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Background and aims In a previous work, a female-specific atherosclerosis risk locus on chromosome (Chr) 3 was identified in an intercross of atherosclerosis-resistant FVB and atherosclerosis-susceptible C57BL/6 (B6) mice on the LDL-receptor deficient ( Ldlr −/− ) background. It was the aim of the current study to identify causative genes at this locus. Methods We established a congenic mouse model, where FVB.Chr3 B6/B6 mice carried an 80 Mb interval of distal Chr3 on an otherwise FVB. Ldlr −/− background, to validate the Chr3 locus. Candidate genes were identified using genome-wide expression analyses. Differentially expressed genes were validated using quantitative PCRs in F0 and F2 mice and their functions were investigated in pathophysiologically relevant cells. Results Fine-mapping of the Chr3 locus revealed two overlapping, yet independent subloci for female atherosclerosis susceptibility: when transmitted by grandfathers to granddaughters, the B6 risk allele increased atherosclerosis and downregulated the expression of the secreted phospholipase Pla2g12a (2.6 and 2.2 fold, respectively); when inherited by grandmothers, the B6 risk allele induced vascular cell adhesion molecule 1 ( Vcam1 ). Down-regulation of Pla2g12a and up-regulation of Vcam1 were validated in female FVB.Chr3 B6/B6 congenic mice, which developed 2.5 greater atherosclerotic lesions compared to littermate controls ( p =0.039). Pla2g12a was highly expressed in aortic endothelial cells in vivo , and knocking-down Pla2g12a expression by RNAi in cultured vascular endothelial cells or macrophages increased their adhesion to ECs in vitro . Conclusions Our data establish Pla2g12a as an atheroprotective candidate gene in mice, where high expression levels in ECs and macrophages may limit the recruitment and accumulation of these cells in nascent atherosclerotic lesions. [ABSTRACT FROM AUTHOR]

Women are at an increased risk for developing affective disorders during times of hormonal flux, including menopause when the ovaries cease production of estrogen. However, while all women undergo menopause, not all develop an affective disorder. Increased vulnerability can result from genetic predisposition, environmental factors and gene by environment interactions. In order to investigate interactions between genetic background and estrogen depletion, we performed bilateral ovariectomy, a surgical procedure that results in estrogen depletion and is thought to model the post-menopausal state, in a genetically defined panel of 37 inbred mouse strains. Seventeen days post-ovariectomy, we assessed behavior in two standard rodent assays of anxiety- and depressive-like behavior, the open field and forced swim tests. We detected a significant interaction between ovariectomy and genetic background on anxiety-like behavior in the open field. No strain specific effects of ovariectomy were observed in the forced swim assay. However, we did observe significant strain effects for all behaviors in both the open field and forced swim tests. This study is the largest to date to look at the effects of ovariectomy on behavior and provides evidence that ovariectomy interacts with genetic background to alter anxiety-like behavior in an animal model of menopause. [ABSTRACT FROM AUTHOR]

Nitrous oxide (N 2 O)-induced antinociception in mice is dependent on the neuromodulator nitric oxide (NO). In contrast to C57BL/6J (B6) mice, DBA/2J (D2) mice fail to respond to N 2 O with a robust antinociceptive response or with an increase in brain nitric oxide synthase (NOS) enzyme activity, suggesting that failure of D2 mice to respond to N 2 O might result from a deficit of NO function. Therefore, it was of interest to determine whether increasing the availability of NO might increase sensitivity of D2 mice to N 2 O. Male D2 mice were pretreated with sub-antinociceptive intracerebroventricular doses of the NO donor 3-morpholinosydnoimine or the NO precursor l -arginine then assessed for responsiveness to N 2 O-induced antinociception using the acetic acid abdominal constriction test. Both pretreatments increased the antinociceptive responsiveness of D2 mice to N 2 O. These results indicate that the NOS enzyme in D2 mice is functional and that the deficit in NO function that obstructs sensitivity to N 2 O-induced antinociception may lie in availability or utilization of l -arginine. [ABSTRACT FROM AUTHOR]

