1 CNRS-UPR 1142, F-34396 Montpellier and 2 Cytogenetics Laboratory, C.H.U. Arnaud de Villeneuve, F-34033 Montpellier, France
![]() |
Abstract |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Key words: FISH/interchromosomal effect/PRINS/rearrangements/spermatozoa
![]() |
Introduction |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
|
![]() |
Materials and methods |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Group 1 (normal semen and abnormal karyotype)
Donor A, aged 34 years, carried a t(14;22) (q10;q10) Robertsonian translocation ascertained through systematic sperm donor screening for artificial insemination by donor. He has two normal children. His spermogram was normal with a sperm count of 77x106/ml and a motility of 70%. Donor B, aged 44 years, was heterozygous for a t(1;14) (p22;q21) ascertained after his wife underwent amniocentesis for advanced maternal age. He was the father of three normal children. His spermogram was normal with a sperm count of 85x106/ml and a motility of 60%. Donor C, aged 36 years, carried a t(7;9) (q33;p21) inherited through his mother. He was the father of a healthy 7 year old child. His spermogram was normal with a sperm count of 80x106/ml and a motility of 65%. Donor D, aged 30 years, was heterozygous for a t(7;18) (q35;q11) and was ascertained because of repeated reproductive failure. His wife had three spontaneous abortions. The spermogram of this man was normal with a sperm count of 43x106/ml and a motility of 55%. The study of meiotic segregation in spermatozoa of subjects C and D has been previously reported (Pellestor et al., 1997). Donor E, aged 35 years, carried a t(17;18) (p11;q11) inherited through his father. His spermogram was normal with a sperm count of 50x106/ml and a motility of 50%. Donor F, aged 38 years, carried an inv(2) (p11q13). The rearrangement was ascertained after two reproductive failures that ended in termination. The spermogram of this man was normal with a sperm count of 72x106/ml and a motility of 60%.
Group 2 abnormal semen and abnormal karyotype)
Donor G, aged 39 years, was heterozygous for a t(8;13) (p22;q13) ascertained after 4 years of infertility. Semen analysis revealed an oligoasthenoteratozoospermia (semen count 4.3x106/ml, motility 13%, nomal morphology 10%). Donor H, 30 years old, carried a t(13;15) (q10;q10) Robertsonian translocation, ascertained through a fertility work-up after 3 years of infertility. Seminal parameters showed oligoasthenoteratozoospermia (semen count 3.2x106/ml, motility 20%, normal morphology 8%). Donor I, aged 35 years, carried a t(5;9) (p12;p11) inherited through his mother. His sister was also carrier of the same translocation and had two spontaneous abortions. The semen analysis of this subject revealed an oligoteratozoospermia (semen count 15x106/ml, motility 40%, normal morphology 11%).
Controls
Three healthy normal volunteers, aged 27, 32 and 37 years, provided sperm specimens during the same period as the rearrangement carriers, and constituted the control group (group 3). The three subjects were of proven fertility and had normal seminal parameters (sperm count >50x106/ml, motility >50%, normal morphology >20%).
Sperm treatment
All experiments were performed on fresh ejaculates. The processing of sperm samples was identical for carrier and control specimens. The ejaculate was collected in a sterile container and kept at room temperature for 30 min. After liquefaction, an aliquot of the specimen was used for semen analysis. The rest of the sample was washed twice in phosphate-buffered saline (PBS) by centrifugation (8 min at 300 g) and fixed for 1 h in fresh fixative (3:1 methanol:glacial acetic acid) at 20°C. The sperm suspension was then dropped onto clean microscope slides and air-dried. Slides were aged 3 days at room temperature before use for in-situ chromosomal labelling. Before PRINS or FISH procedure, the slides were immersed in a 3 mol/l NaOH solution at room temperature for 5 min, passed through ethanol series (70, 90, 100%) and air-dried. The use of 3 mol/l NaOH solution allowed the simultaneous decondensation and denaturation of sperm nuclei, enabling rapid control of the degree of nucleus decondensation under the microscope.
