6-Sulphatoxymelatonin secretion in different locomotor activity types of the blind mole rat Spalax ehrenbergi.

6-Sulphatoxymelatonin (aMT6S) excretion was examined in the urine of rhythmic and arrhythmic blind subterranean mole rats (Spalax ehrenbergi) to test the correlation between melatonin secretion (as represented by aMT6S) and variability in circadian locomotor activity. Activity pattern was tested in four males, first for a week under short photoperiod [light:dark (LD) 10:14], followed by 10 days in constant darkness (DD). After several months the experiment was repeated under long photoperiod (LD 14:10), followed by DD conditions. Under LD conditions all animals exhibited aMT6S excretion during the dark phase, with a decline just before the onset of light. No correlation was found between activity pattern and melatonin secretion. The animal with the highest melatonin secretion both under LD and DD had an arrhythmic locomotor pattern. The results suggest that in mole rats melatonin secretion and circadian locomotor activity are controlled by two different mechanisms. There were large differences in the aMT6S levels among individuals, suggesting the importance of duration of melatonin secretion over amplitude for gonadal development and thermoregulatory changes. During summer, i.e., before the breeding season, the animals keep a more stable aMT6S secretion than in winter, and the amplitude of secretion is higher under DD vs. LD conditions.

Circadian rhythm and the per ACNGGN repeat in the mole rat, Spalax ehrenbergi.

Individual variability in circadian locomotor activity has recently discovered in the blind mole rat, Spalax ehrenbergi. An interesting association was found between different circadian types and two DNA fragments, 5.6 and 5.9 kb long, that contain the ACNGGN repeat sequence, homologous to a part of the period gene of Drosophila. Nine of 12 arrythmic animals showed the 5.6-kb band, while 13 of 17 circadian rhythmic animals had the 5.9-kb band. This repeat exists also in the brain RNA of the mole rat, apparently in higher quantities during the sleeping phase, suggesting that an unusual protein(s), composed of a poly-Thr-Gly segment, affects in circadian rhythm.

Activity pattern and rhythm in the subterranean mole rat superspecies Spalax ehrenbergi.

Good candidates for naturally occurring variability in circadian rhythms may be subterranean herbivores, since they are not normally subjected to entraining light stimulation. To test this possibility, we selected the blind mole rat Spalax ehrenbergi superspecies in Israel and tested it in short- and long-term experiments. Short-term experiments showed that the animals exhibited three patterns of activity: a regular circadian rhythm (26.6%), an altered circadian rhythm (shorter or longer than normal, 53.1%), and an arrhythmic pattern (20.3%). A long-term experiment showed that the arrhythmic pattern indeed reflected a genuine arrhythmic genotype. The mole rats were found to be active less than 25% of the day and exhibited a multiphasic mode of activity, both diurnally and nocturnally. The number of activity periods and the level of activity were negatively correlated: Animals that exhibited a high level of activity per unit of time showed low numbers of activity periods, while animals that exhibited a lower level of activity showed higher numbers.

The differences among Jewish communities--maternal and paternal contributions.

The haplotypes of Y chromosome (paternally inherited) and mtDNA (maternally inherited) were analyzed in representatives of six Jewish communities (Ashkenazic, North African, Near Eastern, Yemenite, Minor Asian/Balkanian, and Ethiopian). For both elements, the Ethiopian community has a mixture of typically African and typically Caucasian haplotypes and is significantly different from all others. The other communities, whose haplotypes are mostly Caucasian, are more closely related; significant differences that were found among some of them possibly indicate the effects of admixture with neighboring communities of non-Jews. The different contribution of the Y chromosome and mtDNA haplotypes to the significant differences among the communities can be explained by unequal involvement of males and females in the different admixtures. In all communities, except the Ethiopians, the level of diversity (h) for Y chromosome haplotypes is higher than that for mtDNA haplotypes, suggesting that in each community the people who become parents include more males than females. An opposite proportion (more females than males) is found among the Ethiopians.

Mitochondrial DNA affinity of several Jewish communities.

