Heteromorphic X chromosomes in 46,XX males: Evidence for the involvement of X-Y interchange

Summary G- and R-banded chromosome preparations from eight of twelve 46,XX males, with no evidence of mosaicism or a free Y chromosome, were distinguished in blind trials from preparations from normal 46,XX females by virtue of heteromorphism of the short arm of one X chromosome. Photographic measurements on X chromosomes and on chromosome pair 7 in cells from twelve 46,XX males, eight 46,XX females, and four 46,XY males revealed a significant increase in the size of the p arm of one X chromosome in the group of XX males, independently characterised as being heteromorphic for Xp. No such differences were observed between X chromosomes of normal males and females or between homologues of chromosome pair 7 in all groups. The heteromorphism in XX males is a consequence of an alteration in shape (banding profile) and length of the tip of the short arm of one X chromosome, and the difference in size of the two Xp arms in these 46,XXp+ males ranged from 0.4% to 22.9%. From various considerations, including the demonstration of a Y-specific DNA fragment in DNA digests from nuclei of one of three XX males tested, it is concluded that the Xp+ chromosome is a product of Xp-Yp exchange. These exchanges are assumed to originate at meiosis in the male parent and may involve an exchange of different amounts of material. The consequences of such unequal exchange are considered in terms of the inheritance of genes located on Yp and distal Xp. No obvious phenotypic difference was associated with the presence or absence of Xp+. Thus, some males diagnosed as 46,XX are mosaic for a cryptic Y-containing cell line, and there is now excellent evidence that maleness in others may be a consequence of an autosomal recessive gene. The present data imply that in around 70% of 46,XX males, maleness is a consequence of the inheritance of a paternal X-Y interchange product.     

X-Y exchanges in males of Drosophila hydei carrying the w m Co duplication

Males carrying, inserted on their Y chromosome, a small fragment of X including the w ⁺ (and N ⁺) locus (white-mottled Confluens, w ^m Co), were crossed with the purpose of scoring exceptional progeny. Some of the male and female exceptions were progeny tested and further analysed. Among the various mechanisms which may lead to exceptional offspring, X-Y exchanges proved to occur with a not negligible frequency. The rate was 3%. Nondisjunction accounts for the bulk of the remaining exceptions and appears to be increased considerably in the presence of rearrangements on one or the other of the sex chromosomes.The w ^m Co fragment after having been switched from Y to X by some mechanism other than regular crossing over, may become retransferred to a normal Y chromosome, but at a rate below 3%.     

Chiasmata and chromosome breakages are related to crossing over in Drosophila ananassae males.

A cytogenetic analysis of male crossing over in Drosophila ananassae revealed that cytological exchanges resulted in genetic crossing over, and that chiasma frequency and the genetic recombination correlated positively in chromosomes 2 and 3. Furthermore, the frequency of chromosome breakages correlated positively with chiasma frequency. Paracentric inversion heterozygosity had no detectable influence on the chromosome pairing or exchange events within the inversion loop at meiosis. Scoring of the chiasma demonstrated that males homozygous for the previously mapped enhancers of male crossing over had low frequencies of chiasmata, whereas higher frequencies of chiasmata were observed in males heterozygous for enhancers. The results presented here indicate that the genetic factors controlling male crossing over are involved in the origin of chromosome breakages and in exchange events.     

Accidental X-Y recombination and the aetiology of XX males and true hermaphrodites

Accidental recombination between the differential segments of the X and Y chromosomes in man occasionally allows transfer of Y-linked sequences to the X chromosome leading to testis differentiation in so-called XX males. Loss of the same sequences by X-Y interchange allows female differentiation in a small proportion of individuals with XY gonadal dysgenesis. A candidate gene responsible for primary sex determination has recently been cloned from within this part of the Y chromosome by Page and his colleagues. The observation that a homologue of this gene is present on the short arm of the X chromosome and is subject to X-inactivation, raises the intriguing possibility that sex determination in man is a quantitative trait. Males have two active doses of the gonad determining gene, and females have one dose. This hypothesis has been tested in a series of XX males, XY females and XX true hermaphrodites by using a genomic probe, CMPXY1, obtained by probing a Y-specific DNA library with synthetic oligonucleotides based on the predicted amino-acid sequence of the sex-determining protein. The findings in most cases are consistent with the hypothesis of homologous gonad-determining genes, GDX and GDY, carried by the X and Y chromosomes respectively. It is postulated that in sporadic or familial XX true hermaphrodites one of the GDX loci escapes X-inactivation because of mutation or chromosomal rearrangement, resulting in mosaicism for testis and ovary-determining cell lines in somatic cells. Y-negative XX males belong to the same clinical spectrum as XX true hermaphrodites, and gonadal dysgenesis in some XY females may be due to sporadic or familial mutations of GDX.     

