1 Service de Génétique et Reproduction and 2 Service de Gynécologie-Obstétrique, Hôpital Antoine Béclère, 92140 Clamart, 3 Service d'Urologie-Andrologie, 4 Laboratoire d'Hormonologie et de Biologie Moléculaire, 5 Service d'Anatomie et Cytologie Pathologiques and 6 Laboratoire de Biologie Andrologique, Hôpital Bicêtre, 94270 Le Kremlin Bicêtre, France
7 To whom correspondence should be addressed at: Service de Biologie et Génétique de la Reproduction, Hôpital Antoine Béclère, 157, rue de la Porte de Trivaux, 92140 Clamart, France. Email: sophie.brisset{at}abc.ap-hop-paris.fr
![]() |
Abstract |
---|
![]() ![]() ![]() ![]() ![]() ![]() |
---|
Key words: AZF deletions/azoospermia/genotypephenotype correlation/testicular biopsy/(Y;autosome) translocation
![]() |
Introduction |
---|
![]() ![]() ![]() ![]() ![]() ![]() |
---|
![]() |
Case report |
---|
![]() ![]() ![]() ![]() ![]() ![]() |
---|
Conventional cytogenetic analysis
Chromosome analyses were performed on cultured lymphocytes. Standard techniques were used for the preparation of metaphase spreads. The constitutional karyotype was established after R banding.
Fluorescence in situ hybridization (FISH)
FISH analyses were performed on lymphocyte metaphase spreads from the patient using standard protocols. The following DNA probes were used for FISH analysis: X chromosome (DXZ1), Y chromosome (DYZ3) and chromosome 14/22 (D14Z1/D22Z1) centromeric specific probes, whole chromosome painting probes for chromosome 22 and Y chromosome and DiGeorge N25 chromosome region probe (Vysis, USA).
Molecular analysis
DNA was extracted from blood lymphocytes. Microdeletion analysis of the Y chromosome was performed using a sequence-tagged site (STS)PCR approach. Twenty-eight STS corresponding to the three distinct AZF loci were selected. The STS tested were sY84 for AZFa, sY95, sY97, sY169, sY102, sY105, sY109 for the interval between AZFa and AZFb, sY113, sY115, sY117, sY124, sY130, sY134, sY136, sY143, sY142 for AZFb, sY152, sY232, sY156, sY240, sY148, sY249, sY204, sY208, sY254, sY269, sY158 for AZFc, and sY160 for the heterochromatic distal Yq region.
Testicular tissue analyses
Fresh testicular tissue was obtained surgically from the patient. Direct cytological examination and histological examination were performed on testicular tissue.
Immediately after testicular biopsy, the extracted testicular seminiferous tubules were deposited on a sterile glass slide and gently dissected with sterile microscissors in FertiCult medium (Izard et al., 1999). The wet preparation of the suspension was then checked for the presence of sperm under a phase-contrast microscope at x400 magnification. The morphology of sperm was assessed using Shorr staining. Sperm morphology was evaluated according to modified David's classification (Auger et al., 2001
).
For testicular histology, the biopsy specimen was fixed with AFA (75% alcohol, 0.8% formalin and 5% acetic acid). After embedding with paraffin, tissue sections were cut with a thickness of 4 µm. All sections were stained with haematoxylin, eosin and saffron.
Conventional cytogenetic analysis
The cytogenetic analysis was initially interpreted as a 45,X karyotype. Further cytogenetic examination revealed a mosaic 45,X[15]/46,X, + mar[3] with a small chromosomal marker whose origin could not be resolved by conventional analysis (Figure 1A). The father had a normal karyotype.
|
The 14/22 -satellite centromeric probe showed signals on both centromeres of chromosomes 14, one signal on the centromere of one chromosome 22 and one signal on the centromere of the small chromosome marker. This suggested that the small marker chromosome included the centromere of one chromosome 22 (Figure 1C).
Two-colour FISH with paint specific probes for Y chromosome (green) and chromosome 22 (red) showed a derivative Y chromosome containing the short arm, the centromere and a small proximal part of the long-arm euchromatin of the Y chromosome and the long arm of a chromosome 22. Thus, the small marker chromosome represented a part of the translocation event corresponding to the short arm and the centromere of chromosome 22 (Figure 1D). On chromosome 22, the breakpoint occurred in the region just below the centromere as confirmed by FISH using the DiGeorge locus probe. This probe hybridized on both chromosome 22 and derivative Y chromosome, indicating therefore that the breakpoint on chromosome 22 was located between centromere and 22q11.2 region (data not shown).