The mitochondrial DNA (mtDNA) of 332 individuals from Israel, including 270 Jews (originating from 7 communities) and 62 Arabs, was analyzed. Each mtDNA haplotype was determined by the fragment patterns of restriction enzymes HpaI, BamHI, HaeII, MspI (HpaII), and AvaII. The variability of the total sample and of each community was high. Of 40 different haplotypes, 20 were found more than once. Most haplotypes are typical of Caucasians, but African types were found among Ethiopian Jews and to a lesser extent among Arabs. The communities differed in their haplotypes: Chi-square tests among six communities showed significant differences for most pairwise comparisons and nonsignificant differences involving mainly the Moroccan Jews. In a genetic distance analysis only the Ethiopian Jews appeared to be distinguished from the other communities. According to a GST analysis, approximately 30% of the variation among the mtDNA restriction maps is attributable to differences between communities.

Origins of H-2 polymorphism in the house mouse. II. Characterization of a model population and evidence for heterozygous advantage.

Comparison of the rate of synonymous and nonsynonymous nucleotide substitutions suggests that certain regions of the functional H-2 genes, which are part of the mouse major histocompatibility complex (Mhc), are under strong positive selection pressure. Thus far, however, little evidence has been provided for the existence of such pressure in natural mouse populations. We have, therefore, initiated experiments designed to test the hypothesis of positive selection acting on H-2 loci. The experiments are being carried out on two natural mouse populations in Jerusalem, Israel. One population occupies a space of about 100 m2 in a chicken coop, the other lives in a nearby field in which "mouse stations" providing food and shelter have been set up. Extensive typing of these two populations revealed the presence of only four H-2 haplotypes. Mice in the two populations breed continually all year around, yet population size varies seasonally, with population maxima in winter and minima in summer. The population in the chicken coop contains a relatively stable nucleus which may be organized in demes with an excess of females over males and limited territorial mobility. The rest of the mice stay in the population for a short time only and then either die or emigrate. The field population is smaller and more loosely organized than the chicken-coop population, with demes probably forming only during population maxima. For the rest of the time breeding in this population is probably panmictic. At a population minimum in the summer of 1984, H-2 homozygotes happened to predominate over heterozygotes. This situation, however, lasted for a short time only and thereafter there was a continuous, statistically highly significant increase in the proportion of H-2 heterozygotes of one or two types. The increase occurred in both populations but was more apparent in the chicken-coop population. This observation provides the first experimental evidence that heterozygous advantage might be one of the mechanisms maintaining high H-2 polymorphism in natural populations of the house mouse.

mtDNA polymorphism in two communities of Jews.

Twenty-one types of mtDNA were found in a survey of 39 Israeli Jews, of whom 18 were Sephardic and 21 Ashkenazic. The survey was made with six restriction enzymes that together recognize an average of 600 bp/genome. The differences among the types appear to be due to base substitution at 19 cleavage sites, one deletion, and one conformational mutation. The numbers of differences imply that these modern Jews stem from a minimum of 21 maternal lineages that were already distinct from one another 4,000-5,000 years ago. In three of the four cases where a type was found in more than one person, it occurred in both Ashkenazic and Sephardic populations. The diversity of types in the combined sample of two Jewish populations is lower than both that in a sample from various parts of Africa and that in a sample from various parts of East Asia. Nevertheless, it is as high as that in a sample from diverse parts of New Guinea, an area much larger than that in Israel to which the Jewish population traces back.

t-Specific DNA polymorphisms among wild mice from Israel and Spain.

Lehrach and his coworkers have isolated a series of DNA probes that specifically hybridize with different regions of mouse chromosome 17 within the t complex. The probes display restriction fragment length polymorphisms, RFLPs, which are specific for the t haplotypes in all laboratory mouse strains tested thus far. Some of these probes have been used to test wild mice populations for these t-associated DNA forms. It is demonstrated that populations from Germany, Switzerland, Italy, Greece, Yugoslavia, Australia, Costa Rica, and Venezuela contain chromosomes in which all the tested DNA loci display the t-specific polymorphisms. The frequency of mice carrying these chromosomes is as high as 31%. Wild mice from Israel and Spain, on the other hand, carry chromosomes displaying t-specific DNA forms only at one or two of the probed loci, while the other loci carry the wild-type (+) forms. These chromosomes thus resemble the partial t haplotypes known from the study of laboratory mice. One possible interpretation of these findings is that these DNA polymorphisms contributed to the assembly of the complete t haplotypes and that these haplotypes may have originated in the Middle East.

