From spermatocytes to sperm: meiotic behaviour of human male reciprocal translocations

M. Oliver-Bonet1,3, J. Navarro1, M. Codina-Pascual1, C. Abad2, M. Guitart2, J. Egozcue1 and J. Benet1,3

1 Unitat de Biologia, Facultat de Medicina, Departament de Biologia Cel.lular, Fisiologia i d'Immunologia, Universitat Autònoma de Barcelona, Bellaterra and 2 Consorci Hospitalari Parc Taulí, Sabadell, Spain

3 To whom correspondence should be addressed at: Unitat de Biologia, Facultat de Medicina, Edifici M, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain. E-mail: or Email: moliver{at}servet.uab.es


    Abstract
 Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Acknowledgements
 References
 
BACKGROUND: Human male translocation carriers may present alterations in the meiotic process due to the presence of the translocated chromosomes. The aim of this work was to study the mechanisms that affect meiotic segregation in translocation carriers by analysing different stages of the meiotic process. METHODS: Meiotic studies using fluorescence in-situ hybridization on both spermatocytes and sperm nuclei were performed in two translocation carriers, t(11;17)(q13.1;p11.2) and t(10;14)(q24;q32). RESULTS: A ring configuration was the main type of quadrivalent found in metaphase I. Overall chiasma frequency was significantly decreased in the t(11;17) carrier. In the t(10;14) carrier, chiasma frequency within the interstitial region of chromosomes 10 and 14 was increased and the recombination pattern was modified. As expected from the frequencies of interstitial chiasmata found in metaphase I in the two subjects, the incidence of asymmetric dyads was sporadic in t(11;17) and very high in t(10;14). In both carriers, segregation frequencies observed at metaphase II were not different from the segregation data obtained in decondensed sperm nuclei. CONCLUSIONS: The concordance observed among results obtained in different spermatogenic stages indicates an absence of cellular selection based on chromosomal imbalances. Results obtained in the aneuploidy assay have not provided any evidence for an interchromosomal effect.

Key words: chromosome rearrangements/FISH/meiotic segregation/sperm/spermatocytes


    Introduction
 Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Acknowledgements
 References
 
Meiosis is the process by which haploid gametes are produced. It includes two successive cell divisions, after just one round of DNA replication. The pairing of homologous chromosomes (meiotic synapsis) and the exchange of genetic information between them (chromosome recombination) takes place during early prophase of the first meiotic division. Both synapsis and meiotic recombination play a critical role in meiosis. When the proteinaceuos structure that holds synaptic processes (synaptonemal complex) disappears during diplotene, homologous chromosomes are held together by crossover events. If recombination fails, homologous segregation is disturbed, and the bivalents become univalents (Egozcue et al., 2000Go). The behaviour of the univalents is not predictable, and the risk of generating chromosomal abnormalities due to non-disjunction, anaphase lag or misdivision during meiosis I is increased (Hassold and Hunt, 2001Go). Some of these abnormalities will not affect the viability of the meiotic cell, and the meiotic process will be completed, in which case a chromosomally abnormal gamete will be formed. This can have clinical consequences. In fact, nearly half of spontaneous abortions in clinically reorganized pregnancies are chromosomally abnormal (Hassold et al., 1996Go).

Structural chromosomal rearrangements are present in 2% of spontaneous abortions (Jacobs, 1990Go), and most structural abnormalities (80%) are inherited from one of the parents (Jacobs, 1992Go), who carries a balanced chromosomal rearrangement. The presence of a reorganization may alter the normal progress of meiosis in two different ways. The first is related to the reorganization itself. In the case of a reciprocal translocation, the chromosomes involved produce a quadrivalent during the synaptic process. The behaviour of this particular structure during meiotic segregation is unique for each translocation carrier (Cifuentes et al., 1999Go): it determines the proportion of balanced and unbalanced gametes for the carrier tested, depends upon several factors, such as the size of the interstitial and translocated segments, the morphological characteristics of the rearranged chromosomes and the distribution pattern of chiasmata during metaphase I (MI), and affects the final frequencies of normal and aberrant gametes. The proportion of balanced and unbalanced phenotypes differs greatly from one carrier to another (Guttenbach et al., 1997Go; Shi and Martin, 2001Go). Information about the presence of chiasmata and, in particular, interstitial chiasmata in the quadrivalent can help to elucidate the segregational mechanisms which lead to the production of different proportions of balanced and unbalanced gametes (Armstrong and Hultén, 1998Go).