Mounting evidence suggests the association of immune genes with complex neuropsychiatric diseases, such as schizophrenia. However, immune gene expression in the brain and their involvement in schizophrenia-related behavior in animal models have not been well studied so far. We analyzed the social (resident–intruder) and sensorimotor gating (pre-pulse inhibition (PPI) of acoustic startle) behaviors, and expression profiles of several brain immune genes in adult C57BL/6N and DBA/2J female mice. Compared to C57BL/6N mice, DBA/2J mice exhibited less social interaction in the resident–intruder test and reduced pre-pulse inhibition. The mRNA levels of Il1b and Il6 genes were significantly higher in the cortex and hypothalamus, while the mRNA level of C1qb was lower in the cortex, hippocampus and hypothalamus of DBA/2J mice compared to C57BL/6N mice. Furthermore, Tnfsf13b was up-regulated in the cortex and hippocampus, and so did Cd47 in the hippocampus, while Cx3cl1 was down-regulated in the cortex of DBA/2J mice. Our study demonstrates the differential expression of several immune genes in C57BL/6N and DBA/2J strains and more importantly provides clues on their potential importance in regulating schizophrenia-related endophenotypes in animal models. [ABSTRACT FROM AUTHOR]

Abstract: Animal models are widely used to gain insight into the role of genetics on bone structure and function. One of the main strategies to map the genes regulating specific traits is called quantitative trait loci (QTL) analysis, which generally requires a very large number of animals (often more than 1000) to reach statistical significance. QTL analysis for mechanical traits has been mainly based on experimental mechanical testing, which, in view of the large number of animals, is time consuming. Hence, the goal of the present work was to introduce an automated method for large-scale high-throughput quantification of the mechanical properties of murine femora. Specifically, our aims were, first, to develop and validate an automated method to quantify murine femoral bone stiffness. Second, to test its high-throughput capabilities on murine femora from a large genetic study, more specifically, femora from two growth hormone (GH) deficient inbred strains of mice (B6-lit/lit and C3.B6-lit/lit) and their first (F1) and second (F2) filial offsprings. Automated routines were developed to convert micro-computed tomography (micro-CT) images of femora into micro-finite element (micro-FE) models. The method was experimentally validated on femora from C57BL/6J and C3H/HeJ mice: for both inbred strains the micro-FE models closely matched the experimentally measured bone stiffness when using a single tissue modulus of 13.06GPa. The mechanical analysis of the entire dataset (n=1990) took approximately 44 CPU hours on a supercomputer. In conclusion, our approach, in combination with QTL analysis could help to locate genes directly involved in controlling bone mechanical competence. [Copyright &y& Elsevier]

Summary: Mice as a species are susceptible to tuberculosis infection while mouse inbred strains present wide spectrum of susceptibility/resistance to this infection. However, non-tuberculosis Mycobacterial infections usually cannot be modeled in mice of common inbred strains. Introduction of specific properties, such as gene mutations, recombinants, targeted gene knockouts significantly extended the use of mice to mimic human Mycobacterial infections, including non-tuberculosis ones. This review describes the available mouse models of tuberculosis and non-tuberculosis infections and drug therapy in these models. Mouse models of non-tuberculosis infections are significantly less developed than tuberculosis models, hampering the development of therapies. [ABSTRACT FROM AUTHOR]

Recent advances of induced pluripotent stem cells (iPSCs) has demonstrated that full development potential is closely related with the expression state of noncoding RNAs (ncRNAs) of the Dlk1- Dio3 imprinted gene cluster. However, few of them, especially the long noncoding RNAs (lncRNAs), have been characterized in detail. AK044800 is a transcript from the Dlk1- Dio3 imprinted region with little known information. This study reports original data on the expression pattern of AK044800 during embryogenesis. Expression analysis showed that AK044800 was specifically expressed in the brain at mid-gestation, E9.5 and E11.5. And at E15.5, its expression was mainly concentrated in the forebrain. In the late-gestation stage (E18.5), AK044800 expression was weaker in the brain and began to emerge in some other tissues during this period. Notably, the expression of AK044800 was biallelic in the brain, unlike other noncoding transcripts from this imprinted region. In addition, its expression was dependent on inbred mouse strains. This may be the first lncRNA that has been identified with a different expression between inbred mouse strains. This study may provide useful clues for further investigations of expression regulation and functions of lncRNAs of the Dlk1- Dio3 imprinted region. [ABSTRACT FROM AUTHOR]

