Chromosome segregation in mice heterozygous for Robertsonian translocations. I. The effect of mutation in chromosome 17 on the disjunction of homologs in females

The effects of mutations on chromosome 17 upon the segregation of the metacentric and acrocentric homologues in the progeny of female mice heterozygous for Robertsonian translocations Rb(8.17) 1Iem and Rb(16.17) 7Bnr were studied. Genetic analysis indicated that the ratio of Rb to non-Rb (normal karyotype) progeny from mothers heterozygous for mutations tf, qk, t12 were not significantly different from 1:1 expected. Introduction of mutations T, Ki, Fu, t6 into the female genotype caused strong distortion of segregation and an increase in the proportion of progeny with normal karyotype (65-70%). From the data on embryonic mortality and cytogenetic observations, it is concluded that distortion of equal transmission arises before MII of meiosis. Consequently, preferential distribution of the metacentric chromosome into the polar body during the first meiotic division is relevant to the segregation distortion observed.     

Gamete segregation in female carriers of Robertsonian translocations

Eleven female carriers of either 45,XX,der(13;14) (q10;q10) or 45,XX, der(14;21)(q10;q10) underwent hormonal stimulation with the purpose of producing enough oocytes for in-vitro fertilization and preimplantation genetic diagnosis. Polar body biopsy was performed in those oocytes and FISH with painting probes was applied in their metaphase-like first polar body chromosomes. In this way, unbalanced, normal and balanced oocytes could be distinguished and segregation modes ascertained. der(14;21)(q10;q10) produced 42% unbalanced, 37% normal and 21% balanced oocytes (n = 86) while der(13;14)(q10;q10) generated 33% unbalanced, 51% normal and 16% balanced oocytes (n = 69). In both translocations the number of normal oocytes was significantly higher than the number of balanced oocytes. However, while the frequency of unbalanced events involving chromosome 13 and 14 was similar in der(13;14)(q10;q10), there were significantly more abnormalities involving chromosome 21 than 14 in the der(14;21) (q10;q10) cases. When comparing survival rates to term, trisomies from Robertsonian origin seem to survive more often than those originated by non-disjunction in non-translocation carriers. The meiotic segregation patterns found in female Robertsonian translocations are different from those described in male carriers, with higher rates of unbalanced gametes in females than in males.     

Kinetics of oogenesis in mice heterozygous for Robertsonian translocations

The total number of oocytes at different postmating time intervals (18-40 days) was determined in mice homozygous and heterozygous for different Robertsonian (Rb) translocations, of both laboratory and feral origin. The number of oocytes was lower in heterozygous than in homozygous mice throughout the period studied. Independently of the genetic background (i.e. laboratory or feral), structural heterozygosity had a progressive detrimental effect on oocyte numbers: open, or chain diakinetic configurations had a greater detrimental effect than close, or ring, configurations. The genetic background, however, affected the ovarian constitution in terms of the total number of germ cells, which are more numerous in laboratory than in feral mice. The kinetics of oogenesis seems to be faster in feral than in laboratory mice. At the light of the data here presented, and of those already available from the literature on male and female gametogenesis in conditions of structural heterozygosity, it appears that factors other than unsaturation of pairing sites or interference with pachytene X-chromosome inactivation have to be considered. In the wild, the reduced oocyte numbers in Rb heterozygous female can contribute to the retention of isolated populations in contact zones.     

Transmission ratio distortion in offspring of heterozygous female carriers of Robertsonian translocations

Robertsonian translocations are the most common structural rearrangements of human chromosomes. Although segregation of Robertsonian chromosomes has been examined in many families, there is little consensus on whether inheritance in the balanced progeny conforms to Mendelian ratios. To address this question, we have compiled previously reported segregation data, by sex of parent, for 677 balanced offspring of Robertsonian carriers from 82 informative families and from a prenatal diagnosis study on the risk of unbalanced offspring in carriers of chromosome rearrangements. Care was taken to avoid any source of ascertainment bias. Our analysis supports the following conclusions: (1) the transmission ratio is not independent of the sex of the carrier; (2) the transmission ratio distortion is observed consistently only among the offspring of carrier females; (3) the transmission ratio distortion does not appear to be dependent on the presence of a specific acrocentric chromosome in the rearrangement. The sex-of-parent-specific origin of the non-Mendelian inheritance, the finding that the rearranged ("mutant") chromosomes are recovered at significantly higher frequency than the acrocentric ("normal") chromosomes, and the similarities between these observations and the segregation of analogous rearrangements through female meiosis in other vertebrates strongly support the hypothesis that the transmission ratio distortion in favor of Robertsonian translocations in the human results from the preferential segregation of chromosomes during the first meiotic division. This non-Mendelian inheritance will result in increased overall risk of aneuploidies in the families of Robertsonian translocation carriers, independently of the origin of the transmission ratio distortion.     