The second way in which a reorganization may alter meiosis is a phenomenon known as the interchromosomal effect (ICE) (Lejeune, 1963Go). In this case, the presence of a structural abnormality may disturb the meiotic segregation of other chromosome pairs not related to the rearrangement, thus producing aneuploid sperm. Then, ICE can be evaluated through an increase of the aneuploidy frequencies of other chromosomes, which are not involved in the reorganization (Estop et al., 2000Go).

To understand the mechanisms that affect meiotic segregation patterns in human male carriers of reciprocal translocation, the direct analysis of MI and metaphase II (MII) in testicular biopsies is necessary. In this study, fluorescence in-situ hybridization (FISH) was used on meiotic spreads of two translocation carriers, t(11;17)(q13.1;p11.2) and t(10;14)(q24;q32). This method allowed the identification of the MI quadrivalent, the determination of chiasma frequencies in the quadrivalent and in other bivalents, and the detection of balanced and unbalanced segregating products at MII. Finally, meiotic products may be targeted by FISH on decondensed sperm nuclei. The proportion of each different sperm phenotype has been determined in order to compare it with the segregation pattern obtained by the analysis of MII. Finally, the presence of possible ICE for chromosomes X, Y, 6, 18 and 21 was also tested. This is, to our knowledge, the first time that reciprocal translocations have been analysed at different stages of meiosis.


    Materials and methods
 Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Acknowledgements
 References
 
Donors
One patient had a 46,XY,t(11;17)(q13.1;p11.2) karyotype. He was ascertained because of repeated reproductive failures. The other patient carried a t(10;14)(q24;q32). He was also ascertained because of repeated reproductive failures that ended in miscarriage. According to WHO parameters, both carriers had normal semenograms (World Health Organization, 1999Go). Ideograms for both the normal and the translocated chromosomes, as well as quadrivalent configurations at meiosis I, are shown in Figure 1.



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Figure 1. Ideograms of t(11;17)(q13.1;p11.2) and t(10;14)(q24;q32).

 
Two normozoospermic males (aged 25 and 31 years at the time of sample collection) were used as a control for the sperm FISH analysis.

Testicular biopsies were obtained from both patients by open incision under local anaesthesia. Each patient provided one semen sample prior to surgery. Fresh ejaculates were allowed to liquefy and then aliquoted and cryopreserved. This study was approved by the institutional ethics committee, and the patients gave written informed consent.

Meiotic cell extensions and FISH treatment
Testicular tissue was placed in a hypotonic solution (sodium citrate 1%) until the tubules were disaggregated to obtain a cell suspension, and then placed in a centrifuge tube to separate testicular cells from seminiferous tubule remnants. The supernatant was recovered and placed in another tube, centrifuged for 5 min at room temperature at 600 g, and finally resuspended in a few drops of sodium citrate 1%.

Fixation and spreading of the cells was performed using the method described elsewhere (Evans et al., 1964Go). Each slide was screened by phase-contrast microscopy, and phase images of each chromosome extension at 40x magnifications were captured and stored using a Cytovision imaging system (Applied Imaging, Sunderland, UK). Slides were then counterstained with DAPI (4',6-diamino-2-phenyndole), and images of the previously captured extensions were obtained at 100x and stored using the same system.

Meiotic chromosome extensions were hybridized with probe mixtures containing whole chromosome painting for the translocated chromosomes and centromeric probes for the sex chromosomes (Table I). Hybridizations were carried out according to the procedure described previously (Oliver-Bonet et al., 2001Go) with some modifications: prior to postfixation, DAPI was removed from the slides using a 4x standard sodium citrate (SSC)/0.05% Tween 20 solution (5 min), followed by an ethanol series (70%, 85%, 100%). Five microlitres of probe mix were added to each slide and co-denaturation was performed on a hotplate heated to 68°C for 2 min. Co-incubation was performed overnight at 37°C in a humidified chamber. Then, slides were washed in 0.4x SSC at 70°C for 2 min, followed by 2 min in 4x SSC/0.05% Tween 20. Finally, slides were counterstained with antifade (Vector Laboratories Inc., Burlingame, CA, USA) containing DAPI at a concentration of 0.032 ng/ml (Sigma, Madrid, Spain).


