BJMG 11/1 (2008) 51-54 10.2478/v10034-008-0017-x

ORIGINAL ARTICLE

A 45,X/47,XYY/46,XY KARYOTYPE AND Y MICRODELETION IN AN INFERTILE MALE

Bulakbasi T1, Sahin FI1,*, Yil maz Z1, Zeyneloglu HB2

Corresponding Author: Professor Dr. Feride Iffet Sahin, Department of Medical , Faculty of , Baskent University, Kubilay Sokak No:36, Maltepe 06570, Ankara, Turkey; Tel : +90-312-2324400; Fax : +90-312-2319134; E-mail: [email protected]

ABSTRACT INTRODUCTION

Infertility affects 10-15% of all couples and the Infertility is an important health problem that male factor is responsible in approximately half of affects about 10-15% of all couples attempting them. The advances in assisted reproduction tech- pregnancy [1,2]. abnormalities niques and genetic diagnoses have increased the are an important cause of male infertility [1,2]. In- percentage of couples who are able to conceive. We tracytoplasmic sperm injection (ICSI) is an effective report on an infertile male patient with , therapeutic method, especially in couples with male referred to our laboratory before intra­cytoplasmic infertility and/or unsuccessful in vitro fertilization sperm injection (ICSI), whose peripheral lympho- (IVF) experiences [1]. Cytogenetic evaluation of cyte karyotype showed a 45,X/46,XY mosaicism. couples prior to ICSI shows that both genders are A (Yq) microdeletion was detected equal­ly at risk for sex chromosome abnormalities. in the azoospermia factor (AZF) regions, AZFb and An increased incidence of the 47,XYY karyotype in AZFc. The karyotype from testicular tissue cultures infertile males has been reported [2], although the was 45,X[13]/47,XYY[12]/46,XY[25]. Flu- majority of individuals with this karyotype are fer- orescence in situ hybridization (FISH) performed tile and show normal spermatogenesis­ [2,3]. In those on blood lymphocytes and testicular cells showed a males who have spermatogenetic failure, abnormal- 47,XYY line in both, in 8 and 23% of cells, re- ities of azoospermia factor in the euchromatic spectively. Histological examination of the testicu- region on the long arm of the Y chromosome (Yq) lar biopsy revealed very few seminiferous tubules, have been reported [2]. The Yq microdeletions of which were hyalinized and composed only of Ser- azoospermia factor (AZF) regions are major causes toli cells. Intracytoplasmic sperm injection was not of infertility associated with severe oligospermia suggested to the patient due to the sex chromosome and azoo­spermia [4] and may also be associated mosaicism, Yq micro­deletion and biopsy findings. with and germinal gonosomal mosaicism [5-7]. Key words: Male infertility; Sex chromosome mosa­icism; Y Microdeletion (Yq) CASE REPORT

A 42-year-old infertile male patient with non ob- 1 Department of , Faculty of Medi- structive azoospermia was referred to our genetics cine, Baskent University, Ankara, Turkey laboratory for diagnostic studies before ICSI treat- 2 Department of Genycology and , Faculty ment. In prior IVF attempts no mature spermatozoa of Medicine, Baskent University, Ankara, Turkey

51 45,X/47,XYY/46,XY AND Y DELETION were found, so the procedure was carried out with have a broad spectrum of phenotypic effects [2]. spermatid microinjection and failed. The patient Our patient revealed a 45,X/46,XY mosaicism had no significant family history and had undergone during routine blood analysis. This karyotype was varicocelectomy 5 years previously. In sperm analy- found in seven cases in the same study [2]. Our pa- sis, the seminal volume was 2.7 mL and no motile or tient was infertile because of azoospermia, but he immotile sperm was seen. Chromosome analysis of also had a Yq microdeletion covering the AZFb and cultured blood lymphocytes showed a mosaic kary- AZFc regions. It has been reported that numerical Y otype 45,X[32]/46,XY[68] and the karyotype inter- chromosomal defects may accompany Yq microde- pretation was made according to ISCN 2005 [8]. A letions, as deleted regions tend to be lost during Yq microdeletion analysis was performed, screening as a result of mitotic instability [6,7]. for two deletions in AZFa region, five in AZFb, four There is a close association between large Yq de- in AZFc and one in AZFd, and for ZFY and SRY letions and gonosomal mosaicism in both somatic genes. The sY277 and sY283 regions of the DAZ and germinal cells [5,7]. As in our case, mosaicism in AZFc, sY131 and sY143 regions in AZFb, in germinal cells may remain undetected unless a were found to be deleted. Histological­ examination specific analysis is performed. We first detected the of the testicular biopsy revealed hyalinization­ in 47,XYY cell line cytogenetically in testicular tissue the few seminiferous tubules present and they were and confirmed its presence by FISH analysis in 23% composed only of Sertoli cells. No germinal epi- of these cells. The FISH analysis of peripheral lym- thelium or spermatogenesis was detected. The cells phocytes revealed the same finding in 8% of cells, from the testicular biopsy culture showed the karyo- which were not identified by conventional cytoge- type 45,X[13]/ 47,XYY[12]/46,XY[25]. To further netics. investigate the new cell line in blood lymphocytes The cases of sex chromosome mosaicism re- and to evaluate larger number of cells, ported in the literature show great diversity, from in situ hybridization (FISH) was performed on both to patients with ambiguous geni- tissues using the centromeric probes, Y chromosome talia [9]. The karyotypes with 45,X and 47,XYY DYZ3 and DXZ1 (chromosome cell lines could both present as males and X/Y cocktail probe; Qbiogene Inc., Carlsbad, CA, phenotypically and features like short stature or go- USA). Two hundred cells were analyzed from each nadal tumors have also been reported [9]. Our pa- tissue and the XYY signal was detected in 8% of tient is an apparently normal male, except for the the lymphocytes and in 23% of the testicular cells. azoo­spermia which could be due to the coexistence The patient was informed about his condition dur- of a Yq microdeletion. The dominant cell line pres- ing genetic counseling. Due to the sex chromosome ent in different tissues correlate with the cases re- mosaicism, Yq microdeletion and biopsy findings, ported before. Our patient had different percentages ICSI was not suggested. Sperm extraction from tes- of the 47,XYY cell line in different tissues, as ex- tis was performed, but the procedure produced no pected in these mosaic states, and this variation may spermatids. explain the diversity of the seen in these patients [3,9]. DISCUSSION The formation of 45,X/47,XYY/46,XY mosa- icism may be caused by at least two mechanisms. The relationship between infertility and chro- The first is paternal non disjunction at II mosomal abnormalities has been well documented followed by loss of the chromosome in subsequent over the past 25 years [2]. Since an increase in chro- mitoses. The second is a post-zygotic mitotic error mosomal abnormalities correlates with a decrease which explains the different amounts of mosaicism in sperm count, abnormalities in sperm count are present in different tissues [3]. Either mechanism the most important indications for chromosome could be the cause of mosaicism in our patient, analysis in infertile males [2]. Numerical sex chro- while the presence of Yq microdeletion makes him mosome aberrations constitute a small percentage especially interesting. of the anomalies; since only 3.32% were reported When sex chromosomal is detected in a study of 2,196 infertile men; and they tend to in infertile males, it should be remembered that Yq

