Hum. Reprod. Advance Access originally published online on November 6, 2007
Human Reproduction 2008 23(1):227-230; doi:10.1093/humrep/dem359
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Segregation of chromosomes in sperm of a t(X;18)(q11;p11.1) carrier inherited from his mother: Case Report
1 Laboratoire d’Histologie, Embryologie et Cytogénétique, Faculté de Médecine et des Sciences de la Santé, Université de Bretagne Occidentale, 22, Avenue Camille Desmoulins, CS 93837, F-29238 Brest Cedex 3, France 2 Service de Cytogénétique,Cytologie et Biologie de la Reproduction, CHU Morvan, Brest, France 3 Laboratoire de Kerlann, Vannes, France 4 Service de Biologie et Médecine du Développement et de la Reproduction, CHU Nantes, Nantes, France
5 Correspondence address. Tel: +33-2-98-01-64-76; Fax: +33-2-98-01-81-89; E-mail: marc.debraekeleer{at}univ-brest.fr
 |
Abstract |
|---|
 Top Abstract IntroductionCase ReportResultsDiscussionFundingReferences |
|---|
Balanced reciprocal translocations are the most common structural abnormalities; most involve two autosomes while a few involve a gonosome (X or Y chromosome) and an autosome. These rearrangements are usually associated with infertility and/or a higher risk of chromosomal imbalances among offspring. This 26 years old man was first seen because of a 3-year history of primary infertility. He had been found to have a translocation, t(X;18)(q11;p11.1), inherited from his mother when he was 9 years old. Semen analysis showed a very severe oligoasthenoteratozoospermia (OAT). A total of 447 spermatozoa were analysed using three-colour fluorescent in situ hybridization (FISH). The alternate segregation pattern, leading to a normal or balanced chromosomal content, was found in 54.36% of the spermatozoa studied. The frequencies of Adjacent I, Adjacent II, 3:1 segregation and diploidy (or 4:0 segregation) were 8.28, 5.14, 22.37 and 2.01%, respectively. Balanced reciprocal translocations between an autosome and the X chromosome lead to important disruptions in human spermatogenesis. Almost all the males with an X-autosome translocation have azoospermia. The man reported here had very severe OAT and is the first in whom the meiotic segregation pattern was analysed. This case further emphasizes the interest in performing FISH studies in infertile males with a chromosomal translocation to provide them with a personalized imbalance risk.
Key words: meiotic segregation/FISH/X-autosome translocation
|
Introduction |
|---|
|
TopAbstractIntroductionCase ReportResultsDiscussionFundingReferences |
|---|
Balanced reciprocal translocations are the most frequent structural chromosomal abnormalities in humans. Their frequency is estimated at 0.12% in the general population but is 6.5 times more frequent among infertile males (De Braekeleer and Dao, 1991
; De Braekeleer et al., 2006). They result from material exchange between two non-homologous chromosomes, without loss or gain of material. The majority of reciprocal translocations occurs between two autosomes. Translocations involving gonosomes are rare and belong to three subgroups: Y-autosome translocations (Hsu, 1994; Giltay et al., 1999; Brisset et al., 2005; Pinho et al., 2005), X-autosome translocations (Fraccaro et al., 1977; Lee et al., 2003; Ma et al., 2003; Ishikawa et al., 2007) and X–Y translocations (Yamada et al., 1982; Gabriel-Robez et al., 1990; Taiar et al., 1995; Morel et al., 2001). Most of the men carrying a translocation involving gonosomes have azoospermia (Lee et al., 2003; Brisset et al., 2005; Ishikawa et al., 2007), although some show a severe oligozoospermia (Fraccaro et al. 1977; Giltay et al. 1999; Morel et al. 2001; Ma et al., 2003).