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.     

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.     

Nondisjunction, disturbances in spindle structure, and characteristics of chromosome alignment in maturing oocytes of mice heterozygous for Robertsonian translocations

To correlate the chromosomal constitution of meiotic cells with possible disturbances in spindle function and the etiology of nondisjunction, we examined the spindle apparatus and chromosome behavior in maturing oocytes and analyzed the chromosomal constitution of metaphase II-arrested oocytes of CD/Cremona mice, which are heterozygous for a large number of Robertsonian translocation chromosomes (18 heterobrachial metacentrics in addition to two acrocentric chromosomes 19 and two X chromosomes). Spreading of oocytes during prometaphase 1 revealed that nearly all oocytes of the heterozygotes contained one large ring multivalent, apart from the bivalents of the two acrocentric chromosomes 19 and the X chromosomes, indicating that proper pairing and crossing-over between the homologous chromosome arms of all heterobrachial chromosomes took place during prophase. A large proportion of in vitro-matured oocytes arrested in metaphase II exhibited numerical chromosome aberrations (26.5% hyperploids, 40.8% hypoploids, and 6.1% diploids). In addition, some of the oocytes with euploid chromosome numbers (26.5% of the total examined) appeared to be nullisomic for one chromosome and disomic for another chromosome, so that aneuploidy levels may even be higher than expected on the basis of chromosome counts alone. Although oocytes of the complex heterozygous mice seemed able initially to form a bipolar spindle during first prometaphase, metaphase I spindles were frequently asymmetrical. Chromosomes in the multivalent did not align properly at the equator, centromeres of neighboring chromosomes in the multivalent remained maloriented, and pronounced lagging of chromosomes was observed at telophase I in oocytes obtained from the Robertsonian translocation heterozygotes. Therefore, disturbance in spindle structure and chromosome behavior appear to correlate with the chromosomal constitution in these oocytes and, ultimately, with failures in proper chromosome separation. In particular, reorientation appears to be a rare event, and malorientation of chromosomes may remain uncorrected throughout prometaphase, as we could not find many typical metaphase I stages in heterozygotes. This, in turn, could be the basis for malsegregation at anaphase and may ultimately induce a high rate of nondisjunction and aneuploidy in the oocytes of CD/Cremona mice, leading to total sterility in heterozygous females.     

Two new X-autosome Robertsonian translocations in the mouse. I. Meiotic chromosome segregation in male hemizygotes and female heterozygotes.

Two new X-autosome Robertsonian (Rb) translocations, Rb(X.9)6H and Rb(X.12)7H, were found during the course of breeding the Rb(X.2)2Ad rearrangement at Harwell. The influence of these new Rbs on meiotic chromosome segregation was investigated in hemizygous males and heterozygous females and compared to that of Rb(X.2)2Ad. Screening of metaphase II spermatocytes gave incidences of sex chromosome aneuploidy of 9.2% in Rb(X.2)6H/Y and 9.6% in Rb(X.9)2Ad/Y males; no metaphase II cells were present in the testes of the Rb(X.12)7H/Y males examined and no males with this karyotype have so far proved fertile. In breeding tests, 5% of the progeny of Rb(X.2)2Ad/Y males were sex chromosome aneuploids compared to 10% of the Rb(X.9)6H/Y offspring. The difference was not significant, however. Cytogenetic analyses of metaphase II stage oocytes showed elevated rates of hyperhaploidy (n + 1) in Rb heterozygous females over chromosomally normal mice: 4.2% for Rb(X.2)2Ad/+; 2.1% for Rb(X.9)6H/+; 2.2% for Rb(X.12)7H/+ and 1.1% for normal females. There was, however, no statistically significant difference in the rates of hyperhaploidy between the three different Rb types, nor overall between Rb/+ and normal females. Karyotypic analyses of liveborn offspring of Rb heterozygous females revealed low incidences of X0 animals but no other type of sex chromosome aneuploidy. Intercrosses of heterozygous females and hemizygous males yielded 5.5% aneuploidy for Rb(X.2)2Ad and 5.4% for Rb(X.9)6H. In heterozygous females, there was evidence from the metaphase II and breeding test data for all three rearrangements, of preferential segregation of the Rb metacentric to the polar body resulting in a deficiency of cells and progeny carrying a translocation chromosome.     

