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Infertile spermatozoa in a human carrier of robertsonian translocation 14;22
FERTILITY AND STERILITY威 VOL. 78, NO. 5, NOVEMBER 2002 Copyright ©2002 American Society for Reproductive Medicine Published by Elsevier Science Inc. Printed on acid-free paper in U.S.A.
Infertile spermatozoa in a human carrier of robertsonian translocation 14;22 Baccio Baccetti, M.D.,a,b,c Serena Capitani, M.D., a,b,c Giulia Collodel, M.D.,a Mariela Estenoz, B.S.,b Laura Gambera, Ph.D.,a and Paola Piomboni, M.D.a,c University of Siena and Azienda Ospedaliera Senese (A.O.S.) Hospital, Siena, Italy
Received December 13, 2001; revised and accepted March 8, 2002. Supported by the Ministry of the University, Scientific and Technological Research and the University of Siena, MURST-COFIN 2000, and A.O.S. Hospital, Siena, Italy. Reprint requests: Baccio Baccetti, Department of Pediatrics, Obstetrics, and Reproductive Medicine, Section of Biology, via T. Pendola 62, 53100 Siena, Italy (FAX: 39-0577-233509; E-mail: [email protected]). a Department of Pediatrics, Obstetrics, and Reproductive Medicine, Section of Biology, University of Siena. b National Research Council, Center for the Study of Germinal Cells, A.O.S. Hospital. c Regional Reference Center for Male Infertility, A.O.S. Hospital. 0015-0282/02/$22.00 PII S0015-0282(02)03379-4
Objective: To present the ultrastructural, functional, and chromosomal analyses of spermatozoa from an infertile man with normal phenotype and chromosomal translocation 14;22. Design: Case report. Setting: Regional Reference Center for Male Infertility in Siena, Italy. Patient(s): A 36-year-old man with primary infertility for 3 years and his parents. Intervention(s): Family history and lymphocytic karyotypes, physical and hormonal assays, and semen analysis. Main Outcome Measure(s): Morphological sperm evaluation was performed by light, fluorescent, and electron microscopy; chromosomal constitution was examined by the fluorescence in situ hybridization (FISH) technique. The penetration ability of spermatozoa was checked by the hamster test. Result(s): The spermatozoa of the patient showed unusual ultrastructural defects. The nuclei were large, spheroidal, and generally uncondensed; the acrosomes were frequently absent or reduced; and the axonemes were often devoid of dynein arms or central singlet tubules. These characteristics are related to immaturity. The lymphocytic karyotype revealed a robertsonian translocation 14;22 in the sterile patient and his mother. FISH sperm analysis demonstrated a high frequency of diploidy for the chromosome 18,XY. The hamster penetration test gave negative results. Conclusion(s): The unusual structural sperm immaturity is associated with the translocation 14;22. This chromosomal anomaly may therefore negatively influence the spermatogenesis; an interchromosomal effect on meiosis segregation is also suggested. (Fertil Steril威 2002;78:1127–30. ©2002 by American Society for Reproductive Medicine.) Key Words: Robertsonian translocation, sperm function, FISH, human infertility, electron microscopy
A number of investigators have suggested that genetic and chromosomal factors can contribute to human male factor infertility, particularly in azoospermic and oligozoospermic men (1). This impairment of spermatogenesis in individuals with chromosomal anomalies has been demonstrated by Chandley et al. (2) and seems to be particularly due to translocations (3, 4). This view was criticized by Marmor et al. (5), who did not find any differences in sperm number, motility, and morphology in the carriers of balanced translocations when compared with individuals with normal karyotypes. In addition, Wyrobek et al. (6) previously substantiated this opinion in studies on mice. These differences of opinion are still maintained, even when the use of electron micros-
copy allows a more accurate analysis of sperm characteristics. In’t Veld et al. (7) and Ogawa et al. (8) correlate extreme oligoasthenozoospermia with the robertsonian translocations 13;14 and 13;13. Matsuda et al. (9) and Testart et al. (10), however, conclude that structural chromosomal anomalies, namely, different balanced autosomal translocations or inversions in the chromosome 2 and 14, seem to be unrelated to sperm motility and morphology. These conflicting opinions may derive from the different methods used by the investigators in assessing the sperm quality. Guichaoua et al. (11) performed a pachytene analysis in a sterile male 14;22 robertsonian translocation carrier. They reported that the patient was oligoasthenozoospermic, with normal sperm morphology at the light microscopy level. 1127
We performed semen analyses by light and electron microscopy and by the fluorescence in situ hybridization (FISH) technique in a sterile male carrier of a robertsonian translocation 14;22, and we found unusual structural and genomic sperm anomalies.