Properties of the sex chromosomes of Lucilia cuprina deduced from radiation studies

From a study of radiation-induced X-chromosome deletions the locus of black body (b) has been localized to the proximal portion of C-band defined euchromatin. Radiation produced mostly X-chromosome deletions rather than point mutations, total X or Y chromosome loss through breakage, or increased frequency of non-disjunction. Aberrant sex ratios obtained indicate that the X chromosome carries vital loci that were deleted with b ⁺ in many cases. The X/O karyotype produces fertile adult females with a characteristic phenotype which is also produced by X deletions. Sex chromosome non-disjunction to give X/O females and X/X/Y males is normally rare but is enhanced by the presence of chromosome rearrangements even when the X and Y are not involved.     

Further studies on the interchromosomal effect on crossing over in Drosophila melanogaster affecting the preconditions of exchange

The second chromosome inversion In (2L+2R) Cy in a heterozygous condition was studied for its effect on frequency and interference of crossing over in three different regions of the X chromosome of Drosophila melanogaster. A significant increase in crossing over frequency was observed in the proximal and distal regioins of the X chromosome while in the middle of the chromosome crossing over frequency remained unaltered. The effect on interference remained unaltered at both ends of the X chromosome while a significant decrease was observed in the middle of the chromosome. These results suggest that the interchromosomal effect on crossing over affects the preconditions of exchange differently in different regions of the X chromosome, and possibly the duration of chromosome pairing.     

Induced crossing-over in the presterile broods of drosophila melanogaster males

The results from two experiments on induced crossing-over in the germ cells of adultDrosophila melanogaster males, heterozygous for 12–13 third-chromosome mutant loci, support the premise that the F₁ offspring recovered in the 6–8 day broods are derived mainly from cells that had been primary spermatocytes at the time of irradiation. This conclusion is also rather well supported by the results from similar experiments reported in the literature. The frequency of exchange in the third-chromosome following irradiation of the adult male is in the order of 0.01% for the 1–3 and 4–5 day broods, 0.15% for the 6–7 day broods and 0.31% for the 8th day brood in the pooled data from 18 experiments. The spontaneous rate for the third-chromosome is in the order of 0.001%. The rather great variance among the experiments in time of recovery of the first crossovers and the frequency of exchange during the time of sampling the spermatocytes and definitive spermatogonia—ranging from none to a 20-fold increase (over the spontaneous rate) in the 6–8 day broods, and from none to a 10–50-fold increase from the 6–8 to the 9–11 day broods — can be attributed to the brood procedures or mating pressure, the effectiveness of ascertainment of crossing-over, particularly in the early broods, and the dosages used by the different investigators. The lack of a dosefrequency relationship in the broods prior to the period of peak sterility is due, at least in part, to lethal-loss of the cells with induced exchange. The evidence from the two experiments also supports the premise that there may be different types of exchange-induction with a causal relationship between the time of induction and the site in the chromosome. The higher relative frequency of multiple crossovers and noncentric exchanges among the recombinants recovered in the 1–7 day broods, as compared to their frequency in the spermatogonial broods, suggests that there may be a difference in the mode of induction or transmission of recombinant chromosomes in different stages of spermatogenesis. The relatively high incidence of F₁-sterility of the recombinants and homozygous lethality of the recombinant chromosomes points to the possibility that either the meiotically-induced exchanges are intrachromosomal aberrations frequently accompanied by duplications or deficiencies due to breakage at non-identical loci, or homologous chromosomes sensitive to the induction of exchange are more likely to be hypersensitive to the induction of genetic damage with detrimental fertility and viability effects.     

