Jiri Forejt

A Mouse Speciation Gene Encodes a Meiotic Histone H3 Methyltransferase

Speciation genes restrict gene flow between the incipient species and related taxa. Three decades ago, we mapped a mammalian speciation gene, hybrid sterility 1 (Hst1), in the intersubspecific hybrids of house mouse. Here, we identify this gene as Prdm9, encoding a histone H3 lysine 4 trimethyltransferase. We rescued infertility in male hybrids with bacterial artificial chromosomes carrying Prdm9 from a strain with the “fertility” Hst1f allele. Sterile hybrids display down-regulated microrchidia 2B (Morc2b) and fail to compartmentalize γH2AX into the pachynema sex (XY) body. These defects, seen also in Prdm9-null mutants, are rescued by the Prdm9 transgene. Identification of a vertebrate hybrid sterility gene reveals a role for epigenetics in speciation and opens a window to a hybrid sterility gene network.

The European dimension for the mouse genome mutagenesis program

The European Mouse Mutagenesis Consortium is the European initiative contributing to the international effort on functional annotation of the mouse genome. Its objectives are to establish and integrate mutagenesis platforms, gene expression resources, phenotyping units, storage and distribution centers and bioinformatics resources. The combined efforts will accelerate our understanding of gene function and of human health and disease.

Genetic studies on male sterility of hybrids between laboratory and wild mice (mus musculus l.).

The genetic control of the sterility of male hybrids between certain laboratory and wild mice ( Mus musculus L.) is investigated. The observed sterility is, by definition, hybrid sterility since both parental forms (i.e. wild and laboratory mice) are fully fertile, their male offspring displaying small testes with arrest of spermatogenesis at the stage of spermatogenesis or primary spermatocytes. Results of genetic analysis as well as the failure to detect any chromosomal rearrangements point to a genie rather than a chromosomal type of hybrid sterility. Fifty-three wild males were classified into three sets, after mating with C57BL/10 inbred females, according to the fertility of their male progeny (set I – only sterile sons; set II – only fertile sons; set III – both fertile and sterile sons). The wild males of set I, which yield only sterile male offspring with C57BL/10 females, sire sterile sons also with females of the following inbred strains: A/Ph, BALB/c, DBA/1, and AKR/J, whereas the same wild males produce fertile offspring with females of C3H/Di, CBA/J, P/J, I/St and F/St inbred strains. The described hybrid sterility seems to be under the control of several independently segregating genes, one of them (proposed symbol Hst-1) being localized on chromosome 17 (linkage group IX), 6 cM distally from dominant T ( Brachyury ). A chance to search for the mechanism of hybrid sterility is provided by the finding of two laboratory inbred strains, C57BL/10 and C3H/Di, differing with respect to the Hybrid sterility genetic system only at the Hst -1 gene. Hst -1 is closely linked but apparently not identical with the sterility factor of recessive t alleles of the T locus.

Mouse consomic strains: Exploiting genetic divergence between Mus m. musculus and Mus m. domesticus subspecies

Consomic (chromosome substitution) strains (CSs) represent the most recent addition to the mouse genetic resources aimed to genetically analyze complex trait loci (QTLs). In this study, we report the development of a set of 28 mouse intersubspecific CSs. In each CS, we replaced a single chromosome of the C57BL/6J (B6) inbred strain (mostly Mus m. domesticus) with its homolog from the PWD/Ph inbred strain of the Mus m. musculus subspecies. These two progenitor subspecies diverged less than 1 million years ago and accumulated a large number of genetic differences that constitute a rich resource of genetic variation for QTL analyses. Altogether, the 18 consomic, nine subconsomic, and one conplastic strain covered all 19 autosomes, X and Y sex chromosomes, and mitochondrial DNA. Most CSs had significantly lower reproductive fitness compared with the progenitor strains. CSs homosomic for chromosomes 10 and 11, and the C57BL/6J-Chr X males, failed to reproduce and were substituted by less affected subconsomics carrying either a proximal, central, or distal part of the respective chromosome. A genome-wide scan of 965 DNA markers revealed 99.87% purity of the B6 genetic background. Thirty-three nonsynonymous substitutions were uncovered in the protein-coding regions of the mitochondrial DNA of the B6.PWD-mt conplastic strain. A pilot-phenotyping experiment project revealed a high number of variations among B6.PWD consomics.

