Unequal nondisjunction frequencies of trivalent chromosomes in male mice heterozygous for two Robertsonian translocations

Robertsonian (Rb) translocation heterozygosity may cause pairing problems during prophase and segregation irregularities at anaphase of meiosis I. These stages of meiosis I were studied in male mice doubly heterozygous for the two Rb chromosomes Rb(9.19)163H and Rb(16.17)8Lub. At pachytene both Rb chromosomes similarly showed pairing irregularities like unpaired segments. However, highly different nondisjunction frequencies of chromosomes forming the respective trivalents were found. The nondisjunction frequency of the Rb8Lub trivalent chromosomes was about 40%, whereas a very low frequency of nondisjunction was found in combination with the Rb163H trivalent. Since both trivalents were together in the same cell, differences in kinetochore function are assumed to be responsible for the diverse frequency of nondisjunction.     

Immunocytogenetics. III. Analysis of trivalent and multivalent configurations in mouse pachytene spermatocytes by human autoantibodies to synaptonemal complexes and kinetochores

Mice heterozygous for one or more Robertsonian (Rb) translocation chromosomes have been used to analyze synaptonemal complex (SC) configurations and kinetochore arrangements in trivalents and multivalents. Rb heterozygosity without arm homologies leads to the formation of heteromorphic trivalents in meiosis I; alternating homology of the chromosome arms produces ringlike or chainlike multivalents. Immunofluorescence double-labeling with human antibodies to SCs and kinetochores was performed on surface-spread pachytene spermatocytes. Both Rb bivalents and Rb trivalents clearly showed that metacentrics possess only one centromere. In heteromorphic trivalent SCs, the nonhomologous kinetochores of the two acrocentrics were closely paired in a cis-configuration and juxtaposed opposite the kinetochore of the metacentric; the latter appeared to be an integral part of the longitudinal SC axis. Meiotic multivalents of interpopulation hybrids included up to 36 chromosome arms. In multivalent SCs, the kinetochores always lay together, with the SC arms arranged away from the central centromere cluster. The paracentromeric regions of the Rb chromosomes appeared to remain unsynapsed on both sides of the centromeres. The SC arms were often linked by end-to-end associations. Following desynapsis of the multivalent SC, the kinetochores of the Rb metacentrics showed a highly nonrandom topologic distribution within the nucleus, reminiscent of their arrangement during synapsis.     

Synaptonemal complexes and chromosome chains in the rodent Ellobius talpinus heterozygous for ten Robertsonian translocations

Synaptonemal complexes (SC) in four Ellobius talpinus males heterozygous for ten Robertsonian translocations were examined with an electron microscope using a surface-spreading technique. A total of 136 late zygotene and pachytene spermatocytes were examined. From one to three completely paired SC trivalents were found in each early pachytene spermatocyte. The lateral elements of the short arms of the acrocentric chromosomes in these trivalents were joined with an SC thus forming the third arm of the SC trivalent. At the same stage a few SC trivalents did not contain lateral elements in the pericentromeric region of the metacentric chromosomes and remained unpaired in this region up to mid pachytene. At zygotene and pachytene from two to eight SC trivalents were joined into chains due to formation of SCs between the short arms of acrocentrics of other SC trivalents. These chains are frequent at late zygotene, but are resolved during pachytene into individual trivalents. It is proposed that pairing and SC formation between the short arms of the acrocentric chromosomes results from the monosomy of the short arms and partial DNA homology between these heterochromatic regions. Since crossing over probably does not take place in these segments, the chromosomal chains may subsequently be corrected into trivalents by a dissolution of the SCs combining adjacent trivalents. The correction and disjoining of chains may not be effective in all cells. The cells in which the chains are retained are assumed to be arrested at the pachytene stage.     