Chromosomal aberration and sister-chromatid exchange frequencies in peripheral blood lymphocytes of a large human population sample

In order to assess the potential of cytogenetic determinations on peripheral blood lymphocytes as a means of monitoring human populations subject to low level occupational and environmental exposures to chemical mutagens and carcinogens, accurate baseline data are required. Accordingly, we have determined mean frequencies of chromosomal aberrations and of sister-chromatid exchanges, their variances, and the sources of this variance in a cohort of 353 healthy employees of the Brookhaven National Laboratory. A detailed protocol was adopted for blood sampling, lymphocyte culture, cytogenetic preparation and scoring in order to minimize variation from these potential sources. Scoring was divided between the Oak Ridge and the Brookhaven groups with duplicate scoring sufficient to evaluate and minimize the effect of any differences between laboratories or between individual scorers. In all, the data include 71,950 cells scored for chromosomal aberrations and 16,898 cells scored for sister-chromatid exchanges. The mean unadjusted frequency of sister-chromatid exchanges was 8.29 +/- 0.08/cell. As reported in other studies, cigarette smoking very significantly influenced sister-chromatid exchange frequencies; in our study the mean for smokers was 9.0 +/- 0.2, while that for non-smokers was 8.1 +/- 0.1/cell. The mean frequency was statistically higher in females than in males, regardless of smoking status. On the other hand, age of the subject did not significantly influence sister-chromatid exchange frequencies. Curiously, the subject's total white cell count did influence sister-chromatid exchange frequency. No other source of variation was found. The frequencies of chromosomal aberrations of all types were determined. The frequency of the most common unequivocal chromatid type, the chromatid deletion, was 0.81 +/- 0.05%, that of the most common unequivocal chromosome type, the dicentric, was 0.16 +/- 0.02%. No statistically significant influence was found of age or sex, nor of any other parameter tested, on the frequency of any chromosomal aberration type, with the single exception of long acentric fragments, often "supernumerary", believed to represent X chromosomes precociously separated at the centromere. Such fragments were significantly more frequent in samples from females than those from males, and showed a significant positive regression on age.     

Adding the heterochromatic YL arm to an X chromosome reduces reproductive fitnesses in Drosophila melanogaster: implications for the evolution of rDNA, heterochromatin, and reproductive isolation

For X-Y exchange to be of importance in the coevolution of X and Y rDNA, there must be a mechanism to maintain cytologically normal X chromosomes in the face of continual infusions of X.YL chromosomes produced by X-Y exchanges. Replicated populations were founded with different frequencies of isogenic X and X.YL chromosomes. The X.YL chromosome declined in frequency over time in all lines. Relative fitnesses, estimated from chromosome frequency trajectories, were 0.40, 1.01, and 1.0 for X.YL/X.YL, X.YL/X, and X/X females and 0.75 and 1.0 for X.YL/Y and X/Y males, respectively. The equilibrium frequency for the X.YL chromosome due to the balance between X-Y exchange and selection was predicted to be 4-16 x 10(-4). The results strengthen the evidence for the involvement of X-Y exchange in the coevolution of X and Y rDNA arrays. Conditions for the evolution of reproductive isolation by sex-chromosome translocation are much less probable than previously supposed since the X.YL translocation chromosome is at a selective disadvantage to cytologically normal X chromosomes. Additional heterochromatin was not neutral but was only deleterious beyond a threshold, as one dose of the heterochromatic XL arm did not reduce female reproductive fitness, but two doses did.     

Steroid sulfatase gene in XX males.

The human X and Y chromosomes pair and recombine at their distal short arms during male meiosis. Recent studies indicate that the majority of XX males arise as a result of an aberrant exchange between X and Y chromosomes such that the testis-determining factor gene (TDF) is transferred from a Y chromatid to an X chromatid. It has been shown that X-specific loci such as that coding for the red cell surface antigen, Xg, are sometimes lost from the X chromosome in this aberrant exchange. The steroid sulfatase functional gene (STS) maps to the distal short arm of the X chromosome proximal to XG. We have asked whether STS is affected in the aberrant X-Y interchange leading to XX males. DNA extracted from fibroblasts of seven XX males known to contain Y-specific sequences in their genomic DNA was tested for dosage of the STS gene by using a specific genomic probe. Densitometry of the autoradiograms showed that these XX males have two copies of the STS gene, suggesting that the breakpoint on the X chromosome in the aberrant X-Y interchange is distal to STS. To obtain more definitive evidence, cell hybrids were derived from the fusion of mouse cells, deficient in hypoxanthine phosphoribosyltransferase, and fibroblasts of the seven XX males. The X chromosomes in these patients could be distinguished from each other when one of three X-linked restriction-fragment-length polymorphisms was used. Hybrid clones retaining a human X chromosome containing Y-specific sequences in the absence of the normal X chromosome could be identified in six of the seven cases of XX males.(ABSTRACT TRUNCATED AT 250 WORDS)     

