This Special Issue of Genetical Research constitutes a tribute to the work and achievements of Bruce Cattanach on the occasion of his official retirement. Bruce began his career working on mutagenesis in mice. His interest in mutagenesis continued throughout his career but through it he was led to a wide range of major discoveries in mammalian genetics. The contributions from his colleagues in this issue provide a flavour of the various fields his work has covered.

Quantitative trait loci (QTLs) responsible for variation in sternopleural bristle number in crosses between the laboratory lines of Drosophila melanogaster OregonR and CantonS were mapped using information from allele frequency changes of two families of retrotransposon markers in divergently selected populations. QTL effects and positions were inferred by likelihood, using transition matrix iteration and Monte Carlo interval mapping. Individuals from the selected populations were genotyped for markers spaced at an average distance 4.4 cM. Four QTLs of moderate effect ranging from 0·6 to 1·32 bristles accounted for most of the selection response. A permutation test of the correspondence between the mapped QTLs and the positions of bristle number candidate genes suggested that alleles at these candidate genes were no more strongly associated with selected changes in marker allele frequency than were randomly chosen positions in the genome.

We present data on the relationship between the rate of transposition and copy number in the genome for the copia and Doc retrotransposons of Drosophila melanogaster. copia and Doc transposition rates were directly measured in sublines of the isogenic 2b line using individual males or females, respectively, with a range of copia copy numbers from 49 to 103 and Doc copy numbers from 112 to 235 per genome. Transposition rates varied from 3×10−4 to 2×10−2 for copia and from 2×10−4 to 2×10−3 for Doc. A positive relationship between transposition rate and copy number was found both for copia and for Doc when the data were analysed across all the 2b individuals; no significant correlation was found when the data were analysed across the subline means for both retrotransposons tested. Overall, correlation between copia and Doc transposition rate and their copy number in the genome, if any, was not negative, which would be expected if transposable elements (TEs) self-regulate their copy number. Thus, for copia and Doc no evidence for self-regulation was provided, and at least for these two TEs this hypothesis is not favoured for explaining the maintenance of the stable copy number that is characteristic for natural populations. The transposition rate of copia was measured twice, and a strong positive correlation between copy number and transposition rate both across individuals and subline means was found in 1994, while in 1995 no correlation was found. This fact is in agreement with the hypothesis that a positive correlation between the rate of transposition and TE copy number may be a default starting point for future host–TE coevolution.

To test whether stressful conditions altered levels of heritable variation in fecundity in Drosophila melanogaster, parent–offspring comparisons were undertaken across three generations for flies reared in a combined stress (ethanol, cold shock, low nutrition) environment or a control environment. The stressful conditions did not directly influence fecundity but did lead to a reduced fecundity in the offspring generations, perhaps reflecting cross-generation maternal effects. Both the heritability and evolvability estimates were higher in the combined stress treatment, reflecting an apparent increase in the additive genetic variance under stress. In contrast, there were no consistent changes in the environmental variance across environments.

Ifosfamide induced dominant lethal mutations in spermatozoa of mice at doses of 200 and 300 mg/kg and in spermatids and spermatocytes at 600 mg/kg. The highest dose also induced specific-locus mutations in post-spermatogonial germ-cell stages of mice but not in spermatogonial stem cells. The nature of the induced mutations suggests they are intergenic. The spermatogenic specificity of ifosfamide in mouse germ cells is similar to that of the structurally related cytostatic drugs cyclophosphamide and trofosfamide. Due to the post-spermatogonial germ cell specificity of ifosfamide, the genetic risk is limited to a few weeks after exposure.