Meiotic segregation, studied by heterospecific fecundation or fluorescent in situ hybridization (FISH), of >80 men carrying a translocation between two autosomes has been reported (Morel et al., 2006). However, there are only two reports on males having a Y-autosome translocation (Mennicke et al., 1997; Giltay et al., 1999) and a sole report concerning a t(X;Y) (Morel et al., 2001). To our knowledge, we report here the first meiotic segregation analysis in a male carrying an X-autosome translocation.
|
Case Report |
|---|
|
TopAbstractIntroductionCase ReportResultsDiscussionFundingReferences |
|---|
Patient
The patient was first seen when he was 9 years old, because of his parents’ secondary infertility following his birth. The cytogenetic analysis revealed a 46,Y,t(X;18)(q11;p11.1) karyotype, inherited from his mother.
The couple (female 24 years old; male 26 years old) presented with a 3-year history of primary infertility. The female partner had a normal karyotype. Semen analysis showed a very severe oligoasthenoteratozoospermia with 0.4 million spermatozoa/ml, 95% immobile sperm and 90% abnormal forms. Prior to this study, the patient was informed of the investigations and subsequently gave his consent.
Meiotic segregation analysis
Triple FISH was carried out using the specific alphoid probe of chromosome 18 (D18Z1, spectrum aqua, Abbott, Rungis, France), the 18p subtelomere probe (tel 18p, spectrum green, Abbott) and the subtelomere Xq/Yq probe (telXq/Yq, spectrum orange, Abbott). An ideogram showing the translocation, the localization of the probes and the quadrivalent is shown in Fig. 1.
|
The sperm sample of the patient was also analysed in triple FISH with specific alphoid probes of chromosomes Y (DYZ3, spectrum orange, Abbott) and 18 (D18Z1, spectrum aqua, Abbott) and the 18p subtelomere probe (tel 18p, spectrum green, Abbott).
Detailed procedures for sperm preparation and FISH have been previously described (Morel et al., 2004b; Douet-Guilbert et al., 2005; Morel et al., 2007). The slides were analysed using a Zeiss Axio Plan microscope (Zeiss, Le Pecq, France). Subsequent image acquisition was performed using a CCD camera with Isis (significant in situ imaging system) (MetaSystems, Altlussheim, Germany).
Spermatozoa with one green, one orange and one blue signal were classified as normal or balanced whereas spermatozoa with other signal combinations were scored as unbalanced.
|
Results |
|---|
|
TopAbstractIntroductionCase ReportResultsDiscussionFundingReferences |
|---|
A total of 447 spermatozoa was analysed (Table I). A preferential alternate segregation mode was observed with a rate of 54.36%. The probe combination could not distinguish whether the studied spermatozoa had a chromosomally balanced or normal content.
|
All other spermatozoa were unbalanced (45.64%). Among the unbalanced spermatozoa, the 3:1 segregation mode was the most frequent with 22.37% of analysed spermatozoa, followed by the Adjacent I (8.28%) and Adjacent II modes (5.14%). Moreover, 2.01% of the spermatozoa were diploid or segregated in the 4:0 mode. The remaining spermatozoa (7.83%) showed ambiguous signals or hybridization failure (Caer et al., 2007
). A total of 287 spermatozoa was analysed in triple FISH using DYZ3, D18Z1 and tel18p probes (Table II). The frequency of spermatozoa exhibiting one orange spot, one blue spot and one green spot was assessed at 29.27%. We could not distinguish normal (23,Y) from unbalanced (24,Y,der(X)) spermatozoa as they showed the same fluorescent combination. Nevertheless, with triple FISH using D18Z1, tel18p and telXqYq, we had observed that 2.68% of spermatozoa were 24,Y,der(X) (Table I). Thus, using this correction factor, we estimated that 26.59% (29.27–2.68) was the rate of normal spermatozoa (23,Y).
|
|
Discussion |
|---|
|
TopAbstractIntroductionCase ReportResultsDiscussionFundingReferences |
|---|
Balanced reciprocal translocations between two autosomes are the most common structural chromosomal rearrangements in humans. X-autosome translocations are rare and usually of maternal origin or arising de novo (Kalz-Fuller et al., 1999
). In general, female carriers exhibit normal reproductive function (Cantu et al., 1985; Ma et al., 2003; Panasiuk et al., 2004) while male carriers have azoospermia (Quack et al., 1988; Solari et al., 2001; Lee et al., 2003; Ishikawa et al., 2007).