Modes of inheritance of X-Y dissociation in inter-subspecies hybrids between BALB/c mice and Mus musculus molossinus

Genetic analysis of the high frequency of X-Y chromosome dissociation found in primary spermatocytes of F1 hybrids between Japanese wild mice (Mus musculus molossinus) and inbred laboratory mice (BALB/c) was attempted. The frequency of X-Y dissociation (X//Y) in both BALB/c and M. m. molossinus was lower than 30% (Low X//Y), while the value was more than 70% (High X//Y) in their F1 hybrids. Two types of progeny (High X//Y and Low X//Y) appeared in the backcross between BALB/c and High X//Y males, although the frequency of Low X//Y progeny decreased with increasing numbers of backcross generations (26.5% at N2, 13.2% at N3, 5.3% at N4, and 0% at N5). Low X//Y sires produced only Low X//Y mice. We hypothesize that at least one heritable factor which is responsible for the end-to-end association of the sex chromosomes (temporally symbolized as Sxa) is located on the common part of the X and Y chromosomes. The Sxa allele of BALB/c is Sxaa and that of M. m. molossinus is Sxab. The genotype expected in High X//Y males is Sxaa/Sxab and in Low X//Y males and their parental stocks either Sxaa/Sxaa or Sxab/Sxab. The repeated segregation of Low X//Y progeny from High X//Y sires is interpreted simply by assuming that crossing-over has occurred between the X and Y chromosomes. The gradual decrease in the recombinant type mice (Low X//Y) during sequential backcrosses suggests the presence of some autosomal factors that suppress the crossing-over of the sex chromosomes and that do not seem to function in the inter-subspecies hybrids.     

Sex chromosomal polymorphism in the earwig Forficula

A four-year population sampling programme revealed small annual and marked seasonal variations in the frequency of adult XXY males of the earwig Forficula auricularia, where XY and XXY individuals co-exist in the same population as a polymorphism. It is suggested that this may have been due to an effect of the extra X chromosome on development rate. The extra X was also found to reduce autosomal chiasma frequency slightly and to change bodily morphometrics: antenna length was increased and head and abdomen length decreased in a compensating fashion. Chiasma and multivalent frequencies were analysed in 50 spontaneously autotetraploid cells. The range of sex chromosome numbers found in different species, and the variation in Forficula due to allosomal polymorphism and non-disjunction, suggests that sex determination in earwigs does not involve a balance mechanism. The role of polyploidy in Dermapteran evolution is supported.Dedicated to Dr. Sally Hughes-Schrader for her many valuable contributions to the field of cytology.     

Cytological studies of meiotic recombination in human males

We combined immunostaining and fluorescence in situ hybridization (FISH) methodology to directly examine meiotic exchanges in over 2,000 pachytene stage spermatocytes from 25 individuals. Our results indicate that, on average, there are about 50 exchanges per cell and that, with the exception of the acrocentric chromosomes, all chromosome arms harbor at least one exchange. We also identified significant among-individual variation in the mean number of exchanges, with an approximate 20% difference between individuals with "low" and those with "high" exchange frequencies.     

Systematic comparison of bacterial feeding strains for increased yield of Caenorhabditis elegans males by RNA interference-induced non-disjunction

Rare Caenorhabditis elegans males arise when sex chromosome non-disjunction occurs during meiosis in self-fertilizing hermaphrodites. Non-disjunction is a relatively rare event, and males are typically observed at a frequency of less than one in five hundred wild-type animals. Males are required for genetic crosses and phenotypic analysis, yet current methods to generate large numbers of males can be cumbersome. Here, we identify RNAi reagents (dsRNA-expressing bacteria) with improved effectiveness for eliciting males. Specifically, we used RNAi to systematically reduce the expression of over two hundred genes with meiotic chromosome segregation functions, and we identified a set of RNAi reagents that robustly and reproducibly elicited male progeny.     

