Influences of inbreeding and genetics on telomere length in mice

We measured telomere lengths of blood leukocytes in several inbred and outbred mammalian species, using a telomere-specific fluorescent probe and flow cytometry. Humans, non-human primates, and three outbred populations of Peromyscus mice (Peromyscus leucopus, Peromyscus maniculatus, and Peromyscus polionotus) have short telomeres. Two common strains of laboratory mice, C57BL/6J and DBA/2J, have telomeres several times longer than most other mammals surveyed. Moreover, the two inbred laboratory mouse strains display significantly different telomere lengths, suggesting the existence of strain-specific genetic determinants. To further examine the effects of inbreeding, we studied three Peromyscus leucopus inbred lines (GS109, GS16A1, and GS16B), all derived from the outbred P. leucopus stock. Telomeres of all three inbred lines are significantly lengthened relative to outbred P. leucopus, and the three lines display strain-specific significantly different telomere lengths, much like the C57BL/6J and DBA/2J strains of M. musculus. To further characterize the genetic inheritance of telomere length, we carried out several crosses to obtain hybrid F₁ mice between parental strains displaying the phenotype of long and short telomeres. In all F₁ mice assayed, peripheral blood leukocyte telomere length was intermediate to that of the parents. Additionally, we generated F₂ mice from a cross of the (P. leucopus outbred × GS16B)F₁. Based on the distribution of telomere length in the F₂ population, we determined that more than five loci contribute to telomere length regulation in Peromyscus. We concluded that inbreeding, through unknown mechanisms, results in the elongation of telomeres, and that telomere length for a given species and/or sub-strain is genetically determined by multiple segregating loci.     

Responses to novelty in two inbred strains of mice and their F1 hybrids: Behaviour and brain biogenic amines

Responses to novelty of two inbred strains of mice, C57BL/6JOrl (C) and A/JOrl (A) and their reciprocal F₁ hybrids were recorded. The parental strains differed from each other: the C strain showed a preference for a novel environment and neophobic reactions to a novel object introduced into the familiar environment. The A mice were characterized by a preference for the familiar environment and lower neophobia towards the novel object. In addition, the C strain exhibited high and the A strain low locomotor activity. The reciprocal hybrids were identical in their major responses to the C strain. Neurochemical investigations indicated that the parental A strain showed a lower level of dopamine in the striatum and in the olfactory bulbs when compared with the C strain and the hybrid F₁ C × A.     

F1 hybrid resistance: Long-term systemic effects sensitive to irradiation and age

In contrast to the usual rapid growth of transplanted syngeneic marrow cells in spleens of lethally irradiated recipients, the growth of parental marrow cells from certain inbred strains of mice is resisted by their F_l hybrids, other strains or both. The full complexity of this well known natural resistance is demonstrated here by using three inbred strains and their three Fl hybrids in all parent-hybrid combinations of donor and recipient. A similar resistance to parental marrow grafts is reported here in W-anemic F₁ hybrid recipients that are cured and repopulated without irradiation. Rather than resistance to short-term growth in spleens, F₁-hybrid resistance to permanent repopulation of the entire hemopoietic system is studied here. This manifestation of hybrid resistance is radiosensitive and declines in recipients over the age of 12 months. Long-term hemopoietic repopulation is measured quantitatively by injecting mixtures of two marrow-cell types with distinguishable hemoglobins into stem-cell-deficient recipients. A very high degree of resistance is detected against WB but not 136 parental marrow when mixed with WBB6F₁ marrow and injected into WBB6F₁ recipients. Most, but not all, of this resistance to permanent, systemic repopulation is abrogated by irradiation of the recipients; it is also abrogated after they reach the age of 15 months. Mouse models of long-term hybrid resistance studied in the entire hemopoietic system may be particularly relevant for marrow transplantation in man, where the objective is long-term systemic repopulation.     