Histologic examinations of biopsied testicular tissue from males carrying a X-autosome translocation show spermatogenetic arrest at the primary spermatocyte level with most of the cells being arrested at the pachytene stage (Quack et al., 1988), although a few germ cells at the spermatid level are sometimes found (Ishikawa et al., 2007). Indeed, balanced reciprocal translocations between an autosome and the X chromosome lead to important disruptions in human spermatogenesis. This is explained by the fact that the derivative X chromosome may be interfering in an abnormal sexual vesicle formation, leading to meiotic disturbances and, consequently, to spermatogenetic arrest.
Two males carrying a X-autosome translocation and a severe oligozoospermia have been reported thus far, one carrying a t(X;15)(p11.3;p1) (Fraccaro et al., 1977), the other, a t(X;20)(q10;q10) (Ma et al., 2003). In this latter case, the translocation was inherited from his mother and transmitted to his daughter through ICSI (Ma et al., 2003). Two other males have each fathered a child, one having a t(X;14)(p–;q+) (Buckton et al., 1971), the other, a t(X;1)(p–;q+) (Leichtman et al., 1978).
Mattei et al. (1982) studied the cytogenetic characteristics of the X-autosome translocations. They found that the breakpoints distribution on the X chromosome did not differ significantly from the expected distribution (Mattei et al., 1982). Interestingly, in two subjects each with a balanced t(X;autosome), one of whom had one 46,XY normal son (although paternity was not proven) (Buckton et al., 1971; Leichtman et al., 1978), and in three t(X;autosome) carriers with severe oligozoospermia (Fraccaro et al., 1977; Ma et al., 2003) (our case), the breakpoint on the X chromosome appeared to be always in the pericentromeric region or at the centromere. These balanced translocations may have fewer deleterious effects on spermatogenesis than other X chromosome breakpoints. Thus, when the breakpoint is located near the centromere of the X chromosome, a few germinal cells could be able to complete the meiotic process by producing spermatozoa.
To our knowledge, this is the first cytogenetic study of the chromosomal content in the spermatozoa of a man with a X-autosome translocation. Solely, a small number of spermatozoa could be analysed because of his severe oligozoospermia. The majority of the analysed nuclei showed normal or balanced equipment resulting from alternate segregation with normal (23,Y) or balanced (23,der(X),der(18)) spermatozoa in similar proportions. A 23,Y or a 23,der(X),der(18) spermatozoon fertilizing a normal 23,X oocyte will lead, respectively, to the birth of a 46,XY boy or a 46,X,t(X;18)(q11;p11.1) girl carrying the same translocation as her father and her grandmother. Nevertheless, balanced t(X-autosome) in girls is not necessarily associated with the same phenotype as that of the mother or, as in this case, as that of the grandmother. As proposed by Ma et al. (2003), it is of clinical significance to determine to what degree skewed X chromosome inactivation is present in the resulting female newborns with X-autosome translocation so that the parents can be counselled accordingly (Ma et al., 2003).
The frequency of gametes exhibiting a chromosomal unbalanced equipment was 45.64% with the preferential mode of imbalance being the 3:1 mode. This preferential unbalanced 3:1 segregation is particularly described in the t(11;22)(q23;q11) (Estop et al., 1999; Geneix et al., 2002; Escudero et al., 2003). The same results were found in other reciprocal translocations involving small or acrocentric chromosomes (Geneix et al., 2002; Escudero et al., 2003) or with breakpoints close to the telomeric regions (Martini et al., 1998; Escudero et al., 2003). The 3:1 mode is also favoured when chromosome 9 is involved in the translocation or when, as in our patient, breakpoints are near the centromere (Simpson and Bischoff, 2002; Trappe et al., 2002; Rives et al., 2003; Brugnon et al., 2006).
Theoretically, in the 3:1 mode, gametes with 22 or 24 chromosomes should be found in equal proportions. However, in our case, 15.22% of the spermatozoa had 22 chromosomes and 7.15% had 24 chromosomes. These differences could be explained by a differential viability of the spermatocytes and/or spermatids according to their chromosomal content (Estop et al., 1992; Blanco et al., 1998), but more likely, by the overestimation of the frequency of spermatozoa with 22 chromosomes, possibly due to technical problems (Morel et al., 2004a).