Analysis of two 47,XXX males reveals X-Y interchange and maternal or paternal nondisjunction

Summary Two cases of 47,XXX males were studied, one of which has been published previously (Bigozzi et al. 1980). Analysis of X-linked restriction fragment length polymorphisms revealed that in this case, one X chromosome was of paternal and two were of maternal origin, whereas in the other case, two X chromosomes were of paternal and one of maternal origin. Southern blot analysis with Y-specific DNA probes demonstrated the presence of Y short arm sequences in both XXX males. In one case, the results obtained pointed to a paracentric inversion on Yp of the patient's father. In situ hybridization indicated that the Y-specific DNA sequences were localized on Xp22.3 in one of the three X chromosomes in both cases. The presence of Y DNA had no effect on random X inactivation. It is concluded that both XXX males originate from aberrant X-Y interchange during paternal meiosis, with coincident nondisjunction of the X chromosome during maternal meiosis in case 1, and during paternal meiosis II in case 2.     

The influence of the Robertsonian translocation Rb(X.2)2Ad on anaphase I non-disjunction in male laboratory mice

A Robertsonian translocation in the mouse between the X chromosome and chromosome 2 is described. The male and female carriers of the Rb(X.2)2Ad were fertile. A homozygous/hemizygous line was maintained. The influence of the X-autosomal Robertsonian translocation on anaphase I non-disjunction in male mice was studied by chromosome counts in cells at metaphase II of meiosis and by assessment of aneuploid progeny. The results conclusively show that the inclusion of Rb2Ad in the male genome induces non-disjunction at the first meoitic division. In second metaphase cells the frequency of sex-chromosomal aneuploidy was 10.8%, and secondary spermatocytes containing two or no sex chromosome were equally frequent. The Rb2Ad males sired 3.9% sex-chromosome aneuploid progeny. The difference in aneuploidy frequencies in the germ cells and among the progeny suggests that the viability of XO and XXY individuals is reduced. The pairing configurations of chromosomes 2, Rb2Ad and Y were studied during meiotic prophase by light and electron microscopy. Trivalent pairing was seen in all well spread nuclei. Complete pairing of the acrocentric autosome 2 with the corresponding segment of the Rb2Ad chromosome was only seen in 3.2% of the cells analysed in the electron microscope. The pairing between the X and Y chromosome in the Rb2Ad males corresponded to that in males with normal karyotype. Reasons for sex-chromosomal non-disjunction despite the normal pairing pattern between the sex chromosomes may be seen in the terminal chiasma location coupled with the asynchronous separation of the sex chromosomes and the autosomes.(ABSTRACT TRUNCATED AT 250 WORDS)     

The etiology of maleness in XX men

Summary Information relating to the etiology of human XX males is reviewed. The lesser body height and smaller tooth size in comparison with control males and first-degree male relatives could imply that the patients never had any Y chromosome. Neither reports of occasional mitoses with a Y chromosome, nor of the occurrence of Y chromatin in Sertoli cells are convincing enough to support the idea that low-grade or circumscribed mosaicism is a common etiologic factor. Reports of an increase in length of one of the X chromosomes in XX males are few and some are conflicting. Nor is there any evidence to support the idea of loss of material. However, absence of visible cytogenetic alteration does not rule out the possibility of translocations, exchanges or deletions.A few familial cases are known. Mendelian gene mutations may account for a number of instances of XX males, similar genes being well known in several animal species. The existing geographical differences in the prevalence of human XX males could be explained by differences in gene frequency. But if gene mutation were a common cause of XX maleness there would be more familial cases.Any hypothesis explaining the etiology of XX males should take into account the following facts. There are at least 4 examples of XX males who have inherited the Xg allele carried by their fathers, and at least 9 of such males who have not. The frequency of the Xg phenotype among XX males is far closer to that of males than to that of females, while the absence of any color-blind XX males (among 40 tested) resembles the distribution in females. Furthermore, H-Y antigen is present in XX males, often at a strength intermediate between that in normal males and females. Finally, in a pedigree comprising three independently ascertained XX males, the mothers of all three are H-Y antigen-positive, and the pattern of inheritance of the antigen in two of them precludes X-chromosomal transmission.Many of the data are consistent with the hypothesis that XX males arise through interchange of the testic-determining gene on the Y chromosome and a portion of the X chromosome containing the Xg gene. However, actual evidence in favor of this hypothesis is still lacking, and the H-Y antigen data are not easy to explain. In contrast, if recent hypotheses on the mechanisms controlling the expression of H-Y antigen are confirmed, a gene exerting negative control on testis determination would be located near the end of of the short arm of the X chromosome. This putative gene is believed not to be inactivated in normal females, for at least two other genes located in the same region, i.e. Xg and steroid sulfatase, are not. Deletion or inactivation of these loci would explain how XX males arise and would be consistent with most, but not all, the facts.There is yet no single hypothesis that by itself can explain all the facts accumulated about XX males. While mosaicism appears very unlikely in most cases, Mendelian gene mutation, translocation, X-Y interchange, a minute deletion or preferential inactivation of an X chromosome, or part thereof, remain possible. The etiology of XX maleness may well be heterogeneous.     