Trichinella spiralis: Genetics of worm expulsion in inbred and F1 mice infected with different worm doses

The nematode Trichinella spiralis is rejected from the intestine at a time that is characteristic for each inbred strain of mouse. Previous work (R. G. Bell et al. 1982a) had empirically identified strong, intermediate, and weak phenotypes (NFR, $\text{C}\text{H}\text{He}$, and $\text{C57}\text{10}$ mice, respectively) in mice infected with 400 muscle larvae. It is shown that this classification applies to another eight inbred strains: SWR, $\text{DBA}\text{2}$, $\text{DBA}\text{1}$, LP, $\text{Bub}\text{Bn}$—all intermediate, and $\text{NZB}\text{BIN}$, C57L, A, and Mus molossinus—all weak. This phenotypic classification consistently applies with infections of 400–800 muscle larvae. Below doses of 300 muscle larvae, the strain designation of phenotype does not consistently apply. By this it is meant that the relative rejection rate changes for certain strains so that eventually some strains that were strong (NFR) or intermediate (AKR) responders to 400 muscle larvae become weak responders to 50 muscle larvae. Other strains increase their relative rejection time (B10 · BR, B10 · Q) while many do not change (NFS, $\text{C}\text{3}\text{Heb}\text{Fe}$, $\text{DBA}\text{2}$, $\text{DBA}\text{1}$). The phenomenon is most apparent in inbred parental strains rather than in F₁ crosses, and it represents a phenotypic variation in rejection time that is dependent on dose. It is also demonstrated that time of rejection is directly proportional to dose in all inbred and F₁ mouse strains that we have examined. Analysis of F₁ crosses shows that most have the rejection time of the strongest responding parental line, suggesting simple genetic control of strong, intermediate, and weak responses. Two F₁ crosses invalidated this theory. The $\text{DBA}\text{1}\text{×}\text{C3}\text{H}\text{He}$ (intermediate × intermediate) showed a strong response. The additive effects of parental rejection phenotype indicated that these lines could not be genetically identical for intermediate responsiveness. Similarly, the NFR (strong) × B10 · BR (weak) F₁ showed intermediate rejection, indicating partial dominance of $\text{C}\text{57}\text{B}\text{1}\text{10}$ genes over the strong responder NFR strain. Neither the primary expulsion time phenotype, phenotypic variation to low doses, or the rejection characteristics of F₁ crosses could be ascribed to genes linked to the major histocompatibility complex.     

Genetic studies of murine catalase: Regulation of multiple molecular forms of kidney catalase

Starch gel electrophoresis of kidney catalase in inbred strains C3H and C57BL/6, their F₁ hybrid, and first and second backcross generations demonstrated that single-component (type A) v. multiple-component (type B) electrophoretic patterns are controlled by a single locus. The type A electrophoretic pattern is dominant. Twenty-five inbred strains of mice were classified according to their kidney catalase electrophoretic pattern. The data indicate that the segregating genetic factor determines a specific substance in the type A kidney which affects the electrophoretic mobility of catalase. A comparison of the F₁ hybrid enzyme with a 1:1 mixture of C3H and C57BL/6 enzyme showed that the alteration of electrophoretic mobility is the result of posttranslational modification of the catalase molecule. An association of kidney catalase electrophoretic pattern and the H-2 ^k haplotype indicates that the locus controlling the electrophoretic pattern is most likely located on chromosome 17 in close proximity to the H-2 complex.     

The genetic basis of non-disjunction: Increased incidence of hyperploidy in oocytes from F1 hybrid mice

Summary Oocytes from parental mice strains NMRI/Han, C57/bl and Balb/c and from F₁ hybrid lines were analysed for aneuploidy due to non-disjunction after gonadotropin-stimulated ovulation. No hyperploid oocytes were present in five of the strains studied. F₁ hybrids from crosses of NMRI/HanxC57/bl did ovulate, however, a significantly increased number of hyperploid oocytes, although females from their parental strains show a rather low incidence of non-disjunction. The evidence for a genetic basis for non-disjunction is assessed and possible causative factors are discussed.Dedicated to Professor Dr.P.E. Becker on the occasion of his 75th birthday     

Mating preferences of F2 segregants of crosses between MHC-congenic mouse strains