Trivalent behavior during prophase I in male mice heterozygous for three Robertsonian translocations: an electron-microscopic study

A synaptonemal complex (SC) analysis was carried out in male mice heterozygous (CHT/+) for three Robertsonian translocations. All pachytene preparations studied showed the presence of three trivalents. At early pachytene, the nonhomologous centromeric regions of the acrocentric chromosomes were unpaired. Heterosynapsis subsequently took place with complete pairing of the trivalents. Association between one of the three trivalents and the sex vesicle was observed in 30.4% of the nuclei. Association between the unpaired regions of two trivalents was present in 14.4% of the cells, suggesting that the relationship between unpaired regions of structural rearrangements and the X-Y bivalent may simply reflect the tendency of unpaired regions to establish end-to-end associations or heterosynapses among them, which are usually resolved during the pachytene stage of prophase I. Since the sex bivalent always has unpaired regions, these associations often affect the sex chromosomes.     

Cosegregation of Robertsonian metacentric chromosomes in the first meiotic division of multiple heterozygous male mice as revealed by FISH analysis of spermatocyte II metaphases

Contrasting results (random segregation or cosegregation of isomorphic chromosomes) have been reported up to now on the segregation pattern of Robertsonian metacentric chromosomes of Mus musculus domesticus in multiple heterozygotes, using different approaches (karyotypical analysis of the progeny or of second meiotic metaphases). In the present contribution data are presented based on FISH (Fluorescence In Situ Hybridisation) analysis with telomeric probes, which allowed us to distinguish metacentric chromosomes from pairs of acrocentric chromosomes with their centromeric regions close to each other. Probes were hybridized to DAPI stained metaphases of spermatocytes II of mice heterozygous for two, three or four Robertsonian metacentrics in an all-acrocentric background, the karyotype of which has been reconstructed starting from laboratory strains. Isomorphic chromosomes tend to cosegregate (metacentrics with metacentrics, acrocentrics with acrocentrics); the values found for cosegregation have a clear even if moderate effect on the reproductive isolation caused by underdominant chromosomal rearrangements.     

Synapsis in Robertsonian heterozygotes and homozygotes of Dichroplus pratensis (Melanoplinae, Acrididae) and its relationship with chiasma patterns

Dichroplus pratensis has a complex system of Robertsonian rearrangements with central-marginal distribution; marginal populations are standard telocentric. Standard bivalents show a proximal-distal chiasma pattern in both sexes. In Robertsonian individuals a redistribution of chiasmata occurs: proximal chiasmata are suppressed in fusion trivalents and bivalents which usually display a single distal chiasma per chromosome arm. In this paper we studied the synaptic patterns of homologous chromosomes at prophase I of different Robertsonian status in order to find a mechanistic explanation for the observed phenomenon of redistribution of chiasmata. Synaptonemal complexes of males with different karyotypes were analysed by transmission electron microscopy in surface-spread preparations. The study of zygotene and early pachytene nuclei revealed that in the former, pericentromeric regions are the last to synapse in Robertsonian trivalents and bivalents and normally remain asynaptic at pachytene in the case of trivalents, but complete pairing in bivalents. Telocentric (standard) bivalents usually show complete synapsis at pachytene, but different degrees of interstitial asynapsis during zygotene, suggesting that synapsis starts in opposite (centromeric and distal) ends. The sequential nature of synapsis in the three types of configuration is directly related to their patterns of chiasma localisation at diplotene-metaphase I, and strongly supports our previous idea that Rb fusions instantly produce a redistribution of chiasmata towards chromosome ends by reducing the early pairing regions (which pair first, remain paired longer and thus would have a higher probability of forming chiasmata) from four to two (independently of the heterozygous or homozygous status of the fusion). Pericentromeric regions would pair the last, thus chiasma formation is strongly reduced in these areas contrary to what occurs in telocentric bivalents.     