Chromosome segregation in mice heterozygous for Robertsonian translocations. II. Changes in the structure of homologs as a cause of abnormal disjunction in females

It was demonstrated that mutations T, Fu, Ki, t6 of chromosome 17 cause preferential transmission of the acrocentric homologues to the progeny from female Rb heterozygotes. The results indicate that the effects of these mutations on segregation are restricted to the Robertsonian translocations involving chromosome 17. Substitution of the parts of chromosome 17 distal or proximal to the T-locus did not alter the effect, of this chromosome on the transmission rate of the homologue. The transmissions effects of these mutations, whether cis or trans with Rb, were the same. It was observed that mothers Rb7/T43H transmitted the chromosome with the reciprocal translocation T43H to 70.9% of their progeny. Data were obtained supporting the idea that structural changes of the chromosomes caused by mutations affect segregation of the homologues in Rb heterozygous females. The possible mechanism of this influence is discussed.     

Chiasma distributions and chromosome segregation in male and female translocation heterozygous mice analysed using FISH

Two factors postulated to influence the meiotic behaviour of reciprocal translocations were investigated. Firstly, variation in the length of translocated and non-translocated segments was studied in male mice using four different rearrangements involving chromosomes 2 and 4. Secondly, sex-related effects were analysed through comparison of the meiotic behaviour of two translocations in male and female germ cells. In the first part of the study, primary and secondary spermatocytes of male mice carrying a translocation [T(2;4)1Ca, T(2;4)13H, T(2;4)1Sn, or T(2;4)1Go] were screened. Each rearrangement had different proportions of cells with ring and chain quadrivalents at metaphase I; the T(2;4)1Sn heterozygote also had a high rate (45%) of translocation bivalents. In general, the translocations had elevated chiasma frequencies in the rearranged chromosomes compared with structurally normal chromosomes 2 and 4, although the extent of the effect varied. Each rearrangement produced a different array of segregation products at metaphase II, reflecting their contrasting frequencies of multivalent configurations at metaphase I. Comparison of chromosome behaviour at metaphase I and II suggested that certain configurations tended to adopt particular orientations. However, it was also clear that such correlations were imprecise and that other factors, possibly the exact positions of chiasmata, also played a role in multivalent orientation. Two rearrangements, T(2;4)1Go and T(7;16)67H, were analysed in female mice. The frequencies of the various multivalent types at metaphase I differed from those in male carriers of these rearrangements owing to an increased chiasma frequency in oocytes in some of the pairing segments. Not surprisingly, the segregation products seen in metaphase II cells showed some differences from the pattern recorded in male germ cells. For T(2;4)1Go, the sex-related difference in segregation patterns resulted in a diminished expectation of genetically imbalanced gametes, although this was not the case for T(7;16)67H. Received: 6 June 1998 / Accepted: 9 October 1998     

Meiotic exchange and segregation in female mice heterozygous for paracentric inversions

Inversion heterozygosity has long been noted for its ability to suppress the transmission of recombinant chromosomes, as well as for altering the frequency and location of recombination events. In our search for meiotic situations with enrichment for nonexchange and/or single distal-exchange chromosome pairs, exchange configurations that are at higher risk for nondisjunction in humans and other organisms, we examined both exchange and segregation patterns in 2728 oocytes from mice heterozygous for paracentric inversions, as well as controls. We found dramatic alterations in exchange position in the heterozygotes, including an increased frequency of distal exchanges for two of the inversions studied. However, nondisjunction was not significantly increased in oocytes heterozygous for any inversion. When data from all inversion heterozygotes were pooled, meiotic nondisjunction was slightly but significantly higher in inversion heterozygotes (1.2%) than in controls (0%), although the frequency was still too low to justify the use of inversion heterozygotes as a model of human nondisjunction.     