Meiotic segregation patterns of men carrying a balanced reciprocal translocation between two autosomes have been recently reviewed (Morel et al., 2006). The frequency of unbalanced spermatozoa was shown to vary from 19% to >80%. Overall, the frequencies of normal/balanced and unbalanced gametes were, on average, 44% and 56%, respectively. Thus, the risk for X-autosome translocation carriers of producing a chromosomally unbalanced offspring is not higher than that for carriers of a reciprocal translocation between two autosomes, at least in our patient.
During genetic counselling, preimplantation genetic diagnosis (PGD) and conventional prenatal diagnosis were discussed with the couple. As the waiting list for PGD is very long in France, given the frequency of normal or balanced gametes of 
55%, the couple decided to try an in vitro fertilization with sperm microinjection followed by conventional prenatal diagnosis. There were 19 oocytes collected, 12 injected and 6 embryos obtained. One embryo was transferred and a 46,XY newborn baby with no malformation was born.
|
Funding |
|---|
|
TopAbstractIntroductionCase ReportResultsDiscussionFundingReferences |
|---|
A.P. is the recipient of a doctoral studentship from the Région Bretagne.
|
References |
|---|
|
TopAbstractIntroductionCase ReportResultsDiscussionFundingReferences |
|---|
Blanco J, Egozcue J, Clusellas N, Vidal F. 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 (1998) 83:275–280.[CrossRef][Web of Science][Medline]
Brisset S, Izard V, Misrahi M, Aboura A, Madoux S, Ferlicot S, Schoevaert D, Soufir JC, Frydman R, Tachdjian G. Cytogenetic, molecular and testicular tissue studies in an infertile 45,X male carrying an unbalanced (Y;22) translocation: case report. Hum Reprod (2005) 20:2168–2172.
Brugnon F, Van Assche E, Verheyen G, Sion B, Boucher D, Pouly JL, Janny L, Devroey P, Liebaers I, Van Steirteghem A. Study of two markers of apoptosis and meiotic segregation in ejaculated sperm of chromosomal translocation carrier patients. Hum Reprod (2006) 21:685–693.
Buckton KE, Jacobs PA, Rae LA, Newton MS, Sanger R. An inherited X-autosome translocation in man. In: Ann Hum Genet (1971) 35:171–178.[Web of Science][Medline]
Caer E, Perrin A, Douet-Guilbert N, Amice V, De Braekeleer M, Morel F. Different mechanisms of meiotic segregation in psermatozoa from three carriers of a pericentric inversion of chromosome 8. Fertil Steril (2007) June 29 (Epub ahead of print).
Cantu JM, Diaz M, Moller M, Jimenez-Sainz M, Sandoval L, Vaca G, Rivera H. Azoospermia and duplication 3qter as distinct consequences of a familial t(X;3)(q26;q13.2). Am J Med Genet (1985) 20:677–684.[CrossRef][Web of Science][Medline]
De Braekeleer M, Dao TN. Cytogenetic studies in male infertility: a review. Hum Reprod (1991) 6:245–250.
De Braekeleer M, Perrin A, Morel F. Chromosomal abnormalities in male infertility. In: Cytogenetics and Infertility.—De Braekeleer M, ed. (2006) Trivandrum, India: Transworld Research Network. 27–52.