Synaptonemal complex analysis of heteromorphic trivalents in Lemur hybrids

Synaptonemal complexes (SCs) in surface spread pachytene spermatocytes of Lemur resemble those in other mammals and are of two types: metacentric (or submetacentric) and acrocentric, with a very short second arm. In autosomal SC and mitotic karyotypes of Lemur fulvus (2n=60) a 11 proportionality in relative length is observed as in other mammals. In an intraspecific lemur hybrid (2n=55) obtained by mating L. fulvus rufus (2n=60) x L. fulvus collaris (2n=51), G-band patterns show that 10 single acrocentric mitotic chromosomes correspond to the arms of 5 single metacentrics, implying homology. It is inferred that the metacentrics have evolved by centric (Robertsonian) fusion of the acrocentrics. In the SC karyotype of the hybrid all SCs are normal except for five which have the configurations expected of metacentric-acrocentric trivalents. Similarly, in L. f. collaris (2n= 51), with one unpaired metacentric and two unpaired acrocentrics, one such SC trivalent is present in the complement. In an SC trivalent, each of the acrocentric long axes is synapsed with an arm of the metacentric axis, confirming the homology predicted from banding similarities. At late zygotene, the acrocentric short arms, which are non-homologous, are the last to pair, demonstrating that synapsis of the homologous arms occurs first. At later pachytene the acrocentric short arms are fully synapsed, producing a short SC side arm. This subsequent non-homologous synapsis is taken to be an instance of the synaptic adjustment phenomenon which has been shown to lead to non-homologous synapsis in a duplication and several inversions in the mouse. The kinetochore of the metacentric is the same size as those of the acrocentrics, and thus is unlikely to have arisen by true centromeric fusion, but rather by a translocation. The kinetochores of the acrocentrics always lie together on the same side of the metacentric kinetochore (cis configuration), implying a single pairing face on the metacentric axis. The observed trivalent configuration may well constitute a prerequisite for proper meiotic disjunction in metacentric-acrocentric heterozygotes. Such a mechanism is consistent with fertility regularly observed in such hybrid lemurs.     

Nondisjunction rates of mouse specific chromosomes involved in heterozygous Rb rearrangements measured by chromosome painting of spermatocytes II. I. The effects of the number of trivalents

Dual-colour FISH painting with alternative fluorescent chromosome-specific probes allowed us to distinguish chromosomes 1, 4, 6 and 14. The purpose was to check whether nondisjunction rates of specific chromosomes involved in heterozygous Robertsonian fusions are independent of the number of trivalents, or an epistatic effect among Rb chromosomes takes place affecting nondisjunction rates. Probes were used on DAPI-stained metaphases of spermatocytes II of laboratory strains of mice with reconstructed karyotypes heterozygous for one, two, three or four Robertsonian metacentrics in an all-acrocentric background. The existence of such epistatic interactions was not verified.     

Cytogenetic control of a hybrid zone of between two Sorex araneus chromosome races before breeding season

Two chromosome races of common shrew, Moscow and Seliger, differ in the arm combination in 11 diagnostic chromosomes (Robertsonian metacentrics/acrocentrics). Homozygotes of both pure races, simple Robertsonian heterozygotes of Seliger race, and complex heterozygotes (FI hybrids) were detected in the found earlier between hybrid zone of these races, in the spring before the breeding seasonbreeding season. The g/oheterozygote was first discovered in race Seliger, whose chromosome formula typically contains acrocentrics g and o. The m/q heterozygote was recorded for the second time. Meiosis was studied in 16 males representing five detected karyotypic categories. No abnormal in pairing of homologs in either sex trivalent common for the species (XY1Y2) or autosome trivalents (g/o and m/q) was detected at diakinesis--metaphase I. Two hybrids displayed a theoretically expected and unimpaired meiotic configuration in a form of a very long chain comprising 11 monobrachial homologs (g/gm/mq/qp/pr/rk/ki/ih/hn/no/o). The results are discussed in terms of hypotheses on fertility of complex heterozygotes and limited gene flow in hybrid zone.     

Cytogenetic control of a hybrid zone between two <Emphasis Type="Italic">Sorex araneus</Emphasis> chromosome races before breeding season

Two chromosome races of common shrew, Moscow and Seliger, differ in the arm combination in 11 diagnostic chromosomes (Robertsonian metacentrics/acrocentrics). Homozygotes of both pure races, simple Robertsonian heterozygotes of Seliger race, and complex heterozygotes (F₁ hybrids) were detected in the found earlier hybrid zone of these races, in the spring before the breeding season. The g/o heterozygote was first discovered in race Seliger, whose chromosome formula typically contains acrocentrics g and o. The m/q heterozygote was recorded for the second time. Meiosis was studied in 16 males representing five detected karyotypic categories. No abnormal in pairing of homologs in either sex trivalent common for the species (XY1Y2) or autosome trivalents (g/o and m/q) was detected at diakinesis-metaphase I. Two hybrids displayed a theoretically expected and unimpaired meiotic configuration in a form of a very long chain comprising 11 monobrachial homologs (g/gm/mq/qp/pr/rk/ki/ih/hn/no/o). The results are discussed in terms of hypotheses on fertility of complex heterozygotes and limited gene flow in hybrid zone.     