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.     

Meiotic drive in female mice heterozygous for the HSR inserts on chromosome 1

Chromosome 1 with one or two long insertions has been previously found in natural mouse populations. The inheritance of chromosome 1 with two insertions from the Yakutsk population is analysed in this paper. It was demonstrated that heterozygous females transmit this chromosome to 80-85% of offspring. The observations made at M II, in conjunction with the recombination data, allowed us to conclude that preferential passage of the chromosome 1 with insertions to the oocyte and egg, rather than to the first and second polar bodies at meiosis, is the causative factor of the distorted segregation. A meiotic drive of such potency has not been previously reported for female mammals. The possible mechanism of the drive is discussed.     

Meiosis in trisomic female mice with Robertsonian translocations. I. Prophase pairing

The prophase oocytes of two murine Robertsonian translocation (Rb) trisomies of chromosomes 16 and 19 were investigated using electron microscopy and a whole-cell micro-spreading technique after silver staining. About 20% of fetuses of each type were trisomic. They were obtained by mating animals heterozygous for two Rb's, monobrachially homologous for either chromosome 16 or 19, to an entirely acrocentric stock. Because of the almost inevitable prenatal mortality of the trisomic embryos, their fetal ovaries were "rescued" by an in vitro method for prophase studies. Analysis of the recovered oocytes showed frequent, close pairing associations of the three trisomic axes and evidence suggesting that the closely apposed axes coincided with the side-by-side formation of parallel, complete, true synaptonemal complexes; hence, the cytogenetic dogma that pairing is always two-by-two was contradicted. The presence of two parallel complexes has implications for crossing-over recombination. Triple associations of axes were found in almost half the trisomy 19 (Ts19) and in about 70% of the trisomy 16 (Ts16) prophases. The extent of triple associations varied and was greater in Ts16 than in Ts19 oocytes. Other relevant observations concerned the proportions of univalents and of univalence of the trisomic axes (21% in Ts16 and 46% in Ts19) and the distinctive, thickened appearance of all univalent axes. The pairing behaviour observed in balanced heterozygotes confirms what appears to be nonhomologous pairing and synaptic adjustment within the short-arm axes of the Rb trivalents.     

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.     

Chromosome pairing and recombination in mice heterozygous for different translocations in chromosomes 16 and 17

In order to clarify the relationship between meiotic pairing and recombination, and electron microscopic (EM) study of synaptonemal complexes (SC) and an analysis of chiasma frequency and distribution were made in male mice singly and doubly heterozygous for Robertsonian [Rb(16.17)7Bnr] and reciprocal [T(16:17)43H] translocations and also in tertiary trisomics for the proximal region of chromosome 17. In all these genotypes an extensive zone of asynapsis/desynapsis around the breakpoints was revealed. At the same time a high frequency of non-homologous pairing was observed in precentromeric regions of acrocentric chromosomes. The presence in the proximal region of chromosome 17 of the t haplotype did not affect the synaptic behaviour of this region. Chiasma frequency in the proximal region of chromosome 17 in the T(16:17)43H heterozygotes and trisomics was increased when compared with that in Robertsonian heterozygotes.by H.C. Macgregor     

Developmental stability in house mice heterozygous for single Robertsonian fusions

Fluctuating asymmetry (FA) of tooth traits has been reported to be increased in Down syndrome patients as well as hybrids between chromosomal races of the house mouse differing in several Robertsonian (Rb) fusions. Developmental stability, assessed by FA, is thus thought to be impaired by spontaneous chromosomal abnormality or by chromosomal heterozygosity. Although the effect of a single fusion on developmental stability could theoretically be expected, it has never been documented. Crosses involving two chromosomal races of the house mouse diverging for one Rb fusion were performed to assess developmental stability in parental homozygous races as well as in their hybrids. Moreover, the occurrence of a spontaneous chromosomal mutation (WART type-b) allowed us to study the instantaneous effect of such a translocation on developmental stability. No difference in fluctuating asymmetry levels was detected among the groups considered in this study. This result suggested that a single stable or spontaneous balanced structural rearrangement did not inherently disturb developmental stability. In addition, the differential effect on developmental stability of one versus many heterozygous Rb fusions highlights the role of their quantitative accumulation in the disruption of coadaptation in chromosomal hybrids.