Abstract: Intracerebroventricular administration of selective agonist of serotonin 5-HT7 receptor LP44 (4-[2-(methylthio)phenyl]-N-(1,2,3,4-tetrahydro-1-naphthalenyl)-1-pyperasinehexanamide hydrochloride; 10.3, 20.5 or 41.0 nmol) produced considerable hypothermic response in CBA/Lac mice. LP44-induced (20.5 nmol) hypothermia was significantly attenuated by the selective 5-HT7 receptor antagonist SB 269970 (16.1 fmol, i.c.v.) pretreatment. At the same time, intraperitoneal administration of LP44 in a wide range of doses 1.0, 2.0 or 10.0 mg/kg (2.0, 4.0, 20.0 μmol/kg) did not cause considerable hypothermic response. These findings indicate the implication of central, rather than peripheral 5-HT7 receptors in the regulation of hypothermia. The comparison of LP44-induced (20.5 nmol) hypothermic reaction in eight inbred mouse strains (DBA/2J, CBA/Lac, C57BL/6, BALB/c, ICR, AKR/J, C3H and Asn) was performed and a significant effect of genotype was found. In the same eight mouse strains, functional activity of 5-HT1A and 5-HT3 receptors was studied. The comparison of hypothermic responses produced by 5-HT7 receptor agonist LP44 (20.5 nmol, i.c.v.) and 5-HT1A receptor agonist 8-OH-DPAT 1.0 mg/kg, i.p. (3.0 μmol/kg), 5-HT3 receptor agonist m-CPBG (40.0 nmol, i.c.v.) did not reveal considerable interstrain correlations between 5-HT7 and 5-HT1A or 5-HT3 receptor-induced hypothermia. The selective 5-HT7 receptor antagonist SB 269970 (16.1 fmol, i.c.v.) failed to attenuate the hypothermic effect of 8-OH-DPAT 1.0 mg/kg, i.p. (3.0 μmol/kg) and m-CPBG (40.0 nmol, i.c.v.) indicating that the brain 5-HT7 receptor is not involved in the hypothermic effects of 8-OH-DPAT or m-CPBG. The obtained results suggest that the central 5-HT7 receptor plays an essential role in the mediation of thermoregulation independent of 5-HT1A and 5-HT3 receptors. [Copyright &y& Elsevier]

Abstract: Study objectives: (a) Develop a new statistical approach to describe the microarchitecture of wakefulness and sleep in mice; (b) evaluate differences among inbred strains in this microarchitecture; (c) compare results when data are scored in 4-s versus 10-s epochs. Design: Studies in male mice of four inbred strains: AJ, C57BL/6, DBA and PWD. EEG/EMG were recorded for 24h and scored independently in 4-s and 10-s epochs. Measurements and results: Distribution of bout durations of wakefulness, NREM and REM sleep in mice has two distinct components, i.e., short and longer bouts. This is described as a spike (short bouts) and slab (longer bouts) distribution, a particular type of mixture model. The distribution in any state depends on the state the mouse is transitioning from and can be characterized by three parameters: the number of such bouts conditional on the previous state, the size of the spike, and the average length of the slab. While conventional statistics such as time spent in state, average bout duration, and number of bouts show some differences between inbred strains, this new statistical approach reveals more major differences. The major difference between strains is their ability to sustain long bouts of NREM sleep or wakefulness. Scoring mouse sleep/wake in 4-s epochs offered little new information when using conventional metrics but did when evaluating the microarchitecture based on this new approach. Conclusions: Standard statistical approaches do not adequately characterize the microarchitecture of mouse behavioral state. Approaches based on a spike-and-slab provide a quantitative description. [ABSTRACT FROM AUTHOR]

Abstract: Standard anti-tuberculosis (TB) drug therapy had distinct effects on the bacilli burden in mice of DBA/2, C3H, SWR/J, and C57BL/6 inbred strains. To standardize the TB infection process, susceptible DBA/2 mice were infected with 1/10 of the dose used for relatively resistant C57BL/6 mice, such that the lung CFUs were roughly identical 3 weeks after infection when therapy was initiated. We found that TB treatment was more effective in the susceptible DBA/2 mice than in the relatively resistant C57BL/6 mice. [Copyright &y& Elsevier]

Abstract: Inbred mouse strains exhibit differences in susceptibility to influenza A infections. However, the molecular mechanisms underlying these differences are unknown. Therefore, we infected a highly susceptible mouse strain (DBA/2J) and a resistant strain (C57BL/6J) with influenza A H1N1 (PR8) and performed genome-wide expression analysis. We found genes expressed in lung epithelium that were specifically down-regulated in DBA/2J mice, whereas a cluster of genes on chromosome 3 was only down-regulated in C57BL/6J. In both mouse strains, chemokines, cytokines and interferon-response genes were up-regulated, indicating that the main innate immune defense pathways were activated. However, many immune response genes were up-regulated in DBA/2J much stronger than in C57BL/6J, and several immune response genes were exclusively regulated in DBA/2J. Thus, susceptible DBA/2J mice showed a hyper-inflammatory response. This response is similar to infections with highly pathogenic influenza virus and may serve as a paradigm for a hyper-inflammatory host response to influenza A virus. [Copyright &y& Elsevier]

