including Th1 cellular response. However, the DC responses against C. posadasii are not characterized. In the present study, we cultured bone-marrow derived DC (BMDC) from BALB/c and DBA/2 mice and infected with C. posadasii arthroconidia. The activation of BMDC was characterized by studying expression of cell surface co-stimulatory molecules (CD11c, MHC class II, CD40, CD80, and CD86), expression of genes encoding Toll-like receptors and release of IL-12. We found that the BMDC from DBA/2 mice showed significant upregulation of Toll-like receptor-2 and 4 genes expression, secretion of IL-12 (p<0.05) and modest increase in T cell co-stimulatory molecules as compared to BMDC from BALB/c mice. The data suggest that the differences in the activation status of DC in DBA/2 and BALB/c mice may be responsible for the discrepancy in their susceptibility to C. posadasii.     

The DBA.2 mouse is susceptible to disease following infection with a broad, but limited, range of influenza A and B viruses

We assessed the relative susceptibilities to disease of the DBA.2 and C57BL/6 mouse models upon infection with a range of influenza A and B viruses. DBA.2 mice were more susceptible to disease upon inoculation with human H1N1 influenza A virus strains, several swine influenza viruses, and influenza B viruses but were not overtly susceptible to infection with human seasonal H3N2 strains. Hemagglutination inhibition and immunoglobulin isotype profiling indicated that DBA.2 and C57BL/6 mice generate comparable humoral responses upon equivalent 50% mouse lethal dose (MLD(50)) challenges with influenza virus. Our data demonstrate the utility of DBA.2 mice for the elucidation of influenza virus pathogenicity determinants and the testing of influenza vaccines.     

Dimethylnitrosamine metabolism: II. In vitro activation of dimethylnitrosamine by hepatic and renal tissues from crosses among BALB/c, DBA and C57BL mice

Crosses among BALB/c, C57BL and DBA mice were performed to investigate the genetic mechanisms involved in metabolism of DMN by renal and hepatic tissues. Liver S-9 fractions from parental strain DBA had the greatest potential to activate DMN and liver fractions from parental strain BALB/c had the lowest. No age or sex-related differences were observed within strain. Crossing of either C57BL or DBA to BALB/c mice resulted in F1 hybrids with liver microsomal enzymes that gave results similar to the BALB/c parental strain. There were no sex or age differences within crossbred strains in the potential of liver to activate DMN. In contrast male DBA and C57BL parental mice renal S-9 fractions did not differ significantly from each other but did differ significantly from male BALB/c renal fractions and from female and immature animals of all strains. Crossing of either DBA or C57BL mice with BALB/c mice resulted in male F1 hybrids whose renal S-9 fractions did not differ significantly from males of the parental BALB/c strain. In all instances, male renal S-9 fractions had a significantly greater potential to activate DMN than female or immature animals. F1 DBA X C57BL hybrids had renal S-9 fractions that did not differ significantly from the parental strains. These data suggest that the gene(s) for low DMN metabolism of BALB/c mice are apparently dominant over the genes from both DBA and C57BL. The exact genetic or physiological mechanism needs further elucidation.     

The role of dopamine receptors in controlling mouse aggressivity: the genotype dependence]

Significant genotypic differences in shock-induced aggression were found in mice of eight inbred strains. Aggression was evaluated in test with the action of low electric current through the cage floor. Low aggressive strains C3H/He, DD, BALB/c, AKR and highly aggressive strains CBA, DBA/2. CC57Br were singled out by the number of aggressive attacks. Selective stimulation of dopamine D2 receptors by bromocriptine considerably increased the shock-induced aggressiveness in mice of low-aggressive strains. Blockade of D2-receptors by the injection of antagonist sulpiride decreased or prevented the manifestation of aggression in highly-aggressive mice. At the same time selective agonist of dopamine (D1) receptors SKF 38393 and administration of selective antagonist of D1-receptors SCH 23390 did not influence significantly shock-induced aggression. Thus, shock-induced aggression, depends on the animal genotype and activation of D2-receptors.     