Based on the outbred mouse strain Fzt: Du, which has been obtained by systematic crossing of four inbred and four outbred lines, a long-term selection experiment was carried out for total protein amount (PA) in the carcass, starting in 1975. An unselected control line (CO) was kept under the same management but without continuous protein analysis. The protein amount of male carcasses at 42 days of age (P42) increased from 2·9 g in generation 0 to 5·2 g at generation 70, representing 97% of a theoretical selection limit. The total selection response amounts to 2·3 g, which is about 80% above the initial value and corresponds to 9σp or 12σA . The estimated realized heritability of protein amount decreased from 0·56 to 0·03 at generation 70, which was due to an increase in phenotypic variance from 0·065 to 0·24 g2 and a reduction in genetic variance from 0·04 to 0·01 g2. Half the selection response was obtained after about 18 to 23 generations, a half-life of 0·25 to 0·3 Ne. The maximum selection response was 0·094 g/generation and the response was 0·01 g/generation at generation 70. The measurements of body weights at 0, 10, 21, 42 and 63 days throughout the experiment showed a strong correlated effect for all weights. The PA mice are one of the heaviest lines of mice ever reported, and do not differ significantly in their body composition from control mice at 42 days. The direct selection response was due primarily to increased general growth. Body weight and protein amount are phenotypically and genetically highly correlated (rp=0·82, rA≈1); however, selection for body weight led to fatter animals, whereas selection for protein opposed increased fatness (at least until selection age). This may be of general importance in animal breeding. The comparatively high selection response in this experiment seems due to the heterogeneity of the base population, the relatively high effective population size, and the duration of the experiment.

The radioprotective effects of misoprostol, a synthetic stable analogue of prostaglandin E1, on spermatogonial stem cells of C3H/HeH×101/F1 hybrid mice (3H1) were analysed by establishing dose–response relationships for stem cell killing by X-rays in mice that were pretreated with misoprostol. Spermatogonial stem cell killing was studied through determination of the percentage of tubular cross-sections showing repopulation at 10 days after irradiation. In control mice, the D0 values ranged between 1·7 and 3·6 Gy, dependent on the stage of the cycle of the seminiferous epithelium the cells were in. As found previously, proliferating spermatogonial stem cells were much more radioresistant than quiescent stem cells. In the misoprostol-pretreated animals the spermatogonial stem cells were more radioresistant, the D0 values ranging from 3·6 to 5·0 Gy. Both proliferating and quiescent spermatogonial stem cells were protected by misoprostol. As the dose–response curves in control and misoprostol-pretreated mice showed about the same extrapolation number to the y-axis it was concluded that the misoprostol pretreatment did not alter the kinetics of the repopulation process.

Ritter & Salamini (1996) presented a systematic account of two-point linkage analysis in allogamous diploid plant species. Vowden et al. (1995) described an alternative approach that is implemented in a computer program LINKEM. This paper describes how the latter approach has been extended to three-point linkage analysis, and implemented in a new program LINK3EM that is available from the authors. The essence of the approach is for the computer program to derive the appropriate form of analysis for a specific cross from its ‘knowledge’ of the most general type of cross that can arise. This avoids the need for programming specific codes for the many different types of cross that can arise. The program allows different locus orderings and parental phases to be compared. The Haldane or Kosambi map functions can be specified, although it is also possible to estimate all three pairwise recombination fractions without any assumed map function.

A set of Drosophila melanogaster was generated, all derived from a common isogenic base stock and each with a single new P element insert on the second or third chromosome. The lines were scored for their body size, measured as thorax length. P inserts were associated with highly significant effects on body size, although the genotypes of the construct and of the control prevented deduction of the direction of mutant effects. In addition to mutant effects on the thorax length of both sexes, there were also highly significant sex-specific effects. Pleiotropic effects of inserts affecting body size on viability and bristle number, as ascertained in a separate study of these lines (Lyman et al., 1996), were weak. Insertional mutagenesis is potentially a powerful tool for investigating the genes involved in size-control in Drosophila, but the technique requires fine tuning for use on polygenic and fitness-related traits.

When a favourable mutation sweeps to fixation, those genes initially linked to it increase in frequency; on average, this reduces diversity in the surrounding region of the genome. In the first analysis of this ‘hitch-hiking’ effect, Maynard-Smith and Haigh (1974) followed the increase of the neutral allele that chanced to be associated with the new mutation in the first generation, and assumed that the subsequent increase was deterministic. Later analyses, based on either coalescence arguments, or on diffusion equations for the mean and variance of allele frequency, have also made one or both of these assumptions. In the early generations, stochastic fluctuations in the frequency of the selected allele, and coalescence of neutral lineages, can be accounted for correctly by following relationships between genes conditional on the number of copies of the favourable allele. This analysis shows that the hitch-hiking effect is increased because an allele that is destined to fix tends to increase more rapidly than exponentially. However, the identity generated by the selective sweep has the same form as in previous work, h[r/s] (2 Ns)−2r/s, where h[r/s] tends to 1 with tight linkage. This analysis is extended to samples of many genes; then, genes may trace back to several families of lineages, each related through a common ancestor early in the selective sweep. Simulations show that the number and sizes of these families can (in principle) be used to make separate estimates of r/s and Ns.