Abstract: The selective agonist of serotonin 5-HT3 receptor 1-(3-chlorophenyl)biguanide hydrochloride (m-CPBG) administered intracerebroventricularly (40, 80 or 160nmol) produced long-lasting dose-dependent hypothermic response in AKR/2J mice. m-CPBG (160nmol i.c.v.) induced profound hypothermia (delta t =−4°C) that lasted up to 7h. m-CPBG (40nmol i.c.v.)-induced hypothermia was attenuated by 5-HT3 receptor antagonist ondansetron pretreatment. At the same time, intraperitoneal administration of m-CPBG in a wide range of doses (0.5, 1.0, 5.0 or 10.0mg/kg) did not affect the body temperature. These findings indicate: (1) the implication of central, rather than peripheral 5-HT3 receptor in the thermoregulation; (2) the inability of m-CPBG to cross blood–brain barrier in mice. The comparison of brain 5-HT3-induced hypothermic reaction in six inbred mouse strains (DBA/2J, CBA/Lac, C57BL/6, BALB/c, ICR, AKR/J) was performed and two highly sensitive to m-CPBG strains (CBA/Lac and C57BL/6) were found. In the same six mouse strains the functional activity of 5-HT1A receptor was studied. The comparison of hypothermic reactions produced by 5-HT1A receptor agonist 8-OH-DPAT (1.0mg/kg i.p.) and m-CPBG revealed significant correlation between 5-HT3 and 5-HT1A-induced hypothermia in five out of six investigated mouse strains. 5-HT1A receptor antagonist p-MPPI pretreatment (1mg/kg i.p.) diminished hypothermia produced by centrally administered m-CPBG (40nmol i.c.v.). The data suggest the cross-talk between 5-HT1A and 5-HT3 receptors in the mechanism of 5-HT-related hypothermia. [Copyright &y& Elsevier]

Abstract: Male mice from 14 standard inbred strains were exposed to morphine in a sustained released preparation injected subcutaneously. Five hours later withdrawal was precipitated by intraperitoneal injection of naloxone. Mice were tested from 0 to 15min after naloxone for withdrawal jumping behavior, and then from minute 15–16 for other signs, including boli count, presence of soft stool, lacrimation, “wet dog” shakes, and air chewing. They were also assessed for change in body temperature 17min after naloxone. Strains differed markedly in the severity of withdrawal for jumping, change in body temperature, and number of fecal boli. Strains also differed in percentage of animals displaying soft stool and air chewing behavior. The other two signs were seen at too low frequency for analysis. Correlations of strain mean withdrawal severity with other responses to morphine and other abused drugs showed that high morphine withdrawal jumping and low change in body temperature were both genetically related to high morphine consumption, but not generally to other measures of morphine withdrawal or morphine sensitivity. [Copyright &y& Elsevier]

Summary: Etiopathogenesis of depression and the cause of insensitivity to treatment remain poorly understood, although genetic makeup has been established as a contributing factor. The isogenicity of inbred mouse strains provides a useful tool for investigating the link between genes and behavior or drug response. Hence, our aim was to identify inbred mouse strains (among A/J, BALB/c, C3H, C57BL/6, CBA, DBA and FVB) sensitive to a 9-week period of unpredictable chronic mild stress (UCMS) and, from the fifth week onward, to the reversal effect of an antidepressant (AD) (imipramine, 20mg/kg/day i.p.) on various depression-related changes: physical, behavioral and neuroendocrine states. UCMS induced a significant deterioration of the coat state (in all the strains), blunted emotional reactivity in the novelty-suppressed feeding (NSF) test (A/J, BALB/c, C57BL/6), and changes in the level of fecal corticosterone metabolites (BALB/c, C57BL/6, DBA, FVB). Imipramine treatment reversed the UCMS-induced alterations of the coat state (BALB/c, DBA), in the NSF test (A/J, BALB/c, C57BL/6) and in fecal corticosterone metabolites (BALB/c, C57BL/6). C3H, CBA and FVB mice were irresponsive to imipramine treatment. It is noteworthy that UCMS-induced physical or behavioral changes occurred without hypothalamo–pituitary–adrenal (HPA) axis alterations in some strains (A/J, C3H, CBA), although the AD-induced reversal of these changes in BALB/c and C57BL/6 was associated with HPA axis normalization. Finally, UCMS is shown to discriminate various alterations and to replicate in a strain-dependent manner diverse profiles reminiscent of human disease subtypes. UCMS may thus enable the selection of strains suitable for investigating specific depression-related features and could be an appropriate model for identifying genetic factors associated with increased vulnerability, specific symptoms of affective disorders, and AD resistance. [Copyright &y& Elsevier]