Patterns of exchange induced by mitomycin C in C-bands of human chromosomes. I. Relationship to C-band size in chromosomes 1, 9, and 16

Summary Frequencies of exchange were determined in C-bands of chromosomes 1, 9 and 16 in six normal males, and related to relative C-band area. Comparing these different chromosomes, more exchanges occurred on average in 9 than in 1 although their mean C-band sizes were similar. Chromosome 16 exchanges were fewer, both overall and relative to C-band area. Comparing the same chromosome between individuals, there was a positive correlation between relative frequency and band size in both 1-1 and 9-9 exchanges. No clear trend was observed for other exchange events.If homology is required for interchange, if cannot be dependent solely on overall C-band size. Perhaps certain DNA sequences, sensitive to mitomycin C damage, are located in part of each C-band, with less per unit area in chromosome 1 than in 9 and still less in chromosome 16.X- and U-type exchanges between chromosome 9s occurred in near equal frequencies in all individuals. If synapsis of specific, affected sequences is a pre-requisite for interchange, this observation suggests that the affected sequence in chromosome 9 is arranged in both orientations relative to the centromere.     

Meiotic disjunction and embryonic lethality in sex-linked double-translocation heterozygous males of the onion fly,Hylemya antiqua (Meigen)

Summary The frequencies of disjunction types in double-translocation heterozygous males (2⁶2^Y6²6XY²) in Hylemya antiqua have been established in MII cells and eggs of testcrosses.Several disjunction types occurred but four predominated. A correlation was found between the frequencies of the disjunction types and the relative position of the centromeres. The frequency of numerical non-disjunction (NND) was 4%. Differences in frequency of NND between sex-linked and autosomal translocations of H. antiqua are discussed. A good correspondence between the frequencies of unbalanced karyotypes, and embryonic and larval mortality was found. The total genetic load which can be induced by the T14/T61 males is estimated to be 60–65%. Some duplication/deficiency karyotypes appeared to be viable in pupal and even adult stages. In 2⁶2⁶2^Y6²6²X males a regular coorientation between 2^Y and X was observed, in spite of non-homologous centromeres and a complicated synapsis of 2^Y. Application possibilities of the present material for genetic control of H. antiqua are discussed.     

Simultaneous measure of X-ray induced X-Y and autosomal crossingover in spermatocytes ofDrosophila

Summary Radiation induced X-Y and autosomal crossingover was followed in primary spermatocytes ofDrosophila males. Males of the compositiony/B ^Sy⁺ were X-rayed 1–2 hours after the prepupal stage and mated daily with threey;dp cn bw females. The results suggest that the frequency of recovered X-Y crossovers is highest in the late spermatocyte stage about to undergo the first meiotic division, whereas the peak frequency of autosomal crossovers occurs in earlier spermatocytes. The results are explainable if (1) a more appropriate pairing configuration exists between X and Y than between autosomes at later stages of the primary spermatocyte, a situation which is reversed in earlier stages; (2) breakability of Y (and/or X) heterochromatin and centromeric heterochromatin of autosomes are different from each other at different stages of the primary spermatocyte; or (3) a combination of both.Supported by National Science Foundation grant GB 3117.