Male and female F₂ homozygotes from crosses between MHC-congenic inbred mouse strains were tested for MHC-associated mating preference. In three instances, of the four genotypic combinations so tested, marked MHC-associated mating preference was observed. This result greatly reduces the possibility that the observed mating preferences of MHC-congenic inbred strains can be explained wholly in terms of non-MHC genetic drift, or of residual non-MHC genetic disparity, or of fortuitous acquired strain characteristics unrelated to MHC. In two of the four combinations investigated, the MHC-related mating bias of F₂ segregants was similar to that of the genotypically similar inbred parent strains. In a third combination, F² segregants did not show the mating bias exhibited by the corresponding parent strains. In a fourth combination, F₂ segregants displayed an MHC-related mating bias that was evident in the corresponding parental inbred strains only when the colonies of the parent strains had been maintained in isolation from other strains. While the exhibition of mating preference by mice of the same genotypes may differ according to circumstances, as indicated, in no instance was preference reversed. Mating preference in a given combination of MHC genotypes, whenever it was observed, always favored the same MHC haplotype of the two alternative haplotypes represented. It appears that the familial MHC genotypes of mice and the environment in which the colonies are maintained influence their MHC-related mating preference, but it has yet to be decided whether these factors operate by determining exposure to particular MHC haplotypes.Abbreviations used in this paper are as follows B6 C57BL/6 - B10 C57BL/10 - BALE BALB/c - BALB.B BALB.B10 - INB inbred - MHC major histocompatibility complex See also Figure 1     

A quantitative genetic analysis of tissue-specific catalase activity inMus musculus

Tissue-specific catalase activity in 3-week-old animals from inbred mouse strains 129/ReJ, BALB/c, C3H/HeAnl/Cas-1^b, C3H/HeSnJ, C3H/S, C57BL/6J, and Swiss-Webster was found to be highly variable by analysis of variance (P=0.01). Appropriate crosses were made among strains which were classified as normal (BALB/c, C3H/HeSnJ, C3H/S), hypocatalasemic (129/ReJ, C57BL/6J), and acatalasemic (C3H/HeAnl/Cas-1^b) with respect to blood catalase activity to study the inheritance of the blood, kidney, liver, and lung catalase activity levels in a number of generations (reciprocal F₁'s, F₂, two backcrosses —BC₁ and BC₂— and some RI lines). Segregation analysis and statistical methods which tested different models of inheritance as well as calculations of heritability were used in an effort to assess and evaluate genetic parameters that affect catalase activity. Results indicate that the inheritance of blood catalase activity in the cross involving acatalasemic and normal (BALB/c, C3H/HeSnJ) strains is compatible with the single-locus difference between the parental strains; however, the difference between the acatalasemic and the hypocatalasemic strain (C57BL/6J) would require additional genetic interaction for a satisfactory explanation. A similar pattern of generalization also applies to the inheritance of kidney catalase activity. The segregation pattern for the liver and lung catalase activity in most crosses is significantly different from the expectations of the single locus model. These results are compatible with the concept that a number of genes must affect tissue-specific catalase activity in mice. These may include previously described (e.g., Ce-1 and Ce-2) or novel genetic regulators/modifiers which interact with a single structural gene (Cas-1) or its product to produce the catalase phenotype characteristic of specific tissues in each strain.This investigation was supported by a Natural Sciences and Engineering Research Council of Canada operating grant to S.M.S.     

Effects of ENU dosage on mouse strains

The germline supermutagen, N-ethyl-N-nitrosourea (ENU), has a variety of effects on mice. ENU is a toxin and carcinogen as well as a mutagen, and strains differ in their susceptibility to its effects. Therefore, it is necessary to determine an appropriate mutagenic, non-toxic dose of ENU for strains that are to be used in experiments. In order to provide some guidance, we have compiled data from a number of laboratories that have exposed male mice from inbred and non-inbred strains or their F₁ hybrids to ENU. The results show that most F₁ hybrid animals tolerate ENU well, but that inbred strains of mice vary in their longevity and in their ability to recover fertility after treatment with ENU. Received: 11 February 2000 / Accepted: 11 February 2000     