Abstract: Complex corticocancellous skeletal sites such as the vertebra or proximal femur are connected networks of bone capable of transferring mechanical loads. Characterizing these structures as networks may allow us to quantify the load transferring behavior of the emergent system as a function of the connected cortical and trabecular components. By defining the relationship between certain physical bone traits and mechanical load transfer pathways, a clearer picture of the genetic determinants of skeletal fragility can be developed. We tested the hypothesis that the measures provided by network percolation theory will reveal that different combinations of cortical, trabecular, and compositional traits lead to significantly different load transfer pathways within the vertebral bodies among inbred mouse strains. Gross morphologic, micro-architectural, and compositional traits of L5 vertebrae from 15 week old A/J (A), C57BL6/J (B6), and C3H/HeJ (C3H) inbred mice (n =10/strain) were determined using micro-computed tomography. Measures included total cross-sectional area, bone volume fraction, trabecular number, thickness, spacing, cortical area, and tissue mineral density. Two-dimensional coronal sections were converted to network graphs with the cortical shell considered as one highly connected node. Percolation parameters including correlation length (average number of connected nodes between superior and inferior surfaces), chemical length (minimum number of connected nodes between surfaces), and backbone mass (strut number) were measured. Analysis of the topology of the connected bone networks showed that A and B6 mice transfer load through trabecular pathways in the middle of the vertebral body in addition to the cortical shell. C3H mice transfer load primarily through the highly mineralized cortical shell. Thus, the measures provided by percolation theory provide a quantitative approach to study how different combinations of cortical and trabecular traits lead to mechanically functional structures. The data further emphasize the interdependent nature of these physical bone traits suggesting similar genetic variants may affect both trabecular and cortical bone. Therefore, developing a network approach to study corticocancellous architecture during growth should further our understanding of the biological basis of skeletal fragility and, thus, provide novel engineering approaches to studying the genetic basis of fracture risk. [Copyright &y& Elsevier]

Abstract: Cochlear function was evaluated in a longitudinal study of 28 inbred strains of mice at 3 and 5 mo of age using measures of distortion product otoacoustic emissions (DPOAEs) in response to a federal initiative to develop rapid mouse phenotyping methodologies. DP-grams at f 2 frequencies ranging from 6.3 to 54.2kHz were obtained in about 3min/ear by eliciting 2f 1 − f 2 DPOAEs in 0.1-octave steps of f 2 with primary tones at L 1 = L 2 =55, 65, and 75dB SPL. CBA/CaJ mice exhibited average levels of ∼26dB SPL and this strain was selected as the normal reference strain against which the others were compared. Based upon the configurations of their DP-grams, the 28 mouse strains could be categorized into four distinct groups. That is, nine of the strains including the CBA were designated as the CBA-like group because these mice displayed robust DPOAE levels across frequency. In contrast, the remaining three groups all exhibited irregular DP-gram patterns. Specifically, eight of the remaining 19 strains showed a progressive high- to low-frequency reduction in DPOAE levels that was typical of age-related hearing loss (AHL) associated with mouse strains homozygous for the ahl allele and were labeled as AHL-like strains. Seven strains demonstrating relatively even patterns of reduced DPOAE levels across the frequency-test range were designated as Flat-loss strains. Finally, the remaining four strains exhibited no measurable DPOAEs at either 3 or 5 mo of age and thus were classified as Absent strains. Extending the f 2 test frequencies up to ∼54kHz led to the detection of very early-onset reductions in cochlear function in non-CBA-like groups so that all strains could be categorized by 3 mo of age. Predictably, the AHL-like strains showed more pronounced DPOAE losses at 5 mo than at 3 mo. A similar deterioration in DPOAE levels was not apparent for the Flat-loss strains. Both the AHL-like and Flat-loss strains showed considerably more variability in DPOAE levels than did the CBA-like strains. Together, these findings indicate that DP-grams adequately reveal both frequency-specific loss patterns and details of inbred strain variability. [Copyright &y& Elsevier]