In-situ chromosomal labelling procedures
For each subject, the same combination of probes or primers was used. Details are given in Table II.
|
PRINS labelling
Specific oligonucleotide primers for satellite DNA of chromosomes 1, 9, 13, 16, 21, X and Y were used in double or triple PRINS reactions. The specificity and the efficiency of these primers were previously tested in both metaphase and interphase nuclei. Their sequences and optimal technical conditions have been given elsewhere (Coullin et al., 1997). PRINS reactions were performed according to our sequential PRINS protocol (Pellestor et al., 1996
). For each primer, a mix was prepared in a final volume of 50 µl containing the oligonucleotide (50-250 pmol/l), the nucleotide mixture including a labelled dUTP (fluorescein-12-dUTP, rhodamine-4-dUTP or coumarin-6-dUTP), the Taq polymerase buffer and 2 units of Taq DNA polymerase (Roche Diagnostics). The mix was preheated on a waterbath at the annealing temperature of the used primer. The denatured preparation slides were put on the plate block of the thermocycler. The reaction consisted of two programmed steps: 10 min at the specific annealing temperature of the primer and 1030 min at 72°C in order to allow the nucleotide chain elongation. The slides were not sealed. Both the volume of the mix and the short incubation time prevented the slides from drying during the reaction. The PRINS reaction was stopped by immersing the slides in a stop buffer (500 mmol/l NaCl, 50 mmol/l EDTA, pH 8) at 72°C for 1 min, and the slides were transferred to 2xSSC, 0.5% Tween 20 at room temperature. The slides were then treated for 15 min at 37°C with a dideoxynucleotide mix and 2 units of Klenow enzyme in order to block the free 3'-ends of the elongation fragments generated by the first PRINS reaction. This intermediate step prevented mixing of labelling. The slides were washed and again placed on the plate block of the thermocycler. A second and then a third PRINS reaction mix, involving a primer specific for another chromosome and another labelled dUTP, was applied to the slides and a new reaction was run. Finally, the preparations were washed twice in 2xSSC, 0.05% Tween 20 at 37°C and counterstained with DAPI (0.5 µg/ml) and propidium iodide (0.02 µg/ml) in antifade solution (Vectashield).
Microscopy
Slides were viewed under a Leitz fluorescence microscope DRMB (Leica SA, Rueil-Malmaison, France), equipped with a DAPI single band-pass filter, a fluorescein single band-pass filter, a rhodamine single band-pass filter, a fluorescein/rhodamine double band-pass filter, and a triple filter set for simultaneous observation of fluorescein, rhodamine and coumarin signals.
Sperm scoring
Slides were analysed by two independent observers, each scoring ~5000 nuclei per slide. Scorers were blinded with respect to the identity of the specimen. The scored sperm nuclei were intact, not overlapping and with a well-defined border. Nuclei that were decondensed to greater than thrice the size of an undecondensed spermatozoa were not scored because of the diffusion of fluorescent signals. The haploid nuclei displayed two spots of different colour corresponding to the two labelled chromosomes. Nuclei were scored as disomic when they displayed a total of three signals. Two of these signals had to be similar in size, colour and intensity, and separated from each other by at least one signal domain diameter. Nuclei were classified as diploid when they contained four distinct fluorescent spots, two for each tested chromosome. We scored for disomy and diploidy but not for nullisomy since failure to detect a signal could be the result of technical artefact as well as non-disjunction.
Statistical analyses
Student's t-test was used to verify the homogeneity of mean ages in the three groups of subjects. The 2-test was used: (i) to check the homogeneity of disomy rates for the 10 chromosomes tested in each sperm sample (interchromosomal variation); (ii) to compare the frequencies of disomy for each chromosome between the 12 subjects (interindividual variation); (iii) to test the variability of the diploidy frequencies. A value of P < 0.05 was considered to be significant. The homogeneity of the mean autosomal disomy rates among groups was also tested by the non-parametric Mann-Whitney U-test. P < 0.05 was considered to be significant.