Mendelian inheritance of variations in β-galactosidase activities in the house mouse

A genetic test of differences in -galactosidase activities in three mouse tissues, liver, kidney, and spleen, is demonstrated. Activities fall in three distinct categories in F ₂ crosses between the two inbred strains C57BL/K1 and DBA/2/K1. C57 mice consistently show high activities in all three tissues, and DBA mice show low activities except for some male kidneys. F ₁ mice are intermediate to the parental strains. There seems to be a simple mendelian ratio 1:2:1, between the numbers of animals belonging to the three activity classes in F ₂ crosses and a 1:1 ratio in backcrosses. Thus it is suggested that one single locus is responsible for most of the differences seen in this system.This work was supported by the Nilsson-Ehle fund and the Marcus Borgström fund.     

Quantitative trait locus and haplotype mapping in closely related inbred strains identifies a locus for open field behavior

Quantitative trait locus (QTL) mapping in the mouse typically utilizes inbred strains that exhibit significant genetic and phenotypic diversity. The development of dense SNP panels in a large number of inbred strains has eliminated the need to maximize genetic diversity in QTL studies as plenty of SNP markers are now available for almost any combination of strains. We conducted a QTL mapping experiment using both a backcross (N₂) and an intercross (F₂) between two genetically similar inbred mouse strains: C57BL/6J (B6) and C57L/J (C57). A set of additive QTLs for activity behaviors was identified on Chrs 1, 9, 13, and 15. We also identified additive QTLs for anxiety-related behaviors on Chrs 7, 9, and 16. A QTL on Chr 11 is sex-specific, and we revealed pairwise interactions between QTLs on Chrs 1 and 13 and Chrs 10 and 18. The Chr 9 activity QTL accounts for the largest amount of phenotypic variance and was not present in our recent analysis of a B6 × C58/J (C58) intercross (Bailey et al. in Genes Brain Behav 7:761–769, 2008). To narrow this QTL interval, we used a dense SNP haplotype map with over 7 million real and imputed SNP markers across 74 inbred mouse strains (Szatkiewicz et al. in Mamm Genome 19(3):199–208, 2008). Evaluation of shared and divergent haplotype blocks among B6, C57, and C58 strains narrowed the Chr 9 QTL interval considerably and highlights the utility of QTL mapping in closely related inbred strains.     

Genetic loci for diet-induced atherosclerotic lesions and plasma lipids in mice

Genetic factors independent of those affecting plasma lipid levels are a major contributor to risk for atherosclerosis in humans, yet the basis for these is poorly understood. This study examined plasma lipids and diet-induced atherosclerosis in 16-month-old female mice of strains C56BL/6J and DBA/2J. Mice of the parental strains, from recombinant inbred strains derived from these (BXD RI), and F₂ progeny were fed an atherogenic diet for 16 weeks, beginning at 1 year of age. This induced atherosclerotic lesion formation in both parental strains, accompanied by increased plasma LDL levels. However, individual BXD RI strains and the BXD F₂ mice demonstrated a range of atherosclerotic lesion formation that was not or at best weakly correlated with plasma lipid levels. Quantitative trait locus (QTL) analysis of the BXD F₂ mice identified a locus with significant linkage (lod 4.5) for aortic lesion size on Chromosome (Chr) 10 that was independent of plasma lipids. Other loci with suggestive or significant linkage for various plasma lipid measures were identified on Chr 2, 3, 4, 5, 6, 7, 11, and 17. In this intercross, the genes primarily influencing atherosclerosis are distinct from those controlling plasma lipid levels.     