Abstract: Excessive fluoride (F) can lead to abnormal bone biology. Numerous studies have focused on the anabolic action of F yet little is known regarding any action on osteoclastogenesis. Little is known regarding the influence of an individual''s genetic background on the responses of bone cells to F. Four-week old C57BL/6J (B6) and C3H/HeJ (C3H) female mice were treated with NaF in the drinking water (0 ppm, 50 ppm and 100 ppm F ion) for 3 weeks. Bone marrow cells were harvested for osteoclastogenesis and hematopoietic colony-forming cell assays. Sera were analyzed for biochemical and bone markers. Femurs, tibiae, and lumbar vertebrae were subjected to microCT analysis. Tibiae and femurs were subjected to histology and biomechanical testing, respectively. The results demonstrated new actions of F on osteoclastogenesis and hematopoietic cell differentiation. Strain-specific responses were observed. The anabolic action of F was favored in B6 mice exhibiting dose-dependent increases in serum ALP activity (p <0.001); in proximal tibia trabecular and vertebral BMD (tibia at 50&100 ppm, p =0.001; vertebrae at 50 and 100 ppm, p =0.023&0.019, respectively); and decrease in intact PTH and sRANKL (p =0.045 and p <0.001, respectively). F treatment in B6 mice also resulted in increased numbers of CFU-GEMM colonies (p =0.025). Strain-specific accumulations in bone [F] were observed. For C3H mice, dose-dependent increases were observed in osteoclast potential (p <0.001), in situ trabecular osteoclast number (p =0.007), hematopoietic colony forming units (CFU-GEMM: p <0.001, CFU-GM: p =0.006, CFU-M: p <0.001), and serum markers for osteoclastogenesis (intact PTH: p =0.004, RANKL: p =0.022, TRAP5b: p <0.001). A concordant decrease in serum OPG (p =0.005) was also observed. Fluoride treatment had no significant effects on bone morphology, BMD, and serum PYD cross-links in C3H suggesting a lack of significant bone resorption. Mechanical properties were also unaltered in C3H. In conclusion, short term F treatment at physiological levels has strain-specific effects in mice. The expected anabolic effects were observed in B6 and novel actions hallmarked by enhanced osteoclastogenesis shifts in hematopoietic cell differentiation in the C3H strain. [Copyright &y& Elsevier]

Abstract: Mouse models are widely used in genomic studies for quantitative trait loci (QTL) analyses. In the field of skeletal micro-structure, μCT has proven to be an invaluable imaging tool for the characterization of structural bone traits. However, the definition of analysis compartments requires a lot of user interaction, and therefore is not applicable as a standard way to analyze genetic linkage studies with several hundreds of animals. Here, we developed an automated three-dimensional based algorithm for a high-throughput regional analysis of three compartments in murine femora, including whole bone, cortical bone in the diaphysis and trabecular bone in the metaphysis. The algorithm relies on basic image processing concepts using morphological operators as well as a new approach of separating cortical from trabecular bone. Reproducibility of the automatic approach was investigated with respect to precision errors (PE %CV ) of micro-structural indices analyzed in these automatically defined compartments. The developed algorithm was then used to perform a high-throughput analysis of over 2000 femora in a genetic linkage study for further examination of stability and performance. Precision errors were 3.5% or less for all micro-structural indices. The analysis of one femur (mask generation and parameter evaluation) took 7 min on an AlphaServer DS25. The algorithm showed a very high reliability and worked successfully for 99.64% of all femora. Investigations of correlations amongst the assessed micro-structural indices together with heritability and polygene estimates revealed apparent volume density (AVD), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) and cortical thickness (Ct.Th) as candidates for a successful QTL analysis. The presented automatic analysis allows for standardized high-throughput phenotypic screening in mice femora for large genetic linkage studies with very high reliability and good precision. [Copyright &y& Elsevier]