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Table I. DNA probes used for the three different analyses

 
Sperm nuclei extension and FISH treatment
To obtain sperm nuclei, an aliquot of frozen spermatozoa was thawed and washed in NaCl 0.9% to eliminate cryoprotectant. The sample was fixed and decondensed following a previously described procedure (Vidal et al., 1993Go).

Decondensed sperm nuclei extensions were hybridized for the segregation analysis using a combination of specific probes for the centromeric and telomeric regions of the chromosomes implicated in the reorganization. Aneuploidy studies using two- and three-colour FISH were performed also on decondensed sperm nuclei in order to detect any possible ICE. Probes used are described in Table I. Denaturation and hybridization were performed following a procedure described elsewhere (Oliver-Bonet et al., 2001Go). Slides were incubated overnight at 37°C, and then washed following the manufacturer's instructions, dehydrated and counterstained with antifade (Vector Laboratories Inc.) containing DAPI at a concentration of 0.032 ng/ml (Sigma).

Image capture
Visualization of FISH signals was made using an Olympus Ax 70 photomicroscope (Olympus Optical Co., Hamburg, Germany) equipped with four simple filters for visualizing DAPI, FITC, Aqua and Cy3 fluorescence, and a fifth triple filter for DAPI/FITC/PI. The capture and analysis of FISH images were performed using the Cytovision FISH imaging system (Applied Imaging) Sunderland, UK.

Cytogenetic analysis
Both contrast and DAPI images were used to asses chiasmata in MI (Figure 2A and B). Data obtained from male subjects with normal karyotype were used as control (Laurie and Hultén, 1985Go). FISH analysis allowed the identification of the chromosomes implicated in the reorganization. Localization of chiasmata within the quadrivalent was defined in paired terminal region as pt or qt, in not terminal regions as p(short arm) or q(long arm), and in interstitial segments as qint or pint (Figure 1). In MII cells, normal, rearranged and dimorphic chromosomes were defined after FISH analysis.



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Figure 2. (a) Contrast and FISH images of a MI spread of t(11;17)(q13.1;p11.2). A ring configuration with five chiasmata, including an interstitial chiasma in 17p (red arrow), can be seen. Whole chromosome painting for chromosome 11 (green) and 17 (red); centromere signal for X (green) and Y (red) chromosomes. (b) Contrast and FISH images of a MI spread of t(10;14)(q24;q32). In this case, the ring has one interstitial chiasma in 10q and another in 14q (red arrows). Whole chromosome painting for chromosome 10 (green) and 14 (red); centromere signal for X (green) and Y (red) chromosomes.

 
Evaluation of FISH signals in decondensed sperm nuclei was made following standard criteria as described previously (Egozcue et al., 1997Go).

Statistical methods
Student's t-test and the {chi}2-test were used for data analysis.


    Results
 Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Acknowledgements
 References
 
Chiasma analysis on MI
Results for the t(11;17) carrier are summarized in Table II. All configurations involving the translocated chromosomes in MI were quadrivalents, and most of them (91.8%) were rings. In 8.2%, a chain configuration was found, with only three chiasmata and a lack of chiasma formation at 11q (3.3%) or 17p (4.9%). The number of chiasmata found within quadrivalents with a ring configuration ranged from four to six, with at least one chiasma per chromosome arm of each translocated chromosome. Nearly half (44.3%) of the ring quadrivalents contained four terminal chiasmata. Configurations with five chiasmata were found in 42.6% of the quadrivalents. Only 1.6% of them had the fifth chiasmata located in 17pint (Figure 2A), while the rest had this fifth chiasmata located in 11q (27.9%), 17q (9.8%) or 11p (3.3%). Quadrivalents with six chiasmata represented 4.9% of the total number, and the fifth and sixth chiasmata were situated one in 11q and the other in 17q.


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Table II. Distribution of different configurations for the t(11;17) at MI

 
In the t(10;14) carrier, ring configurations with four, five or six chiasmata were found in 96.1% of the quadrivalents. A chain configuration, with either three or four chiasmata, was observed in 3.9% of the spreads. Frequencies of the different configurations are summarized in Table III. One single interstitial chiasma in chromosome 10 was present in 78.4% of the quadrivalents. Interstitial chiasma in chromosome 14 ranged from between zero and two, and frequencies of quadrivalents with one and two interstitial chiasmata in this chromosome were 70.6% and 23.5%, respectively (Figure 2B).