The effects of cancer predisposition and increased tumorigenic radiosensitivity of the predisposed genotypes on radiation cancer risks (in the general population and in sisters and first cousins of affected probands) are studied using an autosomal dominant model of cancer predisposition and radiosensitivity. The model assumes that the predisposing alleles, which confer enhanced tumorigenic radiosensitivity, are incompletely penetrant. In addition, the model also allows for sporadic cancers, unrelated to the predisposing locus. The predictions of the model are illustrated using current estimates of BRCA1 mutant gene frequencies; the estimates of the strength of predisposition and radiosensitivity differentials used are based on animal and human studies. It is shown that, unless both the strength of predisposition and radiosensitivity differential are large (say, >100-fold in comparison with normal homozygotes), (i) the effect of risk heterogeneity on cancer risk is marginal; (ii) dose-dependent radiation effect remains virtually the same as in a homogeneous irradiated population that has no predisposed subgroups; (iii) for the same radiation dose, relatives of affected probands show an enhancement of cancer risks; and (iv) most extra cancers in relatives can be attributed to radiosensitivity differentials. This simple model can give an upper bound of the effect of risk heterogeneity on radiation-induced breast cancer risks even when the cumulative breast cancer risk is age-dependent. Further, our model predicts that the benefits of mammography outweigh the risks.

We investigated three aspects of adaptation to variable environments in Daphnia pulex (Cladocera: Crustacea): (1) effects of temporal variation on the evolution of phenotypic plasticity ; (2) plasticity in sexual versus asexual lineages; (3) maintenance of genetic variation in variable environments. We performed a 72-day quasi-natural selection experiment comparing three patterns of variation: constant temperatures, varying but predictable temperature change, and unpredictable temperature change. All populations were begun with an identical array of 34 clones. During selection clonal variation declined in all populations and different patterns of environmental variation had little effect on amounts of genetic variation. Sexual and asexual lineages differed in size and growth rate, but did not differ in amounts of plasticity or in adaptation to variable environments. The primary target of selection was the Malthusian parameter (r) and life history traits of development time, offspring size and offspring number. The heritability of plasticity was generally lower than trait heritability. Because of this difference, the selection response on the mean of the traits overwhelmed the selection response on plasticity. Lower heritabilities of plasticity are very typical, suggesting that our results will be typical of responses to selection in nature. Our results suggest that selection will act mostly on trait means within environments and that plasticity will evolve often as a correlated trait. Because selection on plasticity is based on its across-deme, global fitness, this process will usually be slow. Comparative studies need to shift from closely related, local population differences to those of more distantly related populations or even different species.

Temporally varying selection is considered to be one of the potential mechanisms of recombination evolution. We found earlier that simple cyclical selection for a trait controlled by multiple additive, dominant or semi-dominant loci can result in extremely complex limiting behaviour (CLB) of population trajectories, including ‘supercycles’ and more complex attractors. Recombination rate proved to be a key factor affecting the mode of CLB and the very existence of CLB. Therefore, we considered here a generalized model: the fixed recombination rate was replaced by a polymorphic recombination modifier. The modifier-dependent changes included: (a) supercyclical dynamics due to the recombination modifier in a system that does not manifest CLB when recombination rate is a fixed parameter; (b) appearance of a new level of superoscillations (super-supercycles) in a system that manifests supercycles with a fixed modifier; (c) chaotization of the regular supercyclical dynamics. The domain of attraction of these movements appeared to be quite large. It is noteworthy that the modifier locus is an active participant in the observed non-monotonic limiting movements. Interactions between short-period forced oscillations and the revealed long-period auto-oscillations appeared to result in new regimes of recombination evolution (for some range of linkage between the modifier locus and the selected system), as compared with those caused by the forced oscillations alone.