![]() |
Results |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The labelling efficiency of each probe and primer was determined by scoring the proportion of labelled nuclei on samples of 1000 sperm nuclei. The labelling efficiency was similar in FISH and PRINS assays (from 98.4 to 99.9% of labelled nuclei according to both the sperm samples and the chromosomes analysed), but we noted a slightly higher efficiency in samples from normal men compared to rearrangement carriers (mean values 99.5 versus 98.7%). For all chromosomes tested, no significant difference (P > 0.05) was found between the results of FISH and PRINS assays. The details of disomy rates per subject and chromosome are given in Table III.
|
Interchromosomal variation
Variations of disomy rates were first estimated separately in autosomes and gonosomes. In the pooled analysis, we considered only the total gonosome disomy rate (XX+ XY+YY) for statistical analysis. Results are given in Table IV.
|
In group 2, the three rearrangements displayed significant interchromosomal variations (P < 0.05) in autosomes as well as in pooled data (autosomes + gonosomes) (P < 0.01). No significant variation (P > 0.05) was found for disomy in gonosomes.
In the control group, only donor C1 showed significant interchromosomal variations in autosomes (2 = 17.39; P < 0.02). Variations were significant (P < 0.01) in the three donors when autosome and gonosome data were pooled.
Details of the calculation indicated that in both the translocation t(7;9) and the inversion inv(2), the significance was reached because of the high incidence of disomy scored for chromosome 21. In fact, chromosome 21 consistently displayed a higher disomy frequency (from 0.22 to 0.55%) than the other autosomes tested in both rearrangement carriers and control donors (Table III).
Interindividual variation
In group 1, the disomy values found for each chromosome were homogeneous (P > 0.10), except for chromosome 15 (2 = 11.37; P < 0.05) and chromosome X (
2 = 12.93; P < 0.05) (Table V
). In group 2, there were significant interdonor differences in the disomy frequency for chromosome 1 (
2 = 8.41; P < 0.02), chromosome 15 (
2 = 6.70; P < 0.01) and chromosome Y (
2 = 10.75; P < 0.01). The control group displayed interindividual variations only for chromosome 1 (
2 = 6.29; P < 0.05).
|
|
|
![]() |
Discussion |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
All the control values are within the limits of the disomy rates usually reported for normal subjects (Guttenbach et al., 1997). Both interchromosomal and interindividual variations displayed different patterns according to the semen quality. In the control group and patients with normal semen parameters (group 1), interchromosomal variations were limited in autosomes and non-existant in gonosomes. Variations were highly significant in patients with abnormal sperm (group 2), or globally when autosome and gonosome data were pooled. Chromosome analysis of spermatozoa from infertile males has shown that these patients present a high variability in the behaviour of chromosome segregation during meiosis (Miharu et al., 1994
; Moosani et al., 1995
). This variability may be linked to the multifactorial nature of male infertility, but in cases of subjects combining poor semen quality and chromosomal rearrangement, interchromosomal variations in disomy rates may directly reflect meiotic impairment. It might be suggested that both fluctuation in disomy and degree of semen parameter abnormalities are conditioned by the chromosomes involved in the rearrangement.
The analysis of our results indicates that the sex chromosomes and chromosome 21 have a higher frequency of disomy than the other chromosomes tested. This finding confirms previous reported data (Spriggs et al., 1995; Blanco et al., 1996
; Pellestor et al., 1996
). Increase of disomy for chromosome 21 and sex chromosomes was not specific to rearrangement carriers but observed in all subjects tested. This observation disagrees with the hypothesis of a relationship between the increase of non-disjunction for these chromosomes and the presence of a chromosomal rearrangement, but rather is consistent with the suggestion that sex chromosomes and chromosome 21 might be more susceptible to non-disjunction than other chromosomes. Data from spontaneous abortions and livebirths support this idea (Jacobs, 1992
).