Control of UVB immunosuppression in the mouse by autosomal and sex-linked genes

Irradiation with UVB (290–320 nm) initiates a systemic immunosuppression detectable as suppression of contact hypersensitivity (CHS). We investigated susceptibility to UV suppression in reciprocal F₁-hybrid and backcross mice derived from BALB/c (low susceptibility) and C57BL/6 (high susceptibility) inbred strains. CB6F₁ male mice exhibited high susceptibility and B6CF₁ male mice exhibited low susceptibility, indicating a major X-linked effect in the genetic control of UV immune suppression. Females of either F₁ hybrid showed intermediate suppression, consistent with random X-inactivation. A model of monogenic X-linked control was not sufficient, and evidence for the action of two genetically unlinked autosomal genes was found in parental backcross animals. Both sexes of (BALB/c × CB6F₁) mice showed a 1 high: 1 low ratio of phenotypes, indicating control by a major autosomal locus, Uvs1, confirmed by propagation of the high phenotype through selective backcrossing for nine generations to BALB/c. Uvs1 was not genetically linked to 12 chromosomal markers including the pigment genes b (brown) and c (albino). Backcross animals (C57BL/6 × CB6F₁) showed a significant sex difference, male mice giving a 3 high: 1 low ratio of phenotypes, compatible with the action of a second autosomal locus, Uvs2, in this hybrid. The findings are compatible with a model in which high phenotype (Uvs1 ^b/Uvs1 ^b) is dominant when subjected to recessive epistatis by the X-chromosome locus Uvs3, or by the autosomal locus Uvs2. The finding of genetic control by interacting autosomal and X-linked genes is unique. Genetically determined high susceptibility to UV immunosuppression may be an important risk factor for UV-related human diseases.     

Food neophobia in wild and laboratory mice (Mus musculus domesticus)

In a conditioned taste aversion procedure we were specifically interested in the topic of food neophobia. Wild and laboratory mice were individually presented with a novel drink (0.1 % saccharin solution). Compared with the daily water consumption, the intake of this was lower. This decrease was greater:(1) in wild than in tame populations ; (2) in random-bred (Swiss-albinos) than in inbred (C57 B1/6, BALB/c) strains ; (3) in F₁-hybrids (either wild x tame or inbred x inbred) than in the parental strains.These results are discussed:(1) in terms of a selective pressure linked to man's fight against rodents, leading to increased neophobia in wild mice ; and (2) by stressing the heterosis an inbreeding depression effects, which suggest that food neophobia is a component of Darwinian fitness.     

Phenotypic Plasticity Contributes to Maize Adaptation and Heterosis

Plant phenotypic plasticity describes altered phenotypic performance of an individual when grown in different environments. Exploring genetic architecture underlying plant plasticity variation may help mitigate the detrimental effects of a rapidly changing climate on agriculture, but little research has been done in this area to date. In the present study, we established a population of 976 maize F₁ hybrids by crossing 488 diverse inbred lines with two elite testers. Genome-wide association study identified hundreds of quantitative trait loci associated with phenotypic plasticity variation across diverse F₁ hybrids, the majority of which contributed very little variance, in accordance with the polygenic nature of these traits. We identified several quantitative trait locus regions that may have been selected during the tropical-temperate adaptation process. We also observed heterosis in terms of phenotypic plasticity, in addition to the traditional genetic value differences measured between hybrid and inbred lines, and the pattern of which was affected by genetic background. Our results demonstrate a landscape of phenotypic plasticity in maize, which will aid in the understanding of its genetic architecture, its contribution to adaptation and heterosis, and how it may be exploited for future maize breeding in a rapidly changing environment.     

Basal serum immunoglobulin levels

Two congenic strains of mice were identified that differ in their serum immunoglobulin levels. The strains were crossed, the F₁ progeny were intercrossed, and the serum immunoglobulin levels of the F₁ and F₂ progeny were analyzed. The F₁ mice have serum immunoglobulin levels like that of the high parent, and the low-immunoglobulin phenotype segregates in the F₂ population. Six other inbred strains of mice were also characterized for basal serum levels of five classes of immunoglobulin.     