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Table III. Distribution of different configurations for the t(10;14) at MI

 
The mean chiasma frequencies were calculated for each arm of the translocated chromosomes, as well as for the interstitial segments, in both translocation carriers (Table IV). Then, these results were compared with data obtained from male subjects with a normal karyotype (Laurie and Hultén, 1985Go). We observed a significant increase in chiasma frequencies in 10q (P=0.003) and 14q (P=0.004).


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Table IV. Chiasmata frequency per chromosome arm and for all autosomes

 
Chiasmata were also counted in all bivalents at MI stage. The mean [±standard deviation (SD)] chiasma frequency recorded per cell in the t(11;17) carrier was 48.9±5.05% (range 40–64). This frequency is significantly reduced compared with control values (P=0.001) (Table IV). Although the mean (±SD) chiasma frequency detected in the t(10;14) carrier (50.3±4.72%) was also lower than the observed control measures, the difference was not significant.

The incidence of sex chromosome univalents was 13.1% in the t(11;17) carrier and 28.8% in the t(10;14) carrier.

Segregation analysis on MII
A total of 144 MII spreads from the t(11;17) carrier and 36 MII spreads from the t(10;14) carrier were analysed. The presence of asymmetric dyads, i.e. chromosomes with both one normal and one derivative sister chromatid, were quantified. The number of asymmetric dyads expected to be found in MII can be calculated from the frequency of interstitial chiasmata in MI, and then compared with the frequency observed in MII. In the case of double interstitial chiasmata, half of them would result in no net exchange of the translocated segment. We did not find significant differences between the expected and the observed frequencies of asymmetric dyads in the two translocation carriers ({chi}2-test). In the t(10;14) carrier, the frequencies of dimorphic chromosomes detected in MII were 72.2% and 75.0% for chromosomes 10 and 14, respectively (Table V), and in the t(11;17) carrier, frequencies were 0.7% and 1.4% for chromosomes 11 and 17, respectively. Segregation modes were determined in both subjects. There was no evidence for 4:0 segregation. Regarding the t(11;17) carrier, alternate, adjacent 1, adjacent 2 and 3:1 segregation represented 47.2%, 23.6%, 24.3% and 2.8% of the analysed MII, respectively. A small proportion of MII (2.1%) presented asymmetric dyads, thus precluding the determination of the segregation mode (alternate or adjacent 1). In the t(10;14) carrier, the proportion of spreads with asymmetric dyads was 88.9%, of which 13.9% could be assigned to an adjacent 2 segregation (global frequency 16.7%) and 2.8% to a 3:1 segregation. A total of 72.2% could have originated from either alternate or adjacent 1 segregation. An additional 8.3% of the cells were products of alternate segregation. These results are summarized in Table VI.


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Table V. Frequencies of asymmetric dyads expected and found at MII for t(10;14) translocation carrier

 

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Table VI. Segregation mode frequencies found at MII and frequencies of normal/balanced (N/B) and unbalanced (U) forms for each type of segregation

 
FISH on decondensed sperm nuclei
A total of 5371 spermatozoa from the t(11;17) carrier and 3111 spermatozoa from the t(10;14) carrier were analysed by triple-colour FISH in order to perform the segregation analysis (Table VII). For the t(11;17) carrier and the t(10;14) carrier, the total number of balanced spermatozoa with a normal or balanced phenotype was found to be 40.8% and 44.6%, respectively. Adjacent 1 phenotypes were present in 26.1% of the analysed sperm nuclei from the t(11;17) carrier. The frequency of this segregation mode was 38.6% in the t(10;14) carrier. In both carriers, the observed proportions of the two reciprocal types of adjacent 1 products were significantly different. In the t(11;17) carrier, products containing the long translocated segment were found more frequently (P=0.0031), whereas in the t(10;14) carrier, products containing the short translocated segment had an increased incidence (P<0.0221).


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Table VII. Segregation mode frequencies found at sperm nuclei

 
All adjacent 2 phenotypes were observed in the two patients, including those resulting from recombination events within interstitial segments. As expected from the interstitial chiasma frequency data obtained in the MI analysis, the proportion of adjacent 2 recombinant phenotypes was higher in the t(10;14) carrier. Frequencies observed for the global amount of adjacent 2 phenotypes were 25.7% for the t(11;17) carrier and 11.9% for the t(10;14) carrier.

