Jacob Wahrman

Meiotic association between the XY chromosomes and unpaired autosomal elements as a cause of human male sterility

Intimate association between autosomal translocation trivalents and XY bivalents at pachytene was observed in a majority of cells of two men ascertained through primary sterility and found to be heterozygous for a 14;21 Robertsonian translocation. The association, studied by light and electron microscopy of spread first spermatocytes, was between the unpaired short arms of the normal chromosomes of the translocation trivalent and the differential axes of the XY chromosomes. In a minority of cells, this contact was not established, or not maintained, as alternative combinations between the elements available for non-homologous pairing were realized. Following a suggestion of Lifschytz and Lindsley (1972), sterility in these patients was attributed to spermatogenic arrest caused by physical contact of sex chromosomes with autosomal material and consequent interference with the normal metabolism of the sex chromosomes. Autosomal aberrations and polymorphisms, which lead to the presence of unpaired segments at meiosis, may thus play a critical role in a general mechanism of chromosomally-derived male sterility. It is proposed that such a mechanism may also be instrumental in the initiation of reproductive barriers in nature.

Xist RNA is associated with the transcriptionally inactive XY body in mammalian male meiosis

Abstract.In eutherian mammals, X inactive-specific transcripts (Xist) are expressed in somatic cells possessing more than one X chromosome, and in germline cells of males, in which the single X chromosome is transcriptionally inactive. In early meiosis of males the sex chromosomes form an inactive XY nuclear compartment (XY body). We show by in situ reverse-transcribed polymerase chain reaction that Xist RNA is concentrated in the XY body. This fine localization suggests that Xist RNA is involved in inactivation of the male X chromosome, and that it has spreading capability, not only in cis but also in a quasi-cis mode, to juxtaposed non-X chromosomes. A hypothetical scheme links the evolution of heteromorphic sex chromosomes to the development of X condensation/inactivation in the male. The mechanism of X inactivation in somatic cells of mammalian females, resulting in male/female dosage compensation, has been recruited from the Xist-activated chromosome condensation machinery that developed in male meiosis earlier in evolution.

Histone macroH2A1.2 is concentrated in the XY compartment of mammalian male meiotic nuclei

We show here that histone macroH2A1.2 concentrates at the transcriptionally silent XY body, normally being formed during male meiosis in the mouse. A similar accumulation has earlier been observed on the inactive X chromosomes of somatic adult female mammalian cells by Costanzi and Pehrson (1998). This correspondence in the nature of heterochromatinization of the X chromosomes in males and females adds another property of X chromosome inactivation that is shared by males and females at different phases of their life cycle.

Modified testicular expression of stress-associated "readthrough" acetylcholinesterase predicts male infertility

Male infertility is often attributed to stress. However, the protein or proteins that mediate stress‐related infertility are not yet known. Overexpression of the “readthrough” variant of acetylcholinesterase (AChE‐R) is involved in the cellular stress response in a variety of mammalian tissues. Here, we report testicular overexpression of AChE‐R in heads, but not tails, of postmeiotic spermatozoa from mice subjected to a transient psychological stress compared with age‐matched control mice. Transgenic mice overexpressing AChE‐R displayed reduced sperm counts, decreased seminal gland weight, and impaired sperm motility compared with age‐matched nontransgenic controls. AChE‐R was prominent in meiotic phase spermatocytes and in tails, but not heads, of testicular spermatozoa from AChE‐R transgenic mice. Head‐localized AChE‐R was characteristic of human sperm from fertile donors. In contrast, sperm head AChER staining was conspicuously reduced in samples from human couples for whom the cause of infertility could not be determined, similar to the pattern found in transgenic mice. These findings indicate AChE‐R involvement in impaired sperm quality, which suggests that it is a molecular marker for stress‐related infertility.

REVISITING SPALAX: MITOTIC AND MEIOTIC CHROMOSOME VARIABILITY

ABSTRACT Further cytological studies on Israeli mole rats (Spalax) were motivated by their postulated active speciation. Four major chromosomal forms were characterized by Wahrman and collaborators in 1969. The differences between their chromosome numbers, 2n = 52, 54, 58 and 60, were then attributed to 1–4 Robertsonian changes, and the differences in the number of chromosome arms were assumed to be due to pericentric inversions. The new results obtained by differential staining techniques agree with the earlier interpretations. C-banding has shown that some of the evolutionary changes were accompanied by changes in the quantity and distribution of constitutive heterochromatin. All chromosomal forms also possess a considerable amount of chromosome micro-changes, including variation in the length of a C-negative, heterochromatic modification, at the base of the long arm of Chromosome 1. Four chromosomes may carry nucleolus-organizing regions (NORs), one of them in a distal position. Each of five individual...

Mammalian meiosis involves DNA double-strand breaks with 3′ overhangs

Meiotic recombination in yeast is initiated at DNA double-strand breaks (DSBs), processed into 3′ single-strand overhangs that are active in homology search, repair and formation of recombinant molecules. Are 3′ overhangs recombination intermediaries in mouse germ cells too? To answer this question we developed a novel approach based on the properties of the Klenow enzyme. We carried out two different, successive in situ Klenow enzyme-based reactions on sectioned preparations of testicular tubules. Signals showing 3′ overhangs were observed during wild-type mouse spermatogenesis, but not in Spo11−/− males, which lack meiotic DSBs. In Atm−/− mice, abundant positively stained spermatocytes were present, indicating an accumulation of non-repaired DSBs, suggesting the involvement of ATM in repair of meiotic DSBs. Thus the processing of DSBs into 3′ overhangs is common to meiotic cells in mammals and yeast, and probably in all eukaryotes.