Interindividual variations in disomy rate are restricted to a few chromosomes in each group of subjects (Table V). The significant variations concern chromosomes 1, 15 and sex chromosomes. Interdonor variations for the same chromosome have been reported (Spriggs et al., 1995
, 1996
). These findings are too limited to drawn conclusions on interindividual fluctuations of non-disjunction events. However, molecular approaches offer the possibility to investigate this question, in particular with regard to the recent data on the importance of interindividual difference in the rate of total genomic recombination and genetic control of crossover frequency (Brown et al., 2000
).
According to our finding, rearrangement carriers with normal semen parameters displayed no significant difference in disomy rates with the normal subjects, except for chromosome 4. The homogeneity of disomy rate between the two groups was confirmed by two statistical tests (Tables V and VI). This result provided no direct evidence for the occurrence of an interchromosomal effect in sperm of fertile rearrangement carriers. On the contrary, infertile men carrying a translocation (group 2) showed significant interindividual variation when compared to control donors. All chromosomes tested, except chromosome 16, displayed a significant difference in disomy rate, and significant variations were also found for several chromosomes between groups 1 and 2. These results support the hypothesis of an interchromosomal effect restricted to the cases of translocation carriers with abnormal semen parameters. This is consistent with a study (Vegetti et al., 2000
) which found a higher incidence of diploidy and aneuploidy in sperm of infertile men with abnormal karyotypes compared to normal controls. The analysis of several chromosomes belonging to different chromosomal groups does not support the hypothesis of an effect restricted to a particular chromosome group. However, this result needs to be confirmed by similar analysis on other chromosomes. Several studies have reported a correlation between poor semen quality and increased frequency of aneuploidy in spermatozoa (Moosani et al., 1995
; Rives et al., 1999
). In cases of rearrangement carriers, such a correlation may depend on the chromosomes involved in the rearrangement and their breakpoints. Pachytene configurations of some structural rearrangements may more directly affect the progression of meiosis and the production of gametes when they are closely associated with the sex vesicle or other autosomal bivalent (Guichaoua et al., 1991
; Navarro et al., 1991
). The length of the translocated segments could be an essential factor in the occurrence of an association with another bivalent, and facilitate the malsegregation of other chromosome pairs (Blanco et al., 2000
). These associations may lead to synaptic anomaly in various chromosome pairs and spermatogenesis impairment (Luciani et al., 1984
; Batanian and Hultén, 1987
). The influence of defective pairing of some chromosomes on the segregation of other chromosomes has been evidenced directly in human trisomic oocytes (Cheng et al., 1998
) and XO mice oocytes (Hunt et al., 1995
). However, it seems that the female meiotic checkpoint process was less efficient than the male mechanism (LeMaire-Adkins et al., 1997
). Disturbances in the pairing process can adversely block the male meiotic progression, which can limit the occurrence of an interchromosomal effect in mature spermatozoa. An interesting observation is also the higher frequency of diploidy found in spermatozoa from infertile rearrangement carriers, which can also be related to synaptic abnormalities. Elevated frequency of diploidy in spermatozoa of rearrangement carriers has been reported in several studies (Table I
) and could also reflect a gross disruption of meiosis with the absence of cytokinesis. If both timing accuracy of chromosome pairing and recombination act as checkpoints in meiosis, the presence of abnormal pairing configurations could upset the progression through meiosis and lead to the increased production of diploid cells (Goldman and Hultén, 1993
; Kleckner, 1996
).