52 BALKAN JOURNAL OF MEDICAL GENETICS Bulakbasi T1, Sahin FI1,*, Yilmaz Z1, Zeyneloglu HB2 micro­deletions may accompany the clinical picture point to disruption of AZF, a spermatogen- as they may induce mitotic instability of the Y chro- esis gene. Hum Genet 1992; 89(5): 491-496. mosome, and cause mosaicism that may be unde- 5. Siffroi JP, Le Bourhis C, Krausz C, Barbaux tected in blood cells, but make considerable contri- S, Quintana-Murci L, Kanafani S Rouba H, Bujan bution to the germinal tissue. Evaluation of infertile L, Bour­rouillou G, Seifer I, Boucher D, Fellous M, males with spermatogenetic defects for gonosomal McElreavey K, Dadoune JP. Sex chromosome mo- mosaicism could lead to a better assessment for es- saicism in males carrying Y chromosome long arm timating the outcome of assisted reproduction tech­ deletions. Hum Reprod 2000; 15(12): 2559-2562. niques. This case demonstrates the use of testicular 6. Jaruzelska J, Korcz A, Wojda A, Jedrzejcak biopsy material as another tissue to be karyotyped. P, Bierla J, Surmacz T, Pawelczyk L, Page DC, Ko- The second tissue in this case revealed a third line tecki M. Mosaicism for 45,X cell line may accen- of cells with gono­somal aberration and contributed tuate the severity of spermatogenic defects in men to predicting an outcome and shaping the genetic with AZFc deletion. J Med Genet 2001; 38(11): counseling for this patient. 798-802. 7. Patsalis PC, Skordis N, Sismani C, Kousouli- REFERENCES dou L, Koumbaris G, Eftychi C, Stavrides G, Ioulia- nos A, Kitsiou-Tzeli S, Galla-Voumvouraki A, Ko- 1. Morel F, Gallon F, Amice V, Le Bris MJ, Le smaidou Z, Hadjiathanasiou CG, McElreavey K. Martelot MT, Roche S, Valéri A, Derrien V, Herry Identification of high frequency of Y chromosome A, Amice J, De Braekeleer M. Sex chromosome deletions in patients with sex chromosome mosa- mosaicism in couples undergoing intracytoplasmic icism and correlation with the clinical sperm injection. Hum Reprod 2002; 17(10): 2552- and Y-. Am J Med Genet A 2555. 2005; 135(2): 145-149. 2. Gekas J, Thepot F, Turleau C, Siffroi JP, 8. ISCN 2005: an international system for hu- Dadoume JP, Wasels R Benzacken B. Association cyto­genetic nomenclature (2005): recommen- des Cyto­geneticiens de Langue Francaise. Chromo- dations of the International Standing Committee of somal factors of infertility in candidate couples for Human Cytogenetic Nomenclature. In: Shaffer LG, ICSI: an equal risk of constitutional aberrations in Tommerup N, Eds. Basel: S. Karger, 2005. women and men. Hum Reprod 2001; 16(1): 82-90. 9. Monastirli A, Stephanou G, Georgiou S, 3. Robinson DO, Jacons PA. The origin of the Andriano­poulos C, Pasmatzi E, Chroni E Katrivanou extra Y chromosome in males with a 47,XYY kary- A, Dimo­poulos P, Demopoulos NA, Tsambaos D. otype. Hum Mol Genet 1999; 8(12): 2205-2209. Short stature, type E brachydactyly, exostoses, gy- 4. Vogt PH, Chandley AC, Hargreave TB, Keil necomastia, and crypt­orchidism in a patient with R, Ma K, Sharkey A. Microdeletions in interval 6 of 47,XYY/45,X/46,XY mosa­icism. Am J Med Sci the Y chromosome of males with idiopathic sterility 2005; 329(4): 208-210.

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