Synapsis and chiasma distribution in mice heterozygous for translocations in chromosomes 16 and 17

Electron microscopic analysis of synaptonemal complexes and analysis of chiasmata distribution in male mice heterozygous for Robertsonian translocation T(16; 17)7Bnr - (Rb7), for synaptonemal reciprocal translocation T(16;17)43H - (T43), in double heterozygotes for these translocations and in males with partial trisomy of the proximal region of chromosome 17 was carried out. Synaptic disturbances around the breakpoints of the translocations, such as asynapsis of homologous regions of partners and non-homologous synapsis of centromeric regions of acrocentric chromosomes, were revealed. Synaptic regularity in the proximal part of the chromosome 17 appeared to be affected by no t12 haplotype. Good coincidence between sizes of mitotic chromosomes and corresponding lateral elements of synaptonemal complexes was found for all chromosomes, with the exception of Rb7 in trisomics. In the latter karyotype, the proximal part of chromosome 17 involved in Robertsonian fusion seems to be shortened in the course of zygotene and never synapted with homologous segment of neither the acrocentric chromosome 17 nor large product of reciprocal translocation. Drastic increase in chiasmata frequency in the proximal part of chromosome 17 was revealed in heterozygotes for T43H and in trisomics, as compared with the double heterozygotes Rb7/T43. The latter finding was explained by the existence of two independent pairing segments in the former karyotypes.     

Pachytene pairing and oocyte numbers in mice with two single Robertsonian translocations and the male-sterile compound with monobrachial homology

Oocyte numbers and synaptonemal complexes were studied in two Robertsonian translocations, Rb(6.15)1Ald and Rb(4.6)2Bnr, and their male-sterile compound. Oocyte numbers in the compound were lower than those of either parent, and there was a marked difference between reciprocal crosses. Synaptonemal complexes of homozygous females appeared as 19 bivalents, those of single heterozygotes as 18 bivalents and a trivalent, and those of compound heterozygotes as 17 bivalents and a quadrivalent. Most trivalents were fully paired, whereas the majority of quadrivalents exhibited terminal asynapsis. About one-half of all oocytes had other pairing abnormalities, probably reflecting reduced survivability. Whereas all fully paired quadrivalents were present in cells not showing any pairing anomalies, one-half of the quadrivalents with terminal asynapsis were seen in oocytes with other anomalies. It is suggested that in oocytes destined for atresia, there is a predisposition to synaptic failure of translocation configurations. Additional oocytes are likely to break down because of the deleterious effect of the compound translocation on gametogenesis. This effect seems to be more pronounced in Rb1Ald/Rb2Bnr spermatocytes than in oocytes.     

Pachytene analysis of a man with a 13q;14q translocation and infertility. Behavior of the trivalent and nonrandom association with the sex vesicle

Pachytene analysis was undertaken in a sterile 13q;14q heterozygous translocation carrier in an attempt to follow the segregational behavior of the trivalent and to evaluate the relationship of Robertsonian translocations in man to the impairment of spermatogenesis. Well-spread bivalents from pachytene nuclei were identified by their chromomere patterns. The trivalent was found always in cis configuration. Silver staining demonstrated the loss of nucleolar organizer regions from the translocated chromosomes. A nonrandom association was found between the trivalent configuration and the sex vesicle in 61% of the pachytene nuclei examined. Such an association has been described before in mice heterozygous for Robertsonian or reciprocal translocations, and may thus represent a general phenomenon. As in mice, this contact was restricted to the centromeric region of the trivalent. A hypothesis relating the association of the trivalent with the sex vesicle to impairment of normal X-chromosome inactivation and subsequent spermatogenic breakdown is discussed. Other chromosomal abnormalities in which sex-vesicle anomalies are associated with male sterility (such as X-or Y-autosomal translocations) are also considered. It is proposed that any process interfering with normal X-chromosome inactivation in pachytene spermatocytes could disturb subsequent meiotic or postmeiotic germ cells development.     