This interchromosomal effect may be restricted to males since the sterilizing effect of translocations appears to be limited to male heterozygotes. No effects on germ-cell development have been found in women carrying translocations.When compared to the high maternal contribution to chromosomal abnormalities of the human conceptus (Pellestor, 1991; Nakaoka et al., 1998
), the increase of disomy in spermatozoa of infertile carriers of rearrangements may be of limited clinical releveance, except for sex chromosomes where male non-disjunction is in the same order of magnitude as in female meiosis. However, these data need to be taken into account for counselling of couples with a male partner carrying a translocation before treatment of infertility by intracytoplasmic sperm injection (ICSI), since a higher incidence of sex chromosomal aneuploidy of paternal origin has been reported in children conceived by ICSI (In't Veld et al., 1995).
Most cases of interchromosomal effect reported concerned fortuitous observations of trisomy 21 in the lineage of balanced rearrangement carriers. As pointed out elsewhere (Lindenbaum et al., 1985), a significant prevalence of rearrangements in parents of trisomy 21 offspring does not necessarily imply that these rearrangements increase the likehood of parental meiotic non-disjunction. In the case of fertile male rearrangement carriers, it has been demonstrated (Schinzel et al., 1992
) that such associations are fortuitous, since the concomittant non-disjunctions originated in the mother. The potential high incidence of balanced rearrangements in the parents of trisomic 21 offspring could largely result from the higher age of parents due to the difficulty which carriers of chromosomal rearrangements and their partners have to conceive and to carry a pregnancy to term. In fact, the risk of association between a balanced chromosomal rearrangement and trisomy 21 in fertile couples may be assumed to be very low and to correspond to the risk for trisomy 21 in a normal population.
Various lines of evidence do not support the concept of a paternal interchromosomal effect for carriers of structural chromosomal aberrations. Our data indicate that the occurrence of interchromosomal effect may be restricted to the infertile rearrangement carriers. We can assume that the choice of the rearrangements investigated, the use of two different labelling techniques and the number of chromosomes analysed confers a good reliability to the present results. The only restrictive parameter which was not possible to control was the large diversity of structural rearrangements in humans. This is particularly true in reciprocal translocations since almost all observed translocations are unique in terms of configuration and meiotic behaviour. To date, sperm analysis has revealed that the frequecy of chromosomally unbalanced sperm related to translocation varied dramatically from 0 to 77% (Martin and Spriggs, 1995; Pellestor et al., 1997
). The production of unbalanced gametes depends on various criteria involving the position of breakpoints, the size of the translocated segments and the genetic background. All these factors may affect the occurrence of an interchromosomal effect. Consequently, the present findings need to be confirmed by the sperm analysis of other rearrangements. Nevertheless, the association of poor semen parameters and increased aneuploidy in conjunction with a structural rearrangement probably reflects important meiotic disturbances and should consequently be considered when counselling males heterozygous for a chromosomal rearrangement.
![]() |
Acknowledgements |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
![]() |
Notes |
---|
![]() |
References |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Blanco, J., Egozcue, J. and Vidal, F. (1996) Incidence of chromosome 21 disomy in human spermatozoa as determined by fluorescent in-situ hybridization. Hum. Reprod., 11, 722726.[Abstract]
Blanco, J., Egozcue, J., Clusellas, N. et al. (1998) FISH on sperm heads allows the analysis of chromosome segregation and interchromosomal effects in carriers of structural rearrangements: results in a translocation carrier, t(5;8) (q33;q13). Cytogenet. Cell Genet., 83, 275280.[ISI][Medline]
Blanco, J., Egozcue, J. and Vidal, F. (2000) Interchromosomal effects for chromosome 21 in carriers of structural chromosome reorganizations determined by fluorescence in situ hybridization on sperm nuclei. Hum. Genet., 106, 500505.[ISI][Medline]
Brown, A.S., Feingold, E., Broman, K.W. et al. (2000) Genome-wide variation in recombination in female meiosis: a risk factor for non-disjunction of chromosome 21. Hum. Mol. Genet., 9, 515523.