The origin of multiple sex chromosomes in the gerbil Gerbillus gerbillus (Rodentia: Gerbillinae)

The sex chromosomes of the partly sympatric species of gerbils Gerbillus pyramidum and G. gerbillus (Mammalia: Gerbillinae) were investigated by a variety of light- and electron-microscope methods, including DNA replication banding and synaptonemal complex (SC) techniques. The sex-chromosome mechanism of G. pyramidum is of the maleXY:femaleXX type, whereas that of G. gerbillus is of the less common maleXY1Y2:femaleXX system. The results include the demonstration that the X chromosomes of both species are compound. One segment is added to the X chromosome of G. pyramidum, leading to an increase in length from the standard 5% to approximately 7.3%, whereas two different extra segments increase the length of the X chromosome of G. gerbillus to approximately 11% of the length of the haploid genome. In both cases the extra material is autosomal and is also represented in the respective Y chromosomes. Classifying heterochromatin by the variation in staining quality was helpful in elucidating the possible origin of the different chromosome segments, including the pericentromeric regions. Observations on meiotic chromosome pairing and chiasma formation have confirmed the homologies established by band comparisons. The occurrence of chiasmata between the sex chromosomes supports the autosomal origin of the pairing segments. These and other findings have been interpreted in the framework of a multistep evolutionary model. This sequence starts from a hypothetical pair of sex chromosomes, the X element of which amounts to 5% of the haploid genome, and leads through three translocations involving two pairs of autosomes and one pericentric inversion to the most complex situation of this series, manifested in G. gerbillus. The adaptive value, if any, of autosome incorporation into the sex chromosomes repeatedly occurring here is unknown. It is, however, a remarkable fact that in one species, G. gerbillus, the complex sex-chromosome constitution is conserved over vast geographic distances, and in the other, G. pyramidum, the compound X and Y chromosomes withstand change in the face of extreme autosome restructuring.

Solitary and synaptonemal complex-associated recombination nodules in pro-nurse cells during oogenesis in Drosophila melanogaster

An oocyte in Drosophila melanogaster originates from 1 of 16 cells comprising an ovarial syncytium. The two pro-oocytes proceed into the pachytene stage of meiosis, but only one develops further into a mature oocyte while the other reverts to a nurse cell. It is known that pro-nurse cells also enter meiosis, as they contain incomplete synaptonemal complexes (SCs). We now show that these cells also harbour recombination nodules (RNs). In cells that only occasionally contain SC segments, the RNs are typically not located close to distinct tripartite SC structures. Instead, these RNs are frequently associated with a spherical body of amorphous material and two to three, more or less parallel fibres, possibly representing SC material. The significance of the solitary RNs is discussed in relation to the present knowledge of the assembly and disassembly of the SC.

Regions of active chromatin conformation in 'inactive' male meiotic sex chromosomes of the mouse.

The sensitivity to DNase I of the meiotic sex chromosomes of the male mouse was determined by in situ nick translation. At pachytene and diakinesis-metaphase I, six segments, four at the ends of the X and Y chromosomes and two at internal sites on the X chromosome, were found to be more sensitive than the other parts of these chromosomes. The sensitive segments presumably reflect an active or potentially active chromatin conformation which is maintained in the sex chromosomes despite the earlier reported, almost complete cessation of uridine incorporation. The distribution of regions which are sensitive to DNase I corresponds to that of early DNA replication bands. Active conformation patterns like those figured here, probably exist in the sex chromosomes of other mammals as well.

Chromosomally derived sterile mice have a ‘fertile’ active XY chromatin conformation but no XY body

We have previously shown that the sex chromosome bivalent of normal, fertile male mice possesses extensive regions of potentially active chromatin, even though, as has been shown by others, certain X-linked genes, and perhaps most of the X chromosome, become inactivated during pachytene. The male meiosis of a fertile (2; 11) translocation carrier mouse, a chromosomally derived sterile (11; 19) translocation carrier and that of normal mice is compared. In situ nick translation shows a similar DNase I sensitivity pattern in the sex chromosomes of all examined mice. The X chromosome has four regions of potentially active chromatin conformation, two at the ends of the chromosome and two interstitial ones, coinciding with flexures which become prominent towards late pachytene. The Y chromosome is almost uniformly sensitive to DNase I. The similarity of chromatin conformation patterns in fertile and sterile mice is compatible with the hypothesis that unscheduled transcription of particular genes, possibly included in the active conformation regions, occurs in mice which become sterile. In the sterile (11; 19) translocation carrier, a vast majority of all pachytenes are “associated”: usually one unpaired segment of chromosome 19 is in end-to-end contact with the X chromosome. The tips of both unpaired segments of chromosome 19 have a thickened axis and display a peculiar chromatin appearance, similar to the modification of the centromeric tip of the X chromosome. Telomeric unpairedness of certain chromosome segments seems to be conducive to ausotome-X chromosome association. We suggest that compartmentalization of the nucleus into an autosome mass and a fully developed, protruding, metabolically quiescent XY body, is a precondition for the normal progressing of meiosis. In the associated cells, the autosomal quadrivalent anchors the XY bivalent among the autosomes; as a consequence no XY body is formed. This interference with the course of compartmentalization leads to the abolishment of inactivation of part or all of the potentially active genes and results in meiotic arrest, and hence in sterility.