Male meiosis and gametogenesis in wild house mice (Mus musculus domesticus) from a chromosomal hybrid zone; a comparison between "simple" Robertsonian heterozygotes and homozygotes.

Wild male house mice Mus musculus domesticus were collected from the hybrid zone between the John o'Groats race (2n = 32) and the standard race (2n = 40) in northern Scotland. Meiosis in both homozygotes (2n = 32, 36, and 40) and single Robertsonian heterozygotes (2n = 33, 35, and 37) was found to be orderly. At prophase/metaphase I in heterozygotes, a trivalent was formed from the metacentric and two homologous acrocentrics. At pachytene, this trivalent usually had a single side arm at the position of the centromeres, as a result of nonhomologous pairing of the acrocentrics. This side arm persisted into diplotene. Generally only a single chiasma was formed between each acrocentric and the metacentric. Anaphase I nondisjunction frequencies were estimated as 1.5% for the homozygotes and 2.7% for the heterozygotes. The extent of germ cell death between the pachytene and round spermatid stages was 18% greater in heterozygotes than in homozygotes. Our results concur with previous studies which indicate that single Robertsonian heterozygotes in wild house mice have near-normal fertility.     

Further examination of the production-line hypothesis in mouse foetal oocytes

Pachytene oocytes from foetal mice heterozygous for the translocation T(14; 15)6Ca were screened for evidence of a production-line effect on chromosome pairing. Metaphase I oocytes from adult heterozygotes were also examined to determine whether any such effect on pahytene chromosome pairing is subsequently repeated during adult reproductive life as anticipated by the production-line hypothesis. It was found that as gestation proceeded the proportion of pachytene oocytes with a translocation quadrivalent declined and that with a trivalent and univalent correspondingly increased. That is, there was evidence of variation in pairing behaviour of the translocation at different times of gestation. In contrast, the proportions of metaphase I cells with either a quadrivalent or a trivalent plus univalent did not vary between adult females of different ages. Thus if the variation observed at pachytene was the result of a production-line effect, clearly this was not reflected in the behaviour of the translocation at metaphase I. Our observations therefore do not support the production line hypothesis for the maternal age effect on nondisjunction.     

Two new X-autosome Robertsonian translocations in the mouse

The influence of X-autosome Robertsonian (Rb) translocation hemizygosity on meiotic chromosome behaviour was investigated in male mice. Two male fertile translocations [Rb(X.2)2Ad and Rb(X.9)6H] and a male sterile translocation [Rb(X.12)7H] were used. In males of all three Rb translocation types, the acrocentric homologue of the autosome involved in the rearrangement regularly failed at pachytene to pair completely with its partner in the Rb metacentric. The centric end of the acrocentric autosome was found regularly to associate either with the proximal end of the Y chromosome or with the ends of nonhomologous autosomal bivalents; the proportions of cells with such configurations varied between pachytene substages and genotypes. Various other categories of synaptic anomaly, such as nonhomologous synapsis, foldback pairing and interlocks, affected the sex chromosome multivalent in a substantial proportion of cells. In one of the Rb(X.12)7H males screened, an unusual, highly aneuploid spermatocyte that contained trivalent and bivalent configurations was found. Rb translocation hemizygosity did not appear to increase to a significant extent the incidence of X-Y pairing failure at pachytene, although the incidence was elevated at metaphase I in Rb(X.12)7H animals. Overall, a comparison of the frequencies and types of chromosome pairing anomalies did not suggest that these were important factors in the aetiology of infertility in males carrying the Rb(X.12)7H translocation.     