Burns, J.P., Koduru, P.R.K., Alonso, M.L. et al. (1986) Analysis of meiotic segregation in a man heterozygous for two reciprocal translocations using the humster in vitro penetration system. Am. J. Hum. Genet., 38, 954964.[ISI][Medline]
Canki, N. and Dutrillaux, B. (1979) Two cases of familial paracentric inversion in man associated with sex chromosome anomaly 47,XXY,inv(5) (q21q32) and 45,X,inv(7) (q11.3q22.3). Hum. Genet., 47, 261265.
Cheng, E.Y., Chen, Y.J., Bonnet, G. et al. (1998) An analysis of meiotic pairing in trisomy 21 oocytes using fluorescent in situ hybridization. Cytogenet. Cell Genet., 80, 4853.[ISI][Medline]
Cifuentes, P., Navarro, J., Blanco, J. et al. (1999) Cytogenetic analysis of sperm chromosomes and sperm nuclei in a male heterozygous for a reciprocal translocation t(5;7) (q21;q32) by in situ hybridization. Eur. J. Hum. Genet., 7, 231238.[ISI][Medline]
Colls, P., Blanco, J., Martinez-Pasarell, O. et al. (1997) Chromosome segregation in a man heterozygous for a pericentric inversion, inv(9) (p11q13), analyzed by using sperm karyotyping and two-color fluorescence in situ hybridization on sperm nuclei. Hum. Genet., 99, 761765.[ISI][Medline]
Coullin, P., Andréo, B., Charlieu, J.P. et al. (1997) Primed in situ (PRINS) labeling with Alu and satellite primers for rapid characterization of human chromosomes in hybrid cell lines. Chromosome Res., 5, 307312.[ISI][Medline]
Couzin, D.A., Watt, J.L. and Stephen, G.S. (1987) Structural rearrangements in the parents of children with primary trisomy 21. J. Med. Genet., 24, 280282.[Abstract]
Estop, A.M., Cieply, K., Munné, S. et al. (2000) Is there an interchromosomal effect in reciprocal translocation carriers? Sperm FISH studies. Hum. Genet., 106, 517524.
Giltay, J.C., Kastrop, P.M.M., Tiemessen, C.H.J. et al. (1999) Sperm analysis in a subfertile male with a Y;16 translocation, using four color FISH. Cytogenet. Cell Genet., 84, 6772.[ISI][Medline]
Goldman, A.S.H. and Hultén, M.A. (1993) Meiotic analysis by FISH of a human male 46,XY,t(15;20) (q11.2;q11.2) translocation heterozygote: quadrivalent configuration, orientation and first meiotic segregation. Chromosoma, 102, 102111.
Grell, R.F. (1962) A new hypothesis on the nature and sequence of meiotic events in the female of Drosophila melanogaster. Proc. Natl. Acad. Sci. USA, 48, 165172.[ISI][Medline]
Guichaoua, M.R., de Lanversin, A., Cataldo, C. et al. (1991) Three dimensional reconstruction of human pachytene spermatocyte nuclei of a 17;21 reciprocal translocation carrier: study of XY-autosome relationships. Hum. Genet., 87, 709715.[ISI][Medline]
Guttenbach, M. and Schmid, M. (1990) Determination of Y chromosome aneuploidy in human sperm nuclei by nonradioactive in situ hybridization. Am. J. Hum. Genet., 46, 553558.[ISI][Medline]
Guttenbach, M., Engel, W. and Schmid, M. (1997) Analysis of structural and numerical chromosome abnormalities in sperm of normal men and carriers of constitutional chromosome aberrations. A review. Hum. Genet., 100, 121.[ISI][Medline]
Hecht, F. and Patil, S.R. (1977) Interchromosomal effect in man. Clin. Genet., 12, 189190.[ISI]
Honda, H., Miharu, N., Ohashi, Y. et al. (1999) Analysis of segregation and aneuploidy in two reciprocal translocation carriers, t(3;9) (q26.2;q32) and t(3;9) (p25;q32), by triple-color fluorescence in situ hybridization. Hum. Genet., 105, 428436.
