Behavioural studies in different strains of mice and the problem of heterosis

Significant differences between strains were found for open field activity and emotionality, exploratory activity, initial reaction to shock, conditioned avoidance learning, and weight for three inbred strains (Balb/c, C57Bl and C3H), such that the heaviest strain (Balb/c) was the least active and most emotional in the open field, gave the lowest score for exploratory activity and longest reaction to shock times, and was the poorest at conditioned avoidance learning.The hybrids from crosses with C57Bl tended more towards complete dominance and overdominance than did the Balb/c × C3H hybrid, probably because the Balb/c and C3H strains have some common ancestry.Heterosis was most marked for traits involving exploration or learning. Variability of the hybrids for these traits was often lower than the parents, i.e. they showed behavioural homeostasis. For other traits variability of the hybrids tended towards the parent whose genotype was dominant for the trait.Of the non-genetic factors studied, age and litter size were found to have the greatest effect on the behavioural traits. Parity was found only to affect weight. Sex differences were found for weight and emotionality.     

4-way bred rats as experimental animals

The present study is an attempt to utilize hybrids among several inbred strains of rats as useful animals for the studies of effectiveness and toxicology on drugs., Four-way crosses were made among the LEW, WM, F344 and DRY strains of rats, and their characteristics were examined. From the breeding data of diallel crosses among these four strains and reciprocal crosses among their F1 hybrids, the mating type indicating the highest reproductivity was (LEW X WM) F1 X (F344 X DRY) F1. These four-way crosses were designated as LWFD. The reproductivity of this mating type was exceedingly higher than those of four strains. In order to examine the susceptibility to thiamine hydrochloride, the acute toxicity test was practiced in inbred strains, F1 hybrids and four-way crosses. As a result, in spite of highly heterogeneous population, the LWFD did not show a peculiar response in comparison with four strains and their F1 hybrids. Furthermore, hematological and clinico-biochemical values of the LWFD did not show a large variability as presumed. From these results, it is suggested that hybrids such as four-way crosses among inbred strains can be used as useful animals for the studies of effectiveness and toxicology on drugs.     

γ-Glutamyl Cyclotransferase: A New Genetic Polymorphism in the Mouse (MUS MUSCULUS) Linked to LYT–2

Electrophoretically variant forms of gamma-glutamyl cyclotransferase have been identified in red cells of inbred mouse strains. Each inbred strain exhibited a major band of activity and a minor band that migrated more anodally. The polymorphism affects the migration of both the major and minor bands in a similar way. F1 hybrids between strains with fast forms (A/J) and strains with the slow forms (C57BL/6J) exhibited a four-banded pattern consistent with co-dominant inheritance. The patterns observed in backcross and F2 mice were consistent with the segregation of a pair of autosomal co-dominant alleles. Recombinant inbred strains and a congenic strain were used to show that the locus controlling gamma-glutamyl cyclotransferase (Ggc) is linked to Lyt-2, a lymphocyte alloantigen locus on chromosome 6, with an estimated map distance of 5.0 +/- 2.5 centimorgans.     

Characterization of silver-stained nucleolus organizer regions (Ag-NORs) in 16 inbred strains of the Norway rat, Rattus norvegicus

Silver-stained nucleolus organizer regions (Ag-NORs) in metaphase chromosomes of cultured fibroblasts were compared among 16 inbred strains of the Norway rat, Rattus norvegicus. Ag-NORs were located at the secondary constrictions or juxtacentromeric regions of the short arms of chromosomes 3, 11, and 12. The frequency and relative size of Ag-NORs were found to be strain-specific, providing a genetic marker system useful for characterization of inbred strains. While considerable cell-to-cell variation was observed within a given strain, the strain-specific pattern of Ag-NORs was shown to be consistent in cultured and noncultured cells obtained from different tissues of embryos, newborns, and adults, as well as in successively cultured cells examined up to the 10th subculture generation. The patterns of Ag-NORs in F1 hybrids made between some of the inbred strains were in general agreement with those expected from the parental strains; some unexpected patterns were noted in F1 hybrids of a particular cross, suggesting the possible existence of nucleolar interactions in such interstrain hybrids, although this has to be confirmed.     