The results of these hybridization experiments indicated therefore that the patient was a carrier of an unbalanced translocation t(Y;22)(q11.2;q11.1) (Figure 2).
|
Testicular tissue analyses
A wet preparation of the testicular tissue sample showed the presence of non-motile sperm. Shorr staining showed a normal morphology (the normal morphology rate was 34%) (Figure 3).
|
![]() |
Discussion |
---|
![]() ![]() ![]() ![]() ![]() ![]() |
---|
(Y;autosome) translocations have been associated with normal and abnormal spermatogenesis. Hsu (1994) reviewed over 130 cases of (Y;autosome) translocations. In the most common form, the heterochromatic region of Yq (Yqh or Yq12) is translocated onto the short arm of an acrocentric chromosome (D or G group chromosome). Other types of (Y;autosome) translocation include balanced and unbalanced reciprocal translocations. A chromosomal breakpoint at band Yq11, as observed in our case, is rarely described. Twenty cases of adult males carrying a (Y;autosome) translocation with a breakpoint at Yq11 region are reviewed in Table I. Since the phenotype may depend on the localization of the Yq breakpoint and on the nature of Yq material lost, it is of great interest to ascertain the Yq breakpoint by DNA molecular studies (AZF screening or interval deletions studies). Deletions of the AZFa, b and c regions are associated with abnormal spermatogenesis from SCOS to hypospermatogenesis (Vogt et al., 1996
; Krausz et al., 2003
; Simoni et al., 2004
).
|
The sperm could thereafter be used for ICSI in combination with preimplantation genetic diagnosis. The chromosomal risks of this translocation in offspring are aneuploidy of chromosome 22 and sex numerical anomalies. Nevertheless, the most important genetic risk for the couple concerns sex chromosome anomalies because monosomy or trisomy 22 are not viable.
In conclusion, our case highlights the importance of documenting (Y;autosome) translocations with molecular and testicular tissue analyses in order to establish a genotypephenotype correlation.
![]() |
References |
---|
![]() ![]() ![]() ![]() ![]() ![]() |
---|
Andersson M, Page DC, Pettay D, Subrt I, Turleau C, de Grouchy J and de la Chapelle A (1988) Y;autosome translocations and mosaicism in the aetiology of 45,X maleness: assignment of fertility factor to distal Yq11. Hum Genet 79, 27.[CrossRef][ISI][Medline]
Arnemann J, Schnittger S, Hinkel GK, Tolkendorf E, Schmidtke J and Hansmann I (1991) A sterile male with 45,X0 and a Y;22 translocation. Hum Genet 87, 134138.[CrossRef][ISI][Medline]
Auger J, Eustache F, Andersen AG, Irvine DS, Jorgensen N, Skakkebaek NE, Suominen J, Toppari J, Vierula M and Jouannet P (2001) Sperm morphological defects related to environment, lifestyle and medical history of 1001 male partners of pregnant women from four European cities. Hum Reprod 16, 27102717.
Bardoni B, Zuffardi O, Guioli S, Ballabio A, Simi P, Cavalli P, Grimoldi MG, Fraccaro M and Camerino G (1991) A deletion map of the human Yq11 region: implications for the evolution of the Y chromosome and tentative mapping of a locus involved in spermatogenesis. Genomics 11, 443451.[ISI][Medline]
Brandell RA, Mielnik A, Liotta D, Ye Z, Veeck LL, Palermo GD and Schlegel PN (1998) AZFb deletions predict the absence of spermatozoa with testicular sperm extraction: preliminary report of a prognostic genetic test. Hum Reprod 13, 28122815.
Buonadonna AL, Cariola F, Caroppo E, Di Carlo A, Fiorente P, Valenzano MC, D'Amato G and Gentile M (2002) Molecular and cytogenetic characterization of an azoospermic male with a de-novo Y;14 translocation and alternate centromere inactivation. Hum Reprod 17, 564569.
Callen DF, Sutherland GR and Carter RF (1987) A fertile man with tdic(Y;22): how a stable neo-X1X2Y sex-determining mechanism could evolve in man. Am J Med Genet Suppl 3, 151155.[Medline]
De Braekeleer M and Dao TN (1991) Cytogenetic studies in male infertility: a review. Hum Reprod 6, 245250.[ISI][Medline]
Delobel B, Djlelati R, Gabriel-Robez O, Croquette MF, Rousseaux-Prevost R, Rousseaux J, Rigot JM and Rumpler Y (1998) Y-Autosome translocation and infertility: usefulness of molecular, cytogenetic and meiotic studies. Hum Genet 102, 98102.[CrossRef][ISI][Medline]
Dutrillaux B and Gueguen J (1975) Mitotic and meiotic analysis of an Y-autosome translocation. Humangenetik 27, 241245.[CrossRef][ISI][Medline]
Faed MJ, Lamont MA and Baxby K (1982) Cytogenetic and histological studies of testicular biopsies from subfertile men with chromosome anomaly. J Med Genet 19, 4956.[Abstract]
Gal A, Weber B, Neri G, Serra A, Muller U, Schempp W and Page DC (1987) A 45,X male with Y-specific DNA translocated onto chromosome 15. Am J Hum Genet 40, 477488.[ISI][Medline]
Giltay JC, Tiemessen CH, van Inzen WG and Scheres JM (1998) One normal child and a chromosomally balanced/normal twin after intracytoplasmic sperm injection in a male with a de-novo t(Y;16) translocation. Hum Reprod 13, 27452747.