Douet-Guilbert N, Le Bris MJ, Amice V, Marchetti C, Delobel B, Amice J, De Braekeleer M, Morel F. Interchromosomal effect in sperm of males with translocations: report of 6 cases and review of the literature. Int J Androl (2005) 28:372–379.[CrossRef][Web of Science][Medline]
Escudero T, Abdelhadi I, Sandalinas M, Munne S. Predictive value of sperm fluorescence in situ hybridization analysis on the outcome of preimplantation genetic diagnosis for translocations. Fertil Steril (2003) 79:1528–1534.[CrossRef][Web of Science][Medline]
Estop AM, Levinson F, Cieply KM, Van Kirk V. The segregation of a translocation t(1;4) in two male carriers heterozygous for the translocation. Hum Genet (1992) 89:425–429.[Web of Science][Medline]
Estop AM, Cieply KM, Munne S, Feingold E. Multicolor fluorescence in situ hybridization analysis of the spermatozoa of a male heterozygous for a reciprocal translocation t(11;22)(q23;q11). Hum Genet (1999) 104:412–417.[CrossRef][Web of Science][Medline]
Fraccaro M, Maraschio P, Pasquali F, Scappaticci S. Women heterozygous for deficiency of the (p21 leads to pter) region of the X chromosome are fertile. Hum Genet (1977) 39:283–292.[CrossRef][Web of Science][Medline]
Gabriel-Robez O, Rumpler Y, Ratomponirina C, Petit C, Levilliers J, Croquette MF, Couturier J. Deletion of the pseudoautosomal region and lack of sex-chromosome pairing at pachytene in two infertile men carrying an X;Y translocation. Cytogenet Cell Genet (1990) 54:38–42.[Web of Science][Medline]
Geneix A, Schubert B, Force A, Rodet K, Briançon G, Boucher D. Sperm analysis by FISH in a case of t(17;22)(q11;q12) balanced translocation: case report. Hum Reprod (2002) 17:325–331.
Giltay JC, Kastrop PMM, Tiemessen CHJ, Van Inzen WG, Scheres JMJC, Pearson PL. Sperm analysis in a subfertile male with a Y;16 translocation, using four-color FISH. Cytogenet Cell Genet (1999) 84:67–72.[CrossRef][Web of Science][Medline]
Hsu LYF. Phenotype/karyotype correlations of Y chromosome aneuploidy with emphasis on structural aberrations in postnatally diagnosed cases. Am J Med Genet (1994) 53:108–140.[CrossRef][Web of Science][Medline]
Ishikawa T, Konho Y, Yamaguchi K, Oba T, Sakamoto Y, Takenaka A, Fujisawa M. An unusual reciprocal X-autosome translocation in an infertile azoospermic man. In: Fertil Steril (2007) Epub ahead of print.
Kalz-Fuller B, Sleegers E, Schwanitz G, Schubert R. Characterization, phenotypic manifestations and X-inactivation pattern in 14 patients with X-autosome translocations. Clin Genet (1999) 55:362–366.[Web of Science][Medline]
Lee S, Lee SH, Chung TG, Kim HJ, Yoon TK, Kwak IP, Park SH, Cha WT, Cho SW, Cha KY. Molecular and cytogenetic characterization of two azoospermic patients with X-autosome translocation. J Assist Reprod Genet (2003) 20:385–389.[CrossRef][Web of Science][Medline]
Leichtman DA, Schmickel RD, Gelehrter TD, Judd WJ, Woodbury MC, Mellinger KL. Familial Turner syndrome. Ann Intern Med (1978) 89:473–476.
Ma S, Ho Yuen B, Penaherrera M, Koehn D, Ness L, Robinson W. ICSI and the transmission of X-autosomal translocation: a three-generation evaluation of X;20 translocation: case report. Hum Reprod (2003) 18:1377–1382.
Martini E, Von Bergh ARM, Coonen E, de Die-Smulders CEM, Hopman AHN, Ramaekers FCS, Geraedts JPM. Detection of structural abnormalities in spermatozoa of a translocation carrier t(3;11)(q27.3;q24.3) by triple FISH. Hum Genet (1998) 102:157–165.[CrossRef][Web of Science][Medline]
Mattei MG, Mattei JF, Ayme S, Giraud F. X-autosome translocations: cytogenetic characteristics and their consequences. Hum Genet (1982) 61:295–309.[CrossRef][Web of Science][Medline]
Mennicke K, Diercks P, Schlieker H, Bals-Pratsch M, Al Hasani S, Diedrich K, Schwinger E. Molecular cytogenetic diagnostics in sperm. Int J Androl (1997) 20:11–19.[Medline]
Morel F, Fellman F, Roux C, Bresson JL. Meiotic segregation analysis by FISH investigation of spermatozoa of a 46,Y,der(X)t(X;Y) (qter->p22::q11->qter) carrier. Cytogenet Cell Genet (2001) 92:63–68.[CrossRef][Web of Science][Medline]
Morel F, Douet-Guilbert N, Le Bris MJ, Herry A, Amice V, Amice J, De Braekeleer M. Meiotic segregation of translocations during male gametogenesis. Int J Androl (2004) a 27:200–212.[CrossRef][Web of Science][Medline]
Morel F, Douet-Guilbert N, Roux C, Tripogney C, Le Bris MJ, De Braekeleer M, Bresson JL. Meiotic segregation of a t(7;8)(q11.21;cen) translocation in two carrier brothers. Fertil Steril (2004) b 81:682–685.[CrossRef][Web of Science][Medline]
Morel F, Douet-Guilbert N, Perrin A, Le Bris MJ, Amice V, Amice J, De Braekeleer M. Chromosomal abnormalities in spermatozoa. In: Cytogenetics and Infertility.—De Braekeleer M, ed. (2006) Trivandrum, India: Transworld Research Network. 53–112.