Inheritance of dermatoglyphic configurations in the rat

Inheritance of dermatoglyphic configurations was studied on the palmar III interdigital pad of the rat (Rattus norvegicus). Four rats from each of the WKS and ACI/N inbred strains were mated with each other, and 54 F1 and 88 F2 hybrids were obtained. In addition, 52 offspring were produced by backcross mating between F1 hybrids and WKS and ACI/N parents. Whorls were the predominant pattern in the WKS strain and triradial patterns characterized the ACI/N strains. The F1 hybrids showed a wide range of pattern types, consisting of whorls, loops, cusps, arches, and triradial patterns. These patterns were intermediate in size between the two inbred strains. The F2 hybrids presented a distribution of patterns with a similar range as the F1, but the frequencies of some types were different. Patterns in the offspring of each backcross demonstrated a slight shift towards the characteristic pattern of the parental inbred strain. No sex difference was observed. Generally, the bilateral differences were striking, with a radial direction predominating on the left palm, and an ulnar one on the right palm, respectively. This study suggests that the dermal patterns are genetically determined, but also are influenced by environmental factors, especially in the hybrids.     

Paternal inheritance of egg traits in mice: a case of genomic imprinting

Eggs from reciprocal hybrids between the C57BL/6By and BALB/cBy strains were tested for their susceptibility to attack by hyaluronidase and pronase. There were significant reciprocal differences between the F1 females in the responses of their unfertilized eggs to both enzymes. The F1 hybrids from BALB mothers showed the increased susceptibility characteristic of C57BL whilst the F1 hybrids with C57BL mothers were more resistant to both enzymes, like BALB mice. Eggs from the four kinds of reciprocal F2 hybrid females also showed patroclinous patterns of susceptibility. A patroclinous difference was found between reciprocal crosses of the CXBD and CXBE recombinant inbred strains but not in crosses between recombinant inbred strains with similar phenotypes. Cross fostering did not alter the phenotypes of the C57BL and BALB females or those of their reciprocal F1 hybrids. The findings are interpreted in terms of differential genomic imprinting of paternally inherited information. The possible general usefulness of patroclinous differences between reciprocal F1 females in revealing differences in imprinting is noted.     

Allelic expression changes in Medaka (Oryzias latipes) hybrids between inbred strains derived from genetically distant populations

Variations in allele expressions between genetically distant populations are one of the most important factors which affects their morphological and physiological variations. These variations are caused by natural mutations accumulated in their habitats. It has been reported that allelic expression differences in the hybrids of genetically distant populations are different from parental strains. In that case, there is a possibility that allelic expression changes lead to novel phenotypes in hybrids. Based on genomic information of the genetically distant populations, quantification and comparison of allelic expression changes make importance of regulatory sequences (cis-acting factors) or upstream regulatory factors (trans-acting modulators) for these changes clearer. In this study, we focused on two Medaka inbred strains, Hd-rR and HNI, derived from genetically distant populations and their hybrids. They are highly polymorphic and we can utilize whole-genome information. To analyze allelic expression changes, we established a method to quantify and compare allele-specific expressions of 11 genes between the parental strains and their reciprocal hybrids. In intestines of reciprocal hybrids, allelic expression was either similar or different in comparison with the parental strains. Total expressions in Hd-rR and HNI were tissue-dependent in the case of HPRT1, with high up-regulation of Hd-rR allele expression in liver. The proportion of genes with differential allelic expression in Medaka hybrids seems to be the same as that in other animals, despite the high SNP rate in the genomes of the two inbred strains. It is suggested that each tissue of the strain difference in trans-acting modulators is more important than polymorphisms in cis-regulatory sequences in producing the allelic expression changes in reciprocal hybrids.     

Genetic relationship between the neurological (excitability thresholds) and the behavioral characteristics of mouse strains and their hybrids

The comparative genetic analysis of 16 inbred mouse strains was carried out for excitability thresholds of peripherical parts of the nervous system, viability to learning, short-term memory and exploratory activity in different experimental conditions. The high positive correlation was shown between excitability thresholds of different parts of the peripherical nervous system, the high negative correlation being established between excitability thresholds, viability to learning, short-term memory and exploratory activity. The data led to the conclusion about the existence of the pleiotropic effect of genes controlling the excitability thresholds, on peculiarities of behaviour. This was confirmed by the findings obtained in the study of various hybrids, which point to the similar way of inheritance of excitability thresholds and behavioural patterns.     

Genotype-temperature interaction inDrosophila melanogaster. I. Viability

Some isogenic strains produced from a natural population at El Prat (Barcelona) were tested at different temperatures for total viability. Their hybrids were also tested at the same temperatures for the same trait. Hybrids proved to be more homcostatic than their isogenic parental strains. More efficient homeostasis in hybrids is suggested to be the cause of increased hybrid superiority with decreasing temperature.Genotype-temperature interaction was always significant for isogenic strains, but never for hybrids. These results are discussed in terms of coadaptation and homeostasis. Some maternal effects have also been detected.     