Abstract: CYP1A2 expression is constitutively high in mouse liver and is well known for metabolizing several drugs and many procarcinogens to reactive intermediates that can cause toxicity or cancer. In the present study, the basal level of hepatic CYP1A2 activity was shown to vary among different inbred mouse strains. The highest methoxyresorufin-O-demethylase activity (261±52pmol/mgprotein/min) was registered in CC57BR and the lowest (82±11pmol/mgprotein/min) in C3H/a. We have tested the hypothesis that possible polymorphisms in regulatory elements in the 5′-upstream region of the mouse CYP1A2 gene could cause the differences in CYP1A2 enzyme activity among different inbred strains. We have performed a study on the CYP1A2 gene by sequencing the regulatory region from −4675 to −4204 where two enhancer elements were recently identified. The absence of mutation prescribing the phenotype in the CYP1A2 gene was found. The region studied seems to be a highly conserved in mice and not to be associated with interstrain differences in constitutive CYP1A2 enzyme activity. [Copyright &y& Elsevier]

Abstract: Introduction : The influence of genetic background on bone architecture and mechanical properties is well established. Nevertheless, to date, only few animal studies explore an underlying genetic basis for extrinsic factors effect such as fluoride effect on bone metabolism. Materials and methods : This study assessed the effect of increasing fluoride doses (0 ppm, 25 ppm, 50 ppm, 100 ppm) on the bone properties in 3 inbred mouse strains that demonstrate different susceptibilities to developing enamel fluorosis (A/J a “susceptible” strain, 129P3/J a “resistant” strain and SWR/J an “intermediate” strain). Fluoride concentrations were determined in femora and vertebral bodies. Bone mineral density was evaluating through DEXA. Finally, three-point bend testing of femora, compression testing of vertebral bodies and femoral neck-fracture testing were performed to evaluate mechanical properties. Results : Concordant with increasing fluoride dose were significant increases of fluoride concentration in femora and vertebral bodies from all 3 strains. Fluoride treatment had little effect on the bone mineral densities (BMD) in the 3 strains. Mechanical testing showed significant alterations in “bone quality” in the A/J strain, whereas moderate alterations in “bone quality” in the SWR/J strain and no effects in the 129P3/J strain were observed. Conclusion : The results suggest that genetic factors may contribute to the variation in bone response to fluoride exposure and that fluoride might affect bone properties without altering BMD. [Copyright &y& Elsevier]

Abstract: Neurological and psychiatric disorders are among the most common and most serious health problems in developed countries. Transgenic mouse models mimicking human neurological diseases have provided new insights into development and function of the nervous system. One of the prominent goals of the German National Genome Research Network is the understanding of the in vivo function of single genes and the pathophysiological and clinical consequences of respective mutations. The German Mouse Clinic (GMC) offers a high-throughput primary screen of genetically modified mouse models as well as an in-depth analysis in secondary and tertiary screens covering various fields of mouse physiology. Here we describe the phenotyping methods of the Neurological Screen in the GMC, exemplified in the four inbred mouse lines C57BL/6J, C3HeB/FeJ, BALB/cByJ, and 129S2/SvPas. For our primary screen, we generated “standard operating procedures” that were validated between different laboratories. The phenotyping of inbred strains already showed significant differences in various parameters, thus being a prerequisite for the examination of mutant mouse lines. [Copyright &y& Elsevier]

Abstract: We compared the behaviors of eight inbred mouse strains across 18 variables, using 11 behavioral assays, and gave ethanol (EtOH) as an intoxicant. Genetic influences on behavior and sensitivity to EtOH were pronounced, but strain sensitivities were generally only modestly correlated across tasks. Certain well-correlated clusters of responses suggested that some genes affect similar neurobiological substrates. No strains of mice were generally sensitive or resistant to intoxication across tasks. Anthropomorphically appealing concepts like ‘muscle strength’ had little explanatory power across tasks. A battery of selected tests was proposed for future studies. Overall, the results show that each mouse behavioral assay captures only a portion of ataxia, a genetically complex behavioral domain. Conversely, multiple behavioral capacities are apparently required for performance in each specific assay. Thus, if only one or two tests are used to evaluate motor function in genetically engineered mutant mice, only a small portion of the domain will be assessed and results may be misleading. This caveat likely extends to many behavioral domains (e.g. learning and memory, anxiety). [Copyright &y& Elsevier]