Mapping of a quantitative trait locus for morphine withdrawal severity

Chronic morphine exposure results in physical dependence, manifested by physical symptoms during naloxone-precipitated withdrawal. Jumping frequency is widely considered the most sensitive and reliable index of withdrawal intensity in mice. Inbred mouse strains surveyed for naloxone-precipitated withdrawal display large and significant strain differences in jumping frequency, including an approximately tenfold difference between C57BL/6 and 129P3 mice. In the present study, (B6 × 129)F₂ hybrid mice were given daily morphine injections for four days using an escalating dosing schedule, and naloxone-precipitated withdrawal on day 5 was measured. A full-genome scan for linkage to phenotypic data was performed using polymorphic microsatellite markers. Significant linkage was observed between withdrawal jumping frequencies and a 28 cM-wide region of Chromosome 1 (32–60 cM; peak at 51 cM), accounting for 20% of the overall phenotypic variance. Two other suggestive QTLs were found, on Chromosomes 5 and 10, and an additive model fitting all three loci accounted for 43% of the total variance. F₂ mice were also assessed for changes in morphine analgesic potency using the tail-withdrawal test in dose–response studies on days 1 and 4. No linkage was observed between Chromosomes 1, 5, and 10 and morphine analgesic tolerance, suggestive of genetic dissociation of naloxone-precipitated withdrawal from morphine and chronic morphine intake per se. The significant quantitative trait locus for naloxone-precipitated withdrawal severity in morphine-dependent mice, which we name Depmq1, may prove to be of considerable heuristic value once the underlying gene or genes are identified.     

Differentiation of adrenomedullary catecholamine synthesizing enzyme responses to repeated immobilization in hybrid rats

The response of adrenomedullary catecholamine synthesizing enzymes to repeated immobilization was studied in hybrid (F₁) offspring of 2 inbred rat strains (LEW and F344). Immobilization-induced increases in tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and phenylethanolamine-N-methyl-transferase (PNMT) activities in one of the parental strains (F344) previously were shown to be dependent upon intact adrenal gland innervation but independent of the pituitary gland, while responses in the other parental strain (LEW) were independent of adrenal innervation but dependent upon pituitary function. Factors determining immobilization-induced increases in adrenal enzymes of F₁ offspring were enzyme-specific. Increased PNMT activity was pituitary dependent in F₁ rats, whereas increased TH and DBH activities after immobilization were dependent upon an intact adrenal gland innervation. These results suggest that the factor(s) regulating PNMT responses are differentiable from those regulating TH and DBH responses. The results also indicate that analysis of PNMT responses to immobilization in backcross populations is feasible, and could indicate whether strain-specific response mechanisms are heritable.     

Blood pressure in 15 inbred mouse strains and its lack of relation with obesity and insulin resistance in the progeny of an NZO/HILtJ × C3H/HeJ intercross

We characterized the systolic and diastolic blood pressures of 10-week-old males from 15 inbred mouse strains and found that blood pressures among strains were continuously distributed and that strain C3H/HeJ had the lowest mean systolic and diastolic pressure (100.5 ± 3.2 and 66.8 ± 3.5 mmHg), and a strain with obesity and diabetes, NZO/HILtJ, had the highest (132.4 ± 3.1 and 86.6 ± 6.9 mmHg). To understand the relationship of blood pressure with insulin resistance and obesity, we produced F₁ and F₂ progeny from reciprocal crosses of NZO, the strain with obesity, diabetes, and high blood pressure, and the strain with the lowest blood pressures, C3H/HeJ. Mean systolic pressures of 10-week-old (NZO × C3H)F₁ and (C3H × NZO)F₁ males were similar to each other (114.9 ± 3.8 and 117.2 ± 5.0 mmHg) and were intermediate to those of the parental strains. Systolic pressure of F₂ males (n = 223) was distributed normally about the mean, suggesting that blood pressure is a polygenic trait. The body mass index (BMI) and plasma insulin levels of F₂ progeny correlated significantly and positively with plasma leptin levels, suggesting that obesity is associated with insulin resistance. In contrast, systolic pressure did not correlate with BMI, plasma leptin levels, and plasma insulin levels, suggesting that genes underlying the development of hypertension in this intercross are not associated with the development of obesity and insulin resistance. Our results demonstrate that the progeny of NZO and C3H intercrosses are a practical and powerful tool for identifying blood pressure genes and for understanding human polygenic hypertension.(Fumihiro Sugiyama) These authors contributed equally to this study.