Chromosome markers in 12 inbred strains of the Norway rat, Rattus norvegicus.

Morphological details of metaphase chromosomes were compared among 12 inbred strains of rats (Rattus norvegicus) by means of conventional Giemsa staining and by a sequential Q- and C-banding method. Inter-strain variations were found in seven pairs, as identified on the basis of size differences in the short arms and/or satellites of chromosomes 3 and 12 and the X chromosome and in the centromeric C-bands of chromosomes 4, 5, 7, and 9. All pairs were homomorphic in the inbred strains, while F1 hybrids between two inbred strains showed certain heteromorphic pairs expected from the parents. These chromosome markers appear to be useful for characterization of inbred strains as well as for various genetic studies, including linkage analyses.     

A major gene affecting age-related hearing loss is common to at least ten inbred strains of mice

Inbred strains of mice offer promising models for understanding the genetic basis of human presbycusis or age-related hearing loss (AHL). We previously mapped a major gene affecting AHL in C57BL/6J mice. Here, we show that the same Chromosome 10 gene (Ahl) is a major contributor to AHL in nine other inbred mouse strains-129P1/ReJ, A/J, BALB/cByJ, BUB/BnJ, C57BR/cdJ, DBA/2J, NOD/LtJ, SKH2/J, and STOCK760. F1 hybrids between each of these inbred strains and the normal-hearing inbred strain CAST/Ei retain good hearing, indicating that inheritance of AHL is recessive. To follow segregation of hearing loss, F1 hybrids were backcrossed to the parental strains with AHL. Auditory-evoked brain-stem response thresholds were used to assess hearing in more than 1500 N2 mice and analyzed as quantitative traits for linkage associations with Chromosome 10 markers. Highly significant linkage was found in all nine strain backcrosses, with the highest probability (LOD > 70) near the marker D10Mit112. This map position for Ahl is near the waltzer mutation (v) and the modifier of deaf waddler locus (mdfw), suggesting the possibility of allelism. Results from an intercross of C57BL/6J and NOD/LtJ mice indicate that the 6- to 10-month difference in AHL onset between these two strains is not due to allelic heterogeneity of the Ahl gene.     

Genetic analysis of susceptibility to isoniazid-induced seizures in mice

Four sets of recombinant inbred lines of mice have been used to analyze genetic differences in acute toxicity of the drug, isonicotinic acid hydrazide. Standard inbred strains, their F₁ hybrids and recombinant inbred strains were all challenged with a single dose of the drug. The percent mortality of the different groups was analyzed to estimate heritability and the number of genes affecting resistance. The data indicated that resistance factors were dominant, heritability was moderate (.25-.37), and more than one gene was involved in each of four different sets of recombinant inbred lines. Possible approaches for identifying and mapping individual genes affecting resistance are discussed.     

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     

Urinary proteins: interstrain differences in the laboratory rat

By comparing electrophoretic patterns of urinary proteins from males of different inbred strains and pertinent hybrids, a new polymorphic locus designated Mur-2 has been detected.     

Genotypic effects on gonadal size in fetal mice

Fetal gonadal size was measured on Days 13, 16 and 19 of gestation in the C57BL/6ByEss (B) and BALB/cByEss (C) inbred strains, their two reciprocal F1 hybrids (CXB and BXC) and in the CXBD and CXBE recombinant inbred lines. At Day 13, CXB F1 fetuses, with C57 fathers and BALB mothers, had significantly larger testes and ovaries than did fetuses of the other 5 stocks. On Day 16, BALB fetuses had significantly larger testes than did C57, while at Day 19 C57 fetuses had significantly larger testes than did BALB fetuses. The CXB and BXC F1 fetuses had significantly larger testes than did mice of the two parental strains on Days 16 and 19, even though the mothers of all 4 kinds of fetus came from the same two inbred strains. C57 and BALB mice did not differ significantly in ovarian size, but had significantly smaller ovaries than did mice of the other genotypes on Days 16 and 19. CXBD mice had the largest ovaries, followed by those of the F1 hybrids. Ovarian size in CXBE mice was similar to that in the CXB hybrids. There were strong maternal effects on gonad size on Days 13 and 19 of gestation. The genes that influenced fetal testicular and ovarian growth appeared to differ from those expressed post-natally at 30 and 60 days.     