Polymorphism of unique noncoding DNA sequences in wild and laboratory mice.

Two DNA probes, D17Tu1 and D17Tu2, were isolated from a genomic DNA library containing only two mouse chromosomes, one of which is chromosome 17, carrying the major histocompatibility complex (H-2), as well as the t complex genes. The D17Tu1 probe was mapped to the centromeric region of chromosome 17 and the D17Tu2 probe to the S region of the H-2 complex. Neither of the two probes appeared to detect any genes, but both contained unique, nonrepetitive sequences. Typing of DNA obtained from a large panel of mice revealed the presence of four D17Tu1 patterns in inbred mouse strains, one very common, one less common, and two present in one strain each. The two common patterns could not be detected in appreciable frequencies in the European wild mice tested (one of the two patterns was, however, found in Australian wild mice). Conversely, the patterns found frequently in European wild mice are absent in the laboratory mice. We therefore conclude that wild mice from the sampled regions of Europe could not have provided the ancestral stocks from which inbred strains were derived. Only one D17Tu1 pattern was found in all the populations of Mus musculus tested, while eight patterns were found in Mus domesticus, with virtually all the populations being polymorphic. We suggest that this difference reflects different modes in which the two species colonized Europe. The distribution of the D17Tu2 patterns in inbred strains correlates with the distribution of H-2 haplotypes.

Low H-2 polymorphism in some Israeli wild mouse populations.

Two populations of the wild house mouse, Mus domesticus, found living close to each other (one inhabited a chicken coop and the other an open field at the Educational Farm of the Hebrew University of Jerusalem, East Talpiot, Jerusalem) were studied for their H-2 polymorphism. These two populations were selected because they are well characterized in terms of their ecological parameters; they have been under continuous surveillance for several years. Twenty-seven H-2 homozygous lines were produced by mating wild mice from these two populations with laboratory strains. The H-2w homozygotes were then characterized by serological typing with monoclonal and polyclonal antibodies specific for the known allomorphs controlled by the class I H-2K and H-2D loci or the class II H-2A and H-2E loci. They were also used as donors for immunizations and for the selection of antisera defining the H-2 haplotypes carried by these lines. Four new H-2 haplotypes could be identified: H-2w82 (Kw16 Dw82), H-2w83 (Kw83 Dw16), H-2w84 (Kw84 Dw84), and H-2w85 (Kw83 Dw84), the last haplotype being a recombinant derived from H-2w83 and H-2w84. Antisera defining the new haplotypes were then used for a study of the wild populations. This study revealed that the populations contain only the four identified H-2 haplotypes, having three alleles at the H-2K locus (Kw16, Kw83, Kw84) and three alleles at the H-2D locus (Dw16, Dw82, and Dw84). The alleles occur in the populations with a frequency of 0.12-0.54. There were no significant differences in gene frequencies between the two populations, and the allele frequencies remained more or less stable. There was a significant excess of heterozygotes for at least some of the genes, compared with the frequency expected from Hardy-Weinberg equilibrium. The same antisera were also used to type other populations in the vicinity of Jerusalem. In one population, located 30 km west of Jerusalem, the mice failed to react with any of the reagents. In the other two populations, located 15 km west and 40 km northeast of Jerusalem, three of the four H-2 haplotypes found in East Talpiot were present at high frequencies. It appears, therefore, that only three main H-2 haplotypes and two or three minor ones are present in the area around Jerusalem. This study thus provides the first example of a large mainland population in which the H-2 polymorphism is comparable to that of many other non-H-2 loci.

Mitochondrial DNA evolution in mice.