Finally, the eight different 3:1 phenotypes were detected in the two patients. Monosomic nuclei frequency was 4.4% in the t(11;17) carrier and 4.0% in the t(10;14) carrier, which represents a significant increase compared with the frequency of trisomic nuclei [3.0% in the t(11;17) carrier, 0.9% in the t(10;14) carrier].

In the aneuploidy assay, 10 137 spermatozoa from the t(11;17) carrier and 10 125 spermatozoa from the t(10;14) carrier were analysed for chromosomes 6 and 21, and 10 141 spermatozoa from the t(11;17) carrier and 10 044 from the t(10;14) carrier, for chromosomes X, Y and 18. A total of 20 107 and 20 560 spermatozoa from controls were recorded for chromosomes 6 and 21 and for chromosomes X, Y and 18, respectively. Results have been compared with control frequencies, and no significant variations have been found. These results are summarized in Table VIII.


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Table VIII. Aneuploidy frequencies detected in the two translocation carriers

 

    Discussion
 Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Acknowledgements
 References
 
Some articles have proposed the presence of sperm selection against specific chromosomal contents (Estop et al., 1999Go); however, our results do not support this hypothesis, as no significant differences have been found between the segregation frequencies at the MII and the segregation frequencies at the sperm level.

Chiasma analysis
Closed rings were the most common quadrivalent configuration found in the meiotic analysis in MI of the two translocation carriers. No univalents, bivalents or trivalents of the translocated chromosomes were detected. This is consistent with the behaviour observed in other reciprocal translocations (Templado et al., 1990Go; Goldman and Hultén, 1993aGo; Armstrong et al., 2000Go). It has been suggested that a short translocated segment could lead to the formation of open chain configurations, due to a failure of either in the pairing process or the chiasma formation in this region. However, this is not the case for the t(10;14), where the small size of the translocated segment of chromosome 14 has not been a limiting factor to produce a ring configuration during MI. Other authors have reported similar behaviour in translocation carriers with short exchanges (Chandley et al., 1976Go; Goldman and Hultén, 1993aGo; Armstrong et al., 2000Go).

When compared with data obtained from meiotically normal men, the mean chiasma frequency was decreased in the t(11;17) carrier, but not in the t(10;14) carrier. A reduction of chiasmata has also been reported in other translocation carriers (Goldman and Hultén, 1993aGo; bGo).

The frequency of chiasmata within the quadrivalent compared with control bivalents did not change for the chromosomes involved in t(11;17), but was significantly increased in the t(10;14) carrier, specifically in the long arms of chromosomes 10 and 14. Moreover, the distribution of chiasmata in both long arms was modified in such a way that the increase was mainly localized in the interstitial region. Other authors have reported modifications in chiasma frequency and distribution in translocation carriers (Chandley et al., 1976Go; Laurie et al., 1984Go; Goldman and Hultén, 1993aGo; bGo; Armstrong et al., 2000Go), and also in an insertion carrier (Goldman et al., 1992Go). However, the recombination behaviour within the context of a quadrivalent of a given chromosome is not predictable, since it has been seen that the same chromosome can display different chiasma patterns depending on the translocation. This is the case, for instance, in chromosome 10, which has been analysed in two other, different reorganizations (Laurie et al., 1984Go). Although the breakpoint for chromosome 10 is the same in these two translocations and in the translocation t(10;14) described in this work, the results obtained by Laurie et al. (1984)Go showed that the frequency of chiasma in 10q varies significantly among these three reorganizations. Since the frequency of interstitial chiasmata affects the final production of normal gametes, it is important to obtain information about the probability of this phenomenon, to predict the segregation pattern of reciprocal translocations.