Hunt, P., LeMaire, R., Embury, P. et al. (1995) Analysis of chromosome behavior in intact mammalian oocytes: monitoring the segregation of a univalent chromosome during female meiosis. Hum. Mol. Genet., 4, 20072012.[Abstract]
In't Veld, P.A., Brandenburg, H., Verhoeff, A. et al. (1995) Sex chromosomal abnormalities and intracytoplasmic sperm injection. Lancet, 346, 773.[ISI][Medline]
Jacobs, P.A. (1992) The chromosome complement of human gametes. Oxford Rev. Reprod. Biol., 12, 4872.
LeMaire-Adkins, R., Radke, K. and Hunt, P.A. (1997) Lack of checkpoint control at the metaphase/anaphase transition: a mechanism of meiotic nondisjunction in mammalian females. J. Cell Biol., 139, 16111619.
Lejeune, J. (1963) Autosomal disorders. Pediatrics, 32, 326337.[Abstract]
Kleckner, I. (1996) Meiosis: how could it work ? Proc. Natl Acad. Sci. USA, 93, 81678174.
Lindenbaum, R.H., Hultén, M., McDermott, A. et al. (1985) The prevalence of translocations in parents of children with regular trisomy 21: a possible interchromosomal effect? J. Med. Genet., 22, 2428.[Abstract]
Lu, P.Y., Hammitt, D.G., Zinsmeister, A.R. et al. (1994) Dual color fluorescence in situ hybridization to investigate aneuploidy in sperm from 33 normal males and a man with a t(2;4;8) (q23;q27;p21). Fertil. Steril., 62, 394399.[ISI][Medline]
Luciani, J.M., Guichaoua, M.R., Mattei, A. et al. (1984) Pachytene analysis of a man with a 13q;14q translocation and infertility. Cytogenet. Cell Genet., 38, 1422.[ISI][Medline]
Martin, R.H. and Spriggs, E.L. (1995) Sperm chromosome complements in a man heterozygous for a reciprocal translocation 46, XY,t(9;13) (q21.1;q21.2) and a review of the literature. Clin. Genet., 47, 4246.
Martini, E., von Bergh, A.R.M., Coonen, E. et al. (1998) Detection of structural abnormalities in spermatozoa of a translocation carrier t(3;11) (q27.3;q24.3) by triple FISH. Hum. Genet., 102, 157165.
Mercier, S., Morel, F., Fellman, F. et al. (1998) Molecular analysis of the chromosomal equipment in spermatozoa of a 46,XY,t(7;8) (q11.21;cen) carrier by using fluorescence in situ hybridization. Hum. Genet., 102, 446451.
Miharu, N., Best, R.G. and Young, S.R. (1994) Numerical chromosome abnormalities in spermatozoa of fertile and infertile men detected by fluorescence in situ hybridization. Hum. Genet., 93, 502506.[ISI][Medline]
Moosani, N., Pattinson, H.A., Carter, M.D. et al. (1995) Chromosomal analysis of sperm from men with idiopathic infertility using sperm karyotyping and fluorescence in situ hybridization. Fertil. Steril., 64, 811817.[ISI][Medline]
Nakaoka, Y., Okamoto, E., Miharu, N. et al. (1998) Chromosome analysis in human oocytes remaining unfertilized after in-vitro insemination: effect of maternal age and fertilization rate. Hum. Reprod., 13, 419424.[ISI][Medline]
Navarro, J., Vidal, F., Benet, J. et al. (1991) XY-trivalent association and synaptic anomalies in a male carrier of a Robertsonian t(13;14) translocation. Hum. Reprod., 6, 376381.[Abstract]
Pellestor, F. (1991) Differential distribution of aneuploidy in human gametes according to their sex. Hum. Reprod., 6, 12521258.[Abstract]
Pellestor, F., Girardet, A., Coignet, L. et al. (1996) Assessment of aneuploidy for chromosomes 8, 9, 13, 16 and 21 in human sperm by using Primed In Situ labeling technique. Am. J. Hum. Genet., 58, 797802.[ISI][Medline]