This study extends knowledge of mitochondrial DNA (mtDNA) diversity in mice to include 208 animals belonging to eight species in the subgenus Mus. Highly purified mtDNA from each has been subjected to high-resolution restriction mapping with respect to the known sequence of one mouse mtDNA. Variation attributed to base substitutions was encountered at about 200 of the 300 cleavage sites examined, and a length mutation was located in or near the displacement loop. The variability of different functional regions in this genome was as follows, from least to most: ribosomal RNA, transfer RNA, known proteins, displacement loop and unidentified reading frames. --Phylogenetic analysis confirmed the utility of the Sage and Marshall revision of mouse classification, according to which there are at least four species of commensal mice and three species of aboriginal mice in the complex that was formerly considered to be one species. The most thoroughly studied of these species is Mus domesticus, the house mouse of Western Europe and the Mediterranean region, which is the mitochondrial source of all 50 of the laboratory strains examined and of the representatives of wild house mice introduced by Europeans to North and South America during the past few hundred years. --The level of mtDNA variation among wild representatives of M. domesticus is similar to that for the Eastern European house mouse (M. musculus) and several other mammalian species. By contrast, among the many laboratory strains that are known or suspected to stem from the pet mouse trade, there is little interstrain variation, most strains having the "old inbred" type of domesticus mtDNA, whose frequency in the 145 wild mice examined is low, about 0.04. Also notable is the apparent homogeneity of mtDNA in domesticus races that have fixed six or more fused chromosomes and the close relationship of some of these mtDNAs to those of karyotypically normal mice. --In addition, this paper discusses fossil and other evidence for the view that in mice, as in many other mammals, the average rate of point mutational divergence in mtDNA is 2-4% per million years. From this, it is estimated that the commensal association between mice and our ancestors began more than a million years ago, i.e., at an early stage in the evolution of Homo erectus.

Unusual type of mitochondrial DNA in mice lacking a maternally transmitted antigen.

Mice that lack a maternally transmitted antigen (Mta) on the cell surface share a distinctive type of mitochondrial DNA. This is evident from restriction analyses of mitochondrial DNAs from 25 strains of mice whose antigenic state is known. One hundred sixty-eight cleavage sites have been mapped in the mitochondrial DNA of Mta- mice. Detailed maps for the 8 other types of mitochondrial DNA detected in the survey have also been prepared. The Mta- mice are estimated to differ from those expressing the antigen by 108 to 141 base substitutions at widely scattered points in the mitochondrial genome.

Flow of mitochondrial DNA across a species boundary.

Restriction analysis shows that wild Scandinavian mice belonging to the species Mus musculus contain the mitochondrial DNA of a neighboring species, M. domesticus. This demonstration results from comparisons of Scandinavian mice with authentic M. domesticus and M. musculus from other parts of Europe. Electrophoretic and immunological analysis of eight diagnostic proteins confirms that mice from north of the hybrid zone in Denmark are M. musculus in regard to their nuclear genes. In contrast, the mice tested from this region and a nearby part of Sweden have exclusively M. domesticus types of mitochondrial DNA. Phylogenetic analysis of the restriction maps suggests that the mitochondrial DNAs found in Scandinavian M. musculus could stem from a single M. domesticus female.

East Asian hemoglobin type (Hbbp) in wild populations of the house mouse in Israel.

Electrophoretic studies of hundreds of individuals showed that all wild populations of the house mouse in Israel are polymorphic for alleles Hbbd and Hbbp of the hemoglobin locus. No mouse carrying Hbbs was found. This finding contradicts the notion that Hbbp is limited to East Asian house mice.

The genetic basis of egg lay response to conditioned medium in the flour beetle, Tribolium castaneum : I. Two-way selection.

The rate of egg laying in Tribolium castaneum is affected by the quality of the environment and can thus serve not only as a component of fitness of the individual, but also as an adaptation to the long-term survival of the population. In an attempt to determine whether it is an independent genetic character, selection for high and low rates of response was carried jut in beetles from three wild populations. When tests were done on virgin females, and responsiveness, corrected for scale effects, was used as the criterion for selection, separation between the lines was achieved in one generation. It is suggested that in natural populations of Tribolium, the responsiveness of the rate of egg laying to the quality of the environment is controlled by a small number of genes.