Morel F, Laudier B, Guerif F, Couet ML, Royere D, Roux C, Bresson JL, Amice V, De Braekeleer M, Douet-Guilbert N. Meiotic segregation analysis in spermatozoa of pericentric inversion carriers using fluorescence in-situ hybridization. Hum Reprod (2007) 22:136–141.
Panasiuk B, Usinskiene R, Kostyk E, Rybalko A, Stasiewicz-Jarocka B, Krzykwa B, Pienkowska-Grela B, Kucinskas V, Michalova K, Midro AT. Genetic counselling in carriers of reciprocal chromosomal translocations involving the short arm of chromosome X. Ann Génét (2004) 47:11–28.[Web of Science][Medline]
Pinho MJ, Neves R, Costa P, Ferras C, Sousa M, Alves C, Almeida C, Fernandes S, Silva J, Ferras L, et al. Unique t(Y;1)(q12;q12) reciprocal translocation with loss of the heterochromatic region of chromosome 1 in a male with azoospermia due to meiotic arrest: a case report. Hum Reprod (2005) 20:689–696.
Quack B, Speed RM, Luciani JM, Noel B, Guichaoua M, Chandley AC. Meiotic analysis of two human reciprocal X-autosome translocations. Cytogenet Cell Genet (1988) 48:43–47.[Web of Science][Medline]
Rives N, Jarnot M, Mousset-Siméon N, Joly G, Mace B. Fluorescence in situ hybridisation (FISH) analysis of chromosome segregation and interchromosomal effect in spermatozoa of a reciprocal translocation t(9;10)(q11;p11.1) carrier. J Hum Genet (2003) 48:535–540.[CrossRef][Web of Science][Medline]
Simpson JL, Bischoff F. Genetic counseling in translocations. Urol Clin N Am (2002) 29:793–807.[CrossRef][Web of Science][Medline]
Solari AJ, Rahn IM, Ferreyra ME, Carballo MA. The behavior of sex chromosomes in two human X-autosome translocations: failure of extensive X-inactivation spreading. Biocell (2001) 25:155–166.[Web of Science][Medline]
Taiar N, Qumsiyeh MB, Croteau S, Rollet J, Benkhalifa M. Detection of t(X;Y) in 2 XX males using fluorescent in situ hybridization. Ann Génét (1995) 38:102–105.[Web of Science][Medline]
Trappe R, Bohm D, Kohlhase J, Weise A, Liehr T, Essers G, Meins M, Zoll B, Bartels I, Burfeind P. A novel family-specific translocation t(2;20)(p24.1;q13.1) associated with recurrent abortions: molecular characterization and segregation analysis in male meiosis. Cytogenet Genome Res (2002) 98:1–8.[CrossRef][Web of Science][Medline]
Yamada K, Nanko S, Hattori S, Isurugi K. Cytogenetic studies in a Y-to-X translocation observed in three members of one family, with evidence of infertiliy in male carriers. Hum Genet (1982) 60:85–90.[CrossRef][Web of Science][Medline]
Submitted on April 21, 2007; resubmitted on July 30, 2007; accepted on September 5, 2007.
CiteULike 
Connotea 
Del.icio.us What's this?
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||