Giltay JC, Kastrop PM, Tiemessen CH, van Inzen WG, Scheres JM and Pearson PL (1999) Sperm analysis in a subfertile male with a Y;16 translocation, using four-color FISH. Cytogenet Cell Genet 84, 6772.[CrossRef][ISI][Medline]
Gonzales J, Lesourd S and Dutrillaux B (1981) Mitotic and meiotic analysis of a reciprocal translocation t(Y;3) in an azoospermic male. Hum Genet 57, 111114.[ISI][Medline]
Hopps CV, Mielnik A, Goldstein M, Palermo GD, Rosenwaks Z and Schlegel PN (2003) Detection of sperm in men with Y chromosome microdeletions of the AZFa, AZFb and AZFc regions. Hum Reprod 18, 16601665.
Hsu LY (1994) Phenotype/karyotype correlations of Y chromosome aneuploidy with emphasis on structural aberrations in postnatally diagnosed cases. Am J Med Genet 53, 108140.[ISI][Medline]
Izard V, Marmor D, Peron A, Paradis V, Benoit G, Soufir JC and Jardin A (1999) Cytological evaluation of germinal populations as part of a testicular sperm retrieval strategy. In Hamamah S, Mieusset R, Olivennes F, and Frydman R (eds) Male Sterility and Motility Disorders. Etiological Factors and Treatment. Serono Symposia, Norwell, USA, pp. 253258.
Jarow JP, Espeland MA and Lipshultz LI (1989) Evaluation of the azoospermic patient. J Urol 142, 6265.[ISI][Medline]
Krausz C, Quintana-Murci L and McElreavey K (2000) Prognostic value of Y deletion analysis: what is the clinical prognostic value of Y chromosome microdeletion analysis? Hum Reprod 15, 14311434.
Krausz C, Forti G and McElreavey K (2003) The Y chromosome and male fertility and infertility. Int J Androl 26, 7075.[CrossRef][ISI][Medline]
Laurent C and Dutrillaux B (1976) Translocation t(Y;14) in an azoospermic man. Ann Genet 19, 207209.[ISI][Medline]
Laurent C, Chandley AC, Dutrillaux B and Speed RM (1982) The use of surface spreading in the pachytene analysis of a human t (Y;17) reciprocal translocation. Cytogenet Cell Genet 33, 312318.[ISI][Medline]
Pabst B, Glaubitz R, Schalk T, Schneider U, Schulze W and Miller K (2002) Reciprocal translocation between Y chromosome long arm euchromatin and the short arm of chromosome 1. Ann Genet 45, 58.[ISI][Medline]
Ratomponirina C, Couturier J, Gabriel-Robez O, Rumpler Y, Dutrillaux B, Croquette M, Rabache Q and Leduc M (1985) Aberrations of the synaptonemal complexes in a male 46,XY,-14, + der(14)t(Y;14). Ann Genet 28, 214218.[ISI][Medline]
Sasagawa I, Nakada T, Adachi Y, Kato T, Sawamura T, Ishigooka M and Hashimoto T (1993) Y-autosome translocation associated with azoospermia. Scand J Urol Nephrol 27, 285288.[ISI][Medline]
Schempp W, Weber B, Serra A, Neri G, Gal A and Wolf U (1985) A 45,X male with evidence of a translocation of Y euchromatin onto chromosome 15. Hum Genet 71, 150154.[ISI][Medline]
Silber SJ, Alagappan R, Brown LG and Page DC (1998) Y chromosome deletions in azoospermic and severely oligozoospermic men undergoing intracytoplasmic sperm injection after testicular sperm extraction. Hum Reprod 13, 33323337.[Abstract]
Simoni M, Bakker E and Krausz C (2004) EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions. State of the art 2004. Int J Androl 27, 240249.[CrossRef][ISI][Medline]
Smith A, Fraser IS and Elliott G (1979) An infertile male with balanced Y;19 translocation. Review of Y;autosome translocations. Ann Genet 22, 189194.[ISI][Medline]
Subrt I and Blehova B (1974) Robertsonian translocation between the chromosome Y and 15. Humangenetik 23, 305309.[CrossRef][ISI][Medline]
Teyssier M, Rafat A and Pugeat M (1993) Case of (Y;1) familial translocation. Am J Med Genet 46, 339340.[CrossRef][ISI][Medline]
Turleau C, Chavin-Colin F and de Grouchy J (1980) A 45,X male with translocation of euchromatic Y chromosome material. Hum Genet 53, 299302.[ISI][Medline]
Van Assche E, Bonduelle M, Tournaye H, Joris H, Verheyen G, Devroey P, Van Steirteghem A and Liebaers I (1996) Cytogenetics of infertile men. Hum Reprod 11 (Suppl 1), 124.
Vogt PH (2004) Genomic heterogeneity and instability of the AZF locus on the human Y chromosome. Mol Cell Endocrinol 224, 19.[CrossRef][ISI][Medline]
Vogt PH, Edelmann A, Kirsch S, Henegariu O, Hirschmann P, Kiesewetter F, Kohn FM, Schill WB, Farah S, Ramos C et al (1996) Human Y chromosome azoospermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet 5, 933943.
Submitted on September 16, 2004; resubmitted on March 22, 2005; accepted on March 31, 2005.
|