A study was undertaken to test whether the elimination of metabolic pathways strongly involved in growth and fatness, comprising thyroid hormones (TH) and growth hormone (GH), is responsible for a substantial part of the genetic change produced by selection. Lines used in this study have been selected for about 50 generations for high (PH) and low (PL) body weight at 10 weeks and for high (F) and low fat content (L) at 14 weeks, producing a 3-fold difference in body weights and a 5-fold difference in fat content. Thyroid ablation was achieved by repeated backcrossing into the four selection lines of a transgene comprising the HSV1-tk gene coupled to the promoter of the thyroglobulin gene. Hemizygous pregnant dams were treated with ganciclovir leading to thyroid-ablated dams and offspring and therefore to a lack of TH and subsequently of GH. In the absence of TH and GH, lines still differ in body weight over the period studied (10 d to about 100 d; e.g. at the end PH=32·1 g vs PL=10·2 g) and in fat content (F=16·2% vs L=3·8%) ; the corresponding values for the wild-type controls were PH=49·9 g vs PL=17·4 g and F=27·5% vs L=4·8%. The effect of the transgene depended on the genetic background for body weights at most ages and for relative gonadal fat pad weights, but less for fat content. The L line showed the lowest growth depression. The lit gene, which causes GH but not TH deficiency, was also transferred by repeated backcrosses into three of these lines (PH, PL, F). The combined deficiency of TH and GH had bigger effects on body weights at earlier ages than did GH deprivation. The data show that changes in the TH- and GH-systems are not the only cause of line differences in growth and fatness resulting from long-term selection, but both are involved to a significant extent. The interactions between the effects of the transgene and of the lit gene and the genetic background were, nevertheless, relatively small and therefore these results support a polygenic model of selection response.

The occurrence of homozygous-viable dilute–short ear (Myo5a–Bmp5) double mutants in mouse specific locus mutation experiments has generally been assumed to be the result of double non- disjunction such that the mutant inherits two copies of chromosome 9 carrying the recessive alleles from the test-stock. A homozygous viable Myo5a–Bmp5 double mutant was recovered recently in our laboratory. We were able to genetically analyse both the Myo5a–Bmp5 region and proximal and distal markers in the original mutant as well as in offspring of the original mutant. Our results indicate the mutational event to be due to mitotic recombination and not double non-disjunction.

The patch (Ph) locus allele, patch-extended (Phe), has significantly less pigmentation than the original mutation and homozygotes have been known to survive to term. Analysing inter- subspecific F1 hybrids, we were able to demonstrate that Phe is a deletional mutation encompassing the platelet-derived growth factor receptor alpha subunit (Pdgfra). The deletion does not appear to extend into the coding sequence of the Kit gene (a related tyrosine kinase receptor). However, we were able to demonstrate that, while the Kit gene is transcribed, it does not encode a functionally active receptor.

Formulae are given for computing the distribution of numbers of selected individuals of each genotype and thus change in gene frequency at a locus with a large effect on a quantitative trait under truncation selection in a finite population. Results are illustrated with respect to use of selection for quantitative trait locus (QTL) detection, specifically by bulk segregant analysis with linked markers, for which probabilities that selected samples will comprise almost all one genotypic class are computed.

Methods of identification of quantitative trait loci (QTL) using a half-sib design are generally based on least-squares or maximum likelihood approaches. These methods differ in the genetical model considered and in the information used. Despite these differences, the power of the two methods in a daughter design is very similar. Using an analogy with a one-way analysis of variance, we propose an equation connecting the two test-statistics (F ratio for regression and likelihood ratio test in the case of the maximum likelihood). The robustness of this relationship is tested by simulation for different single QTL models. In general, the correspondence between the two statistics is good under both the null hypothesis and the alternative hypothesis of a single QTL segregating. Practical implications are discussed with particular emphasis on the theoretical distribution of the likelihood ratio test.

We report here a novel X-linked mutant, named faint lined (Fnl), which was discovered in the litter of an irradiated 3H1 male (Dr Bruce Cattanach, personal communication). The mutation is associated with fine dorsal striping in affected heterozygous females and prenatal lethality in males. Approximately 50% of Fnl/+ females die in utero and surviving animals have a reduced weight at birth and weaning. Histological studies failed to reveal the underlying basis of the phenotype or any gross structural abnormalities in internal organs (Fnl/+×Mus spretus) F1 affected females were backcrossed to 3H1 males and haplotype analysis positioned Fnl in the proximal region of the mouse X chromosome distal to Ant2 and proximal to Hprt. Therefore, Fnl lies within a defined conserved segment and its human homologue can be predicted to lie in the ANT2–HPRT region in Xq25. Further genetic resolution of co-segregating markers flanking Fnl established that Fnl lies in a 7·6±2·6 cM interval between DXMit50 and DXMit82.