Patterns of puffing activity and chromosomal polymorphism in Drosophila subobscura

Summary The effect of inbreeding on the puffing patterns of polytene chromosomes of Drosophila subobscura was analysed. Puffing activity was studied in two strains of D. subobscura: one which had been subject to inbreeding for 288 generations, and in the hybrids from a cross between them. A strong overall decrease in puffing activity was found in the inbred line. In general, hybrids behaved in a similar way to the inbred line or showed activity intermediate between the two lines. The fertility and viability of the two homozygous lines and of the hybrids were also determined. These parameters of fitness are very low in the inbred line. Hybrids displayed intermediate behaviour.Publication No. 109, Departamento de Genética, Universidad de Valencia, Spain     

Behavioural and physiological characterization of inbred mouse strains: prospects for elucidating the molecular mechanisms of mammalian learning and memory

With the advent of recombinant DNA methodology, it has become possible to dissect the molecular mechanisms of complex traits, including brain function and behaviour. The increasing amount of available information on the genomes of mammalian organisms, including our own, has facilitated this research. The present review focuses on a somewhat neglected area of genetics, one that involves the study of inbred mouse strains. It is argued that the use of inbred mice is complementary to transgenic approaches in the analysis of molecular mechanisms of complex traits. Whereas transgenic technology allows one to manipulate a single gene and investigate the in vivo effects of highly specific, artificially induced mutations, the study of inbred mouse strains should shed light on the roles of naturally occurring allelic variants in brain function and behaviour. Systematic characterization of the behavioural, electrophysiological, neurochemical, and neuroanatomical properties of a large number of inbred strains is required to elucidate mechanisms of mammalian brain function and behaviour. In essence, a 'mouse phenome' project is needed, entailing the construction of databases to investigate possible causal relationships amongst the phenotypical characteristics. This review focuses on electrophysiological and behavioural characterization of mouse strains. Nevertheless, it is emphasized that the full potential of the analysis of inbred mouse strains may be attained if techniques of numerous disciplines, including gene expression profiling, biochemical analysis, and quantitative trait loci (QTL) mapping, to name but a few, are also included.     

Relationship between frequencies of univalents and meiotic segregation in different mouse strains and their hybrids

Although univalents in diakinesis are often used as an estimator for chromosome mis-segregation during meiosis, no clear-cut relationship was demonstrated between both phenomena. In this study, the frequencies of autosomal and gonosomal univalents in diakinesis were related to the frequencies of aneuploid metaphase-II gonocytes during spermatogenesis and oogenesis of different mouse strains and their hybrids (inbred strains: DBA/2J, C₅₇Bl; outbred strain; Swiss, inbred x outbred hybrids: Swiss × C₅₇Bl, C₅₇Bl × Swiss, inbred x inbred hybrids: DBA/2J × C₅₇Bl, C₅₇Bl × DBA/2J). As far as the frequencies of univalents are concerned, they were shown to be strain-dependent and similar in both sexes. Moreover, there is a high non-disjunction rate of DBA males and PMSG-HCG-primed DBA females. Aneuploidy in metaphase II is also strain-dependent but different in both sexes; in the male, a clear decrease of aneuploidy frequencies is observed as compared to the frequency of univalents. This decrease does not occur in females.     

Influence of genetic factors on the fertilization of mouse ova in vitro

In-vitro fertilization was studied in F1 hybrids, 4 inbred and outbred strains of mice. Experiments were performed with intact ova (Series I) and denuded ova (Series II). In Series I the highest percentages of fertilization were obtained with gametes of F1 hybrids (86.7%) and of the CBA strain (87.5%). The KP, KE, and particularly the C57 strains, which in vivo give less than 100% fertilization, also gave low indices in vitro (42.9, 26.0 and 8.6% respectively). Testing gametes of these inbred strains with F1 gametes showed that spermatozoa and ova are responsible for the low percentages of fertilization. The rate of fertilization was mainly dependent on the genotype of the spermatozoon. In Series II, high percentages of fertilization (97-100%) were regularly obtained in all groups, indicating that differences between strains pertained mainly to binding of spermatozoa with the zona pellucida. The incidence of polyspermy, which did not exceed 5% in Series I and reached 58% in Series II, was dependent on the genotype of both gametes.     

Calpastatin level in spontaneously hypertensive rats

We studied calpastatin activity in erythrocytes of Milan hypertensive and prehypertensive rats, in their normotensive controls, in F1 and F2 hybrids, and in two inbred strains derived from F2, one hypertensive and the other normotensive. Our results show that the decrease in calpastatin activity observed in Milan hypertensive rats was not caused by hypertension, it was transmitted in a recessive way in heterozygous, and it was not correlated to hypertension.