Mechanistic basis of infertility of mouse intersubspecific hybrids

Significance Hybrid sterility contributes to speciation by restricting gene flow between related taxa. Although four hybrid sterility genes have been identified in Drosophila and mouse so far, the underlying molecular mechanisms are largely unknown. We describe extensive asynapsis of chromosomes in male and female meiosis of F1 hybrids between two closely related mouse subspecies. Using the intersubspecific chromosome-substitution strains, we demonstrate that the heterospecific pairing of homologous chromosomes is a preexisting condition of asynapsis and may represent a universal mechanism of pachytene arrest in interspecific hybrids. Sex-specific manifestation of asynapsis can explain the mechanism of Haldane’s rule. According to the Dobzhansky–Muller model, hybrid sterility is a consequence of the independent evolution of related taxa resulting in incompatible genomic interactions of their hybrids. The model implies that the incompatibilities evolve randomly, unless a particular gene or nongenic sequence diverges much faster than the rest of the genome. Here we propose that asynapsis of heterospecific chromosomes in meiotic prophase provides a recurrently evolving trigger for the meiotic arrest of interspecific F1 hybrids. We observed extensive asynapsis of chromosomes and disturbance of the sex body in >95% of pachynemas of Mus m. musculus × Mus m. domesticus sterile F1 males. Asynapsis was not preceded by a failure of double-strand break induction, and the rate of meiotic crossing over was not affected in synapsed chromosomes. DNA double-strand break repair was delayed or failed in unsynapsed autosomes, and misexpression of chromosome X and chromosome Y genes was detected in single pachynemas and by genome-wide expression profiling. Oocytes of F1 hybrid females showed the same kind of synaptic problems but with the incidence reduced to half. Most of the oocytes with pachytene asynapsis were eliminated before birth. We propose the heterospecific pairing of homologous chromosomes as a preexisting condition of asynapsis in interspecific hybrids. The asynapsis may represent a universal mechanistic basis of F1 hybrid sterility manifested by pachytene arrest. It is tempting to speculate that a fast-evolving subset of the noncoding genomic sequence important for chromosome pairing and synapsis may be the culprit.

Meiotic studies of translocations causing male sterility in the mouse

Unusual meiotic behavior of the XY chromosome pair was observed in sterile male mice doubly heterozygous for two Robertsonian translocations, Rb(16.17)7Bnr and Rb(8.17)1Iem. Nonrandom association between the X chromosome and the translocation configuration, ascertained from the frequencies of relevant C-band contacts, was found in 9 of 10 sterile males. Besides the nonrandom association, the XY chromosomes showed signs of impaired condensation, as judged by measurement of their lengths at diakinesis/MI of the first meiotic division. In contrast, neither nonrandom contact nor decondensation of the XY chromosomes pair was found in fertile males heterozygous for a single Robertsonian translocation, Rb1Iem or Rb7Bnr. The present observations lend indirect support to the working hypothesis advanced previously, the assumption that interference with X-chromosome inactivation is a possible cause of spermatogenic breakdown in carriers of various male-sterile chromosomal transloations. Alternative explanations of the available data, which cannot be ruled out, are briefly discussed.