Pellestor, F., Girardet, A., Coignet, L. et al. (1997) Cytogenetic analysis of meiotic segregation in sperm from two males heterozygous for reciprocal translocations using PRINS and humster techniques. Cytogenet. Cell Genet., 78, 202208.[ISI][Medline]
Portin, P. and Rantanen, M. (1990) Further studies on the interchromosomal effect on crossing over in Drosophila melanogaster affecting the preconditions of exchange. Genetica, 82, 203-207.[ISI][Medline]
Rives, N., Saint Clair, A., Mazurier, S. et al. (1999) Relationship between clinical phenotype, semen parameters and aneuploidy frequency in sperm nuclei of infertile male. Hum. Genet., 105, 266272.[ISI][Medline]
Robbins, W.A., Segraves, R., Pinkel, D. et al. (1993) Detection of aneuploid human sperm fluorescence in situ hybridization: evidence for a donor difference in frequency of sperm disomic for chromosomes 1 and Y. Am. J. Hum. Genet., 52, 799807.[ISI][Medline]
Rousseaux, S., Chevret, E., Monteil, M. et al. (1995a) Sperm nuclei analysis of a Robertsonian t(14q21q) carrier by FISH using three plasmids and two YAC probes. Hum. Genet., 96, 655660.[ISI][Medline]
Rousseaux, S., Chevret, E., Monteil, M. et al. (1995b) Meiotic segregation in males heterozygote for reciprocal translocations: analysis of sperm nuclei by two and three colour fluorescence in situ hybridization. Cytogenet. Cell Genet., 71, 240246.[ISI][Medline]
Rudak, E., Jacobs, P.A. and Yanagimachi, R. (1978) Direct analysis of the chromosome constitution of human spermatozoa. Nature, 274, 911913.[ISI][Medline]
Schinzel, A.A., Adelsberger, P.A., Binkert, F. et al. (1992) No evidence for a paternal interchromosomal effect from analysis of the origin of nondisjunction in Down syndrome patients with concomitant familial chromosome rearrangements. Am. J. Hum. Genet., 50, 288293.[ISI][Medline]
Serra, A., Brahe, A., Millington-Ward, G. et al. (1990) Pericentric inversion of chromosome 9: prevalence in 300 Down syndrome families and molecular studies of nondisjunction. Am. J. Med. Genet., 7, 162168.
Spriggs, E.L., Rademaker, A.W. and Martin, R.H. (1995) Aneuploidy in human sperm: results of two and three color fluorescence in situ hybridization using centromeric probes for chromosomes 1, 12, 15, 18, X and Y. Cytogenet. Cell Genet., 71, 4753.[ISI][Medline]
Spriggs, E.L., Rademaker, A.W. and Martin, R.H. (1996) Aneuploidy in human sperm: the use of multicolor FISH to test various theories of nondisjunction. Am. J. Hum. Genet., 58, 356362.[ISI][Medline]
Stoll, C., Fiori, E. and Beshara, D. (1978) Interchromosomal effect in balanced translocations. Birth Defects, 14, 393398.
Van Hummelen, P., Manchester, D., Lowe, X.R. et al. (1997) Meiotic segregation, recombination and gamete aneuploidy assessed in a t(1;10) (p22.1;q22.3) reciprocal translocation carrier by three and four probe multicolor FISH in sperm. Am. J. Hum. Genet., 61, 651659.
Vegetti, W., Van Assche, E., Frias, A. et al. (2000) Correlation between semen parameters and sperm aneuploidy rates investigated by fluorescence in-situ hybridization in infertile men. Hum. Reprod., 15, 351365.
Submitted on September 4, 2000; accepted on February 12, 2001.