Pachytene pairing and sperm counts in mice with single Robertsonian translocations and monobrachial compounds

Data on testis weights, sperm counts, and synaptonemal complexes are presented for mice carrying the following Robertsonian translocations: Rb(6.15)lAld; Rb(4.6)2Bnr; Rb(4.15)4Rma; Rb(6.15)lAld/Rb(4.6))2Bnr, which is male-sterile; and Rb(6.15)lAld/Rb(4.15)4Rma, which is male-fertile. In RblAld/Rb2Bnr sperm were absent or sparse, whereas the sperm count in RblAld/Rb4Rma was just over 50% of the parental value. The translocated chromosomes appeared as fully paired bivalents in homozygotes, as trivalents in single heterozygotes, and as quadrivalents in compounds. About 20-40% of trivalents had unsynapsed ends. The proportion of quadrivalents with unsynapsed ends was about 85% in the male-sterile compound, compared with 75% in the male-fertile compound. The proportion of quadrivalents associated with XY was about 70% in both. Testis weights, but not sperm counts, were found to differ in two of three reciprocal crosses. It is concluded that, in addition to pairing failure and autosome/XY association, the effect of translocations on spermatogenesis is affected by other factors, including genetic background, inbreeding, and perhaps environmental factors. It remains to be elucidated whether pairing failure and XY association are primary or secondary effects.     

Orientation and segregation of Robertsonian trivalents in Dichroplus pratensis (Acrididae)

Pairing behavior, metaphase I orientation, and anaphase I segregation of centric fusion trivalents were studied in 26 single, 15 double, and 2 triple male fusion heterozygotes of the polymorphic South American melanopline grasshopper Dichroplus pratensis. They represent the seven different fusions and their combinations already described in different populations of the species. Our analysis showed the following: (1) pairing behavior is very regular in all trivalents; (2) frequencies of linear orientation was very low irrespective of the trivalent involved; (3) reorientation seems to occur frequently since frequencies of abnormal segregation and aneuploid second division cells were invariably lower than those of nonconvergent orientation; (4) aneuploidy and abnormal sperm production increases with increasing number of fusions; (5) chiasma frequency and localisation is relevant to trivalent orientation since trivalents with nonconvergent orientations showed proximal and interstitial chiasmata more frequently than convergently oriented ones. The results are in agreement with the hypothesis that these polymorphisms are old and stable, and confirm that for the maintenance of a balanced polymorphism, if this polymorphism is adaptive because of its consequences on recombination, position effects, etc., changes tending to stabilise trivalent orientation and segregation are central.     

Further examination of the production-line hypothesis in mouse foetal oocytes

Chromosome pairing has been examined in foetal oocytes of mice heterozygous either for an X-linked inversion, In(X)1H, or an autosomal inversion, In(2)2H. The patterns of chromosome pairing have been screened systematically in foetuses of different gestational ages in a search for a production-line effect particularly affecting the inversion-bearing bivalents. The proportion of pachytene oocytes with a loop fell with increasing gestational age for both inversions. The decrease was linear for In(X)1H but best described by a quadratic function for In(2)2H. Examination of late zygotene cells and a comparison of loop frequency in early, mid and late pachytene oocytes suggested this age-related decrease to be principally due to synaptic adjustment and not to a production-line effect. However, two particular observations were somewhat at variance with this conclusion. Firstly, in In(X)1H heterozygotes, the presence of an inversion loop and the occurrence of partial pairing of long/long-medium bivalents at pachytene were independent of each other only on day 19. Secondly, although the proportion of oocytes with a loop fell overall, there was a rise at 19 days in In(2)2H heterozygotes. Thus in both inversions there is some evidence of a change in pairing behaviour affecting the inversion-bearing bivalents at the latest gestational age, as would be expected under the production-line hypothesis.     

Chromosomal evolution in Malagasy lemurs. II. Meiosis in intra- and interspecific hybrids in the genus Lemur.

The chromosome analysis of meiosis in four lemurs, Lemur fulvus fulvus, L. f. collaris, L. f. albocollaris, and L. macaco, and particular hybrid crosses is reported. In metaphase I, trivalents and chain elements were detected and identified with T-banding. The study largely confirms our previous work on mitotic karyotype comparisons in the genus Lemur. The absence of chain multivalents elements in the pachytene stages of hybrid meiosis, where a chain is detected later in diakinesis, may offer evidence on the possible existence of a two-step pairing mechanism in meiotic homolog pairing. Considerations about the role of the chromosome rearrangements in establishing a gametic barrier in speciation are developed.