Segregation analysis
As was expected from the different frequencies of interstitial chiasmata found in MI in the two subjects, the incidence of asymmetric dyads was specially high in the t(10;14) carrier and sporadic in the t(11;17) carrier. Assuming that half of the asymmetric dyads would result in a normal or balanced segregation, the data obtained from the t(11;17) carrier indicate that the proportion of balanced and unbalanced gametes produced by this heterozygote translocation carrier would be 48.3% and 52.7%, respectively. So, the genetic risk due to unbalanced gametes would be 52.7%. Data reported for the t(10;14) carrier lead to the prediction that the frequencies of balanced and unbalanced gametes would be 52.7% and 47.3%, respectively. In this case, the genetic risk would be 47.3%. These results are not significantly different from the segregation data obtained in decondensed sperm nuclei (Table VII). It can be assumed, then, that a selection based on chromosome contents does not occur during spermatogenesis. Similarly, in a recent meiotic study of a subject carrier of the most frequent reciprocal translocation in humans, t(11;22), the authors demonstrated that the high frequency of a 3:1 imbalance at birth observed in the offspring off carriers of t(11;22) does not imply a tendency towards the production of this type of segregation, but rather a strong postzygotic selection against other unbalanced genotypes (Armstrong et al., 2000Go).

Significant differences among complementary products of the adjacent 1 segregation mode were detected in sperm head analysis. This has also been observed in other reciprocal translocation analyses (Van Hummelen et al., 1997Go; Blanco et al., 1998Go). It has been proposed that the differences observed could be caused by unresolved chiasmata in MI, especially in the translocated segments, which would lead to either an arrest of the meiotic process or to the production of other abnormal phenotypes. In this case, the phenotypes expected to be found more frequently should have been those carrying the shorter translocated segments, i.e. 17,der(11) and 10,der(14). However, in the t(11;17) carrier the phenotypes with the longer translocated segment were seen with a significantly increased frequency. Other authors have reported a similar situation (Estop et al., 1998Go; Cifuentes et al., 1999Go; Honda et al., 1999Go; Oliver-Bonet et al., 2001Go). Thus, it is probable that other factors could modify the expected 1:1 ratio of reciprocal products. These factors could be related to cellular degeneration caused by the lack or the duplication of some essential spermatogenic genes located in one of the translocated segments, but also to hybridization failures of some of the fluorescent probes.

Aneuploidy analysis
Results obtained in the aneuploidy analysis performed on the two subjects did not show any evidence for an ICE (Table VIII). The two translocation carriers analysed in this study had normal semenograms. It has been suggested that the presence of ICE in translocation carriers could be restricted to those carriers with abnormal semenograms (Pellestor et al., 2001Go). Moreover, a correlation between poor semen parameters and an increase in the aneuploidy of other chromosomes not related to translocation has been detected in another work (Vegetti et al., 2000Go).

Centromeric probes for the sex chromosomes have been used on MI and MII spreads, allowing us to perform the behaviour analysis of the sex chromosomes during spermatogenesis. Surprisingly, the incidence of sex univalents in MI was extremely high: 13.1% in the t(11;17) carrier and 28.8% in the t(10;14) carrier. However, this incidence decreased progressively in subsequent spermatogenic stages. Thus, in MII, the number of cells bearing a sex chromosome disomy represented 0.09% in the t(11;17) carrier and 0.15% in the t(10;14) carrier, and in sperm nuclei the frequency decreased to reach the rates shown in Table VIII [0.38% in the t(11;17) carrier and 0.23% in the t(10;14) carrier]. A reduction of sex chromosome aneuploidy has also been observed in a meiotic study performed on a 47,XYY man (Blanco et al., 2001Go). In a study performed on azoospermic patients (Yogev et al., 2000Go), the authors concluded that the rate of the sex bivalent during meiosis was a good indicator for successful completion of spermatogenesis on these patients. It could be possible, then, that MI cells with unpaired sex chromosomes found in our analysis would experience a breakdown further on the meiotic process. In fact, it has been suggested that the presence of pairing impairment in one bivalent could be detected by a checkpoint control mechanism, thus leading to a complete failure of meiosis (Odorisio et al., 1998Go).

As mentioned above, selection against cellular chromosome content was not detected in these two patients. In conclusion, our results suggest that meiotic arrest processes in translocation carriers would be more conditioned by the presence of univalents rather than by chromosomal imbalances.


    Acknowledgements
 Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Acknowledgements
 References
 
We thank Àngels Niubó for technical assistance. We acknowledge the financial support given by FIS (PI020258) and CIRIT (2001 SGR-00201).


    References
 Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Acknowledgements
 References
 
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Submitted on October 2, 2003; resubmitted on June 14, 2004; accepted on August 2, 2004.