Detection of Mls-antigens in various mouse strains using a new method

Mice of strains CBA and BALB/c, when injected with lymphocytes from theH-2-compatible Mls-antigen-incompatible strains C3H and DBA/2, respectively, develop a reduced lymphocyte reactivity against cells of the injected strains as measured in the mixed lymphocyte culture (MLC). The mechanism of the development of a depression of the MLC response against Mlsantigens is unknown. In this investigation we have tested the MLC response of lymphocytes from CBA mice preinjected with C3H lymphocytes against cells from 12 different strains. It was observed that the response decreased against cells from strains C3H, AKR, and A/Sn. Infusion of CBA mice with AKR lymphocytes decreased their MLC response against the same three strains. In contrast, infusion of CBA mice with A/Sn lymphocytes reduced their MLC responses against strains C3H, DBA/2, and the congenic strains A/Sn, A.SW, A.CA, and A.BY. BALB/c mice which were infused with DBA/2 lymphocytes developed reduced responses against DBA/2, C3H, and AKR. On the basis of these results we propose that mice of our strains C3H and AKR possess a common Mls-antigen which is strongly stimulatory, and that DBA/2 mice possess a second Mls-antigen which is also strongly stimulatory. The congenic strains A/Sn, A.SW, A.CA, and A.BY, which have differentH-2 complexes, possess a third Mls-antigen which is less stimulatory. The Mls-antigens of the strains listed above seem to exhibit extensive immunological crossreactivity.     

Strain Differences in Presynaptic Function: PROTEOMICS,ULTRASTRUCTURE, AND PHYSIOLOGY OF HIPPOCAMPAL SYNAPSES IN DBA/2J AND C57Bl/6J MICE

The inbred strains C57BL/6J and DBA/2J (DBA) display striking differences in a number of behavioral tasks depending on hippocampal function, such as contextual memory. Historically, this has been explained through differences in postsynaptic protein expression underlying synaptic transmission and plasticity. We measured the synaptic hippocampal protein content (iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) and mass spectrometry), CA1 synapse ultrastructural morphology, and synaptic functioning in adult C57BL/6J and DBA mice. DBA mice showed a prominent decrease in the Ras-GAP calcium-sensing protein RASAL1. Furthermore, expression of several presynaptic markers involved in exocytosis, such as syntaxin (Stx1b), Ras-related proteins (Rab3a/c), and rabphilin (Rph3a), was reduced. Ultrastructural analysis of CA1 hippocampal synapses showed a significantly lower number of synaptic vesicles and presynaptic cluster size in DBA mice, without changes in postsynaptic density or active zone. In line with this compromised presynaptic morphological and molecular phenotype in DBA mice, we found significantly lower paired-pulse facilitation and enhanced short term depression of glutamatergic synapses, indicating a difference in transmitter release and/or refilling mechanisms. Taken together, our data suggest that in addition to strain-specific postsynaptic differences, the change in dynamic properties of presynaptic transmitter release may underlie compromised synaptic processing related to cognitive functioning in DBA mice.     

Strain specificity and cholinergic modulation of visuospatial attention in three inbred mouse strains

The tremendous increase in the use of mouse inbred strains and mutant mice to study the molecular basis of psychiatric disorders urges for a better understanding of attentional performance in mice. To this aim, we investigated possible strain differences in performance and cholinergic modulation of visuospatial attention in three widely used mouse inbred strains (129S2/SvHsd, C57BL/6JOlaHsd and DBA/2OlaHsd) in the five-choice serial reaction time task. Results indicated that after extended training, performance of 129S2/SvHsd mice was superior to that of C57BL/6JOlaHsd and DBA/2OlaHsd mice in terms of attention, omission errors, inhibitory control and latencies to correct choice. Increasing the attentional load resulted in comparable decrements in attention in all strains and inhibitory control impairments that were most pronounced in DBA/2OlaHsd mice. Further pharmacological evaluation indicated that all strains showed attentional impairments after treatment with the muscarinic and nicotinic antagonists scopolamine and mecamylamine, respectively. 129S2/SvHsd mice were less sensitive, whereas DBA/2OlaHsd mice appeared more sensitive to the detrimental effects of mecamylamine. In addition, subchronic, but not acute, nicotine treatment slightly improved attentional performance in all strains to the same extent. In conclusion, our data indicate strain specificity with particularly good performance of 129S2/SvHsd mice and strong cholinergic involvement in visuospatial attention in mice.     

Bacillary growth, interleukin 2 production defect, and specific antibody secretion governed by different genetical factors in mice infected subcutaneously with Mycobacterium lepraemurium

Different mouse strains were infected subcutaneously in the footpad with 10(7) Mycobacterium lepraemurium (MLM). At various stages of the infection, the number of acid-fast bacilli (AFB) in different organs, spleen cell interleukin 2 production, and specific IgM and IgG serum antibodies to MLM sonicate were assessed. Strains were separable into two distinct groups depending on the number of AFB recovered from the different organs, without any obvious influence of the Bcg gene. Thus C57BL/6, DBA/2, (C57BL/6 X DBA/2)F1 and C3H/Pas mice belonged to the high resistance group and DBA/1, BALB/c, and CBA strains to the low resistance group. Interleukin 2 production was depressed only in C57BL/6 and C3H/Pas mice. Anti-MLM antibody response also markedly varied according to strains, in terms of antibody titers, Ig class distribution, and species specificity, but with a different genetic pattern from that observed for MLM growth control.