PPARG by dietary fat interaction influences bone mass in mice and humans

Adult BMD, an important risk factor for fracture, is the result of genetic and environmental interactions. A quantitative trait locus (QTL) for the phenotype of volumetric BMD (vBMD), named Bmd8, was found on mid‐distal chromosome (Chr) 6 in mice. This region is homologous to human Chr 3p25. The B6.C3H‐6T (6T) congenic mouse was previously created to study this QTL. Using block haplotyping of the 6T congenic region, expression analysis in the mouse, and examination of nonsynonymous SNPs, peroxisome proliferator activated receptor γ (Pparg) was determined to be the most likely candidate gene for the Bmd8 QTL of the 630 genes located in the congenic region. Furthermore, in the C3H/HeJ (C3H) strain, which is the donor strain for the 6T congenic, several polymorphisms were found in the Pparg gene. On challenge with a high‐fat diet, we found that the 6T mouse has a lower areal BMD (aBMD) and volume fraction of trabecular bone (BV/TV%) of the distal femur compared with B6 mice. Interactions between SNPs in the PPARG gene and dietary fat for the phenotype of BMD were examined in the Framingham Offspring Cohort. This analysis showed that there was a similar interaction of the PPARG gene and diet (fat intake) on aBMD in both men and women. These findings suggest that dietary fat has a significant influence on BMD that is dependent on the alleles present for the PPARG gene.

Xy pair associates with the synaptonemal complex of autosomal male-sterile translocations in pachytene spermatocytes of the mouse (mus musculus).

Analysis of the chromosome behaviour at pachytene has been performed by means of the silver staining technique visualizing the synaptonemal complexes (SCs) in male mice heterozygous for the male-sterile translocations T(5;12)31H, T(16;17)43H and T(7;19)145H, respectively. The T(9;17)138Ca male heterozygotes and T43H/T43H homozygous males were used as fertile controls. The sterile mice displayed a high frequency (about 60%) of pachytene spermatocytes with autosomal translocation configuration located in close vicinity of the XY pair. The dense round body (XAB), normally located near the X-chromosome axis in fertile males, exhibited abnormal affinity to translocation configuration in the sterile translocation heterozygotes. The incomplete synapsis of autosomes involved in translocation configuration was observed in more than 70% of the pachytene spermatocytes with the male-sterile translocations but in less than 20% of the cells from T138Ca fertile male.s. A hypothesis relating the spermatogenic arrest of carriers of male-sterile rearrangements to the presumed interference with X chromosome inactivation in male meiosis is discussed.

Nonrandom association between a specific autosome and the X chromosome in meiosis of the male mouse: possible consequence of the homologous centromeres’ separation

A nonrandom association is described between chromosome No. 15, which is involved in the T(14;15)6Ca translocation, and the X chromosome in diakinesis/ metaphase I plates of the male mouse. The tight attachment of centromeric heterochromatin (CH) regions of chromosomes X and 15 is predominantly found in those primary spermatocytes in which the distance between the homologous CH regions of the No. 15s exceeds some critical value. The rule holds true for both T6 homozygotes (i.e., for those of their primary spermatocytes displaying two 15t univalents) and T6 heterozygotes (i.e., for translocation configurations “III + I” and certain types of “chain-of-four” quadrivalents). The tight X-15 CH association is highly specific, since out of 1540 primary spermatocytes of the T6 heterozygotes, only 3 showed the tight X-14 CH association. In contrast to the tight CH attachment, the frequency of loose X-15 chromosome association does not show dependence upon the centromere separation of the No. 15s. The nature of the described CH association remains to be clarified; for the present, it seems that the occurrence of a nucleolus organizer on the translocated chromosome 15t presumably plays a role in the causal mechanism of the loose X-15t CH association. Some circumstantial evidence indicates the adverse effect of the observed tight CH attachment on the course of spermatogenesis.

Centromeric Heterochromatin Polymorphism in the House Mouse

Polymorphism of Giemsa-specific centromeric heterochromatin (C.H.) has been described in the laboratory and wild mice. All examined wild mice and inbred mouse strains display some chromosomes with considerably reduced or enlarged C.H. regions. The quantity of C.H. could be an inherent property of a chromosome as inferred from (a) the finding of the identical C.H. pattern within inbred strains, (b) the finding that two genetically related inbred strains, C3H and CBA, separated from each other for more than 150 generations, possess the same two chromosome pairs with tiny C.H. marker regions. These chromosomes were identified as No. 1 (l.g. XIII) and No. 14 (l.g.III) by means of T(14;15)6Ca translocation, and C- and G-band analysis. The neutrality of C.H. polymorphism in murine genome is inferred from the “heterozygosity” for the C.H. variants found in all studied wild mice. The possible relationship of C.H. polymorphism to the centromere interference phenomenon is hypothesized.