Abstract: The femurs from groups of mice from 29 different inbred strains were characterized to study the genetic variations in bone parameters. For these analyses, we used peripheral quantitative computed tomography to assess bone size and density in addition to three-point bend testing to assess bone mechanical properties. Highly significant differences between inbred strains were found for all size, density, and mechanical parameters measured (P &#60; 0.0001). Correcting femoral cross-sectional geometry values or bone mechanical properties values for body weight or femur length reduced but did not eliminate the variations in bone geometry or bone mechanical properties. Mice of similar body size had as much as a 40% difference in the midshaft total area of the femur. Regression analysis suggested that 50.9% of the variation in maximum load among strains was related to variations in section modulus, i.e., cross-sectional geometry, 21.5% was related to variations in material bone density, and 27.7% to variations in quality. These components were further analyzed to show that 3.9–27.8% of the variation in maximum load was related to adaptation to mechanical stress. These findings indicate that there is a significant genetic variation in the femur cross-sectional area, density, and mechanical properties between inbred mouse strains. These studies identify inbred mouse strains suitable for future studies identifying genes regulating bone geometry and mechanical properties. [Copyright &y& Elsevier]

Baseline distortion-product otoacoustic emissions (DPOAEs) at several primary-tone levels were compared between naive 2- to 3-month old inbred CBA/CaJ (CBA) and wild-derived MOLF/Ei (MOLF) mice. Only minor DPOAE differences were noted between the two strains and these differences were not systematic across frequency or test levels. These emission findings were consistent with earlier results on auditory brainstem response thresholds reported by others [Zheng et al., Hear. Res. 130 (1999) 94–107] thus suggesting that both CBA and MOLF strains have normal hearing. Subsequent episodes of over-exposure to a 105-dB SPL, octave-band noise centered at 10 kHz for 8 h revealed that MOLF DPOAEs were exceptionally resistant to the adverse aftereffects of excessive noise exposure as compared to CBA mice. Unlike the noise-exposure resistant inbred 129/SvEvTac strain, which has reduced baseline DPOAE levels especially at high frequencies, MOLF mice have normal DPOAEs making the interpretation of noise-exposure effects more straightforward. [Copyright &y& Elsevier]

Rationale: In a search for potential supplements or alternatives to the pharmacological treatment of epilepsy, we examined the effects of static magnetic fields on audiogenic seizures of DBA/2 mice. Methods: Two strains of DBA/2 mice were subjected to auditory stimulation that resulted sequentially in wild running, loss of righting, clonus, tonic hindlimb extension, and death in 80–95% of animals in different experiments. The incidence of seizure stages in groups of animals pretreated with a static magnetic field, phenytoin (PHT) or both was compared to the incidence in sham-exposed control mice. Results: Depending on magnetic flux density and duration of exposure to the field, seizure severity decreased significantly, but not completely, in both strains. However, incidence of five seizure stages was reduced in one strain, with about half of the mice seizure free. Two seizure stages (tonic hindlimb extension and death) were reduced significantly in the other. Magnetic field pretreatment potentiated the effect of PHT. Clonic seizures refractory to PHT or magnetic field pretreatment in DBA/2J mice responded to pretreatment with a combination of PHT and the magnetic field. Conclusions: A static magnetic field had some anticonvulsant effects when employed alone. More robust effects were seen in combination with PHT. Further testing of magnetic fields for anticonvulsant effects and elucidation of mechanisms of action seem to be warranted. [Copyright &y& Elsevier]

Recruitment of brite/beige cells, known as browning of white adipose tissue (WAT), is an efficient way to turn an energy-storing organ into an energy-dissipating one and may therefore be of therapeutic value in combating obesity. However, a comprehensive understanding of the regulatory mechanisms mediating WAT browning is still lacking. Here, we exploit the large natural variation in WAT browning propensity between inbred mouse strains to gain an inclusive view of the core regulatory network coordinating this cellular process. Combining comparative transcriptomics, perturbation-based validations, and gene network analyses, we present a comprehensive gene regulatory network of inguinal WAT browning, revealing up to four distinct regulatory modules with key roles for uncovered transcriptional factors, while also providing deep insights into the genetic architecture of brite adipogenesis. The presented findings therefore greatly increase our understanding of the molecular drivers mediating the intriguing cellular heterogeneity and plasticity of adipose tissue. • The browning capacity of white adipose tissue is under complex genetic control • Genetic architecture of brite adipogenesis is built from several genetic models • Dozens of Ucp1 regulating factors are validated • A core browning regulatory network with four distinct regulatory modules is revealed Browning of white fat, which turns an energy-storing organ into an energy-dissipating one, holds promise for the development of novel therapeutics against obesity. By exploiting intrinsic mouse strain variation and applying a systems-genetics approach, Li et al. reveal a comprehensive gene regulatory network that controls this browning process. [ABSTRACT FROM AUTHOR]