CASE REPORT A 36-year-old man presented to our laboratory for semen analysis after 3 years of sexual intercourse without conception. His wife, also 36, did not have any fertility problems. The patient was an only child as was his mother. Sexual development, medical history, physical examination, and hematic hormonal levels were normal. The patient did not have a ciliary defect. Total T, cortisol, T3, T4, TSH, E2, FSH, LH, prolactin, and inhibin levels were obtained. In 1999, this man underwent assisted fertilization twice. The first time, four oocytes were inseminated by IVF, but no activation was achieved. Seven months later, four more oocytes underwent IVF without sperm penetration. A subsequent intracytoplasmic sperm injection also had negative results. The lymphocytic karyotype of the patient (Fig. 1a) revealed the presence of a robertsonian translocation 45,XY t(14;22). The karyotype analysis of his mother showed the same translocation. Semen analysis was performed three times at 1-month intervals in 1999 and once in 2001. The samples were collected through masturbation after 4 days of sexual abstinence. The ejaculate was fully liquefied and then volume, pH, number, and motility of spermatozoa were evaluated following the World Health Organization (WHO) guidelines (12). The sperm number in the four different examinations was between 43,500,000 and 120,000,000; pH, 7.8; volume, 2.5–3 mL; and sperm progressive motility between 8% and 17%. Severe asthenospermia was found in the four samples. The eosin Y test revealed 28% live spermatozoa. To evaluate the sperm morphology by light microscopy, we used a modified Papanicolau staining, according to the protocol reported in the WHO guidelines (12). In all the samples, we observed that 30% of sperm had normal morphology and that the percentage of spermatozoa, bi- or multinucleated, was very high (27%). We further applied a trichromatic staining (13) with different molecular probes for the simultaneous detection of acrosomal content, chromatin organization, and axonemal texture. The spermatozoa, examined by fluorescent microscopy, showed a nuclear positivity to the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling (TUNEL), revealing DNA fragmentation in 21.3% of the cells; the acrosome was badly shaped or absent in 56% of the cells; the axonemal shape and presence of tubulin resulted in an alteration in 55.25% of the sperm. 1128 Baccetti et al.
Translocation 14;22 in an infertile man
For the evaluation of sperm ultrastructure, carried out on two different samples in 1999 and in 2001, we performed transmission electron microscopic (TEM) analysis on ejaculated spermatozoa as reported in Baccetti et al. (14). These observations of 300 suitable sperm sections, cut from several areas of the ejaculate, point out peculiar anomalies of the head structure, characteristic of sperm immaturity (Fig. 1b): the nuclei were irregularly shaped, often elliptical or spherical, with uncondensed or marginated chromatin (structural characteristic of apoptosis). Moreover, spermatozoa binucleate or multinucleate were frequently detected. The acrosome was absent in half of the analyzed spermatozoa; in the other half, this organelle was frequently short and badly shaped. The cytoplasmic residues were present in 50% of the spermatozoa, embedding a disorganized rolled-up axoneme, sometimes coiled around the nucleus. The mitochondrial shape and assembly frequently appeared irregular. Also, in the tails with regular shape (Figs. 1c and d), the axonemal structure was often altered, with absent central tubules (Fig. 1e) and with some doublets having incomplete or absent dynein arms. The accessory fibers and the fibrous sheath were rather well organized in about 50% of the examined tail sections. After the statistical elaboration of TEM data with the formula of Baccetti et al. (14), we found that in the examined samples the mean of the percentages of spermatozoa probably devoid of defects was ⬃0.050%, corresponding to a mean of the total numbers of “healthy spermatozoa” of ⬃50,000. These values are very low. To thoroughly investigate the penetration capacity of the spermatozoa, we performed the hamster test, according to Benet et al. (15). Zona-free hamster oocytes and sperm samples of the patient and two different donors were coincubated. The experiment was repeated twice. Penetration was observed only in the donor’s samples and never in the patient’s samples. Subsequently, we carried out FISH analysis, according to Kovanci et al. (16), to evaluate the aneuploidy frequency in ejaculated spermatozoa. A mix of probes (CEP, Vysis, IL) for chromosomes 18,XY was used. The disomy frequency of the analyzed chromosomes, evaluated in 5,474 spermatozoa, was in the standard range compared with the values found in fertile individuals. The observed frequency of diploidies for the investigated chromosomal sets was very high (5.78% of spermatozoa), despite the fact that the nuclei of these sperm after 6-diamino-2-phenylindole staining were shown to be single, with a well-defined edge.
DISCUSSION In this study, we report the case of a sterile male patient carrier of a robertsonian translocation 14;22 inherited from his mother. The spermatozoa of this patient were examined by several techniques, including light, fluorescent, and elecVol. 78, No. 5, November 2002
FIGURE 1 a, The lymphocytic karyotype of the patient showing the presence of robertsonian translocation 45,XY t(14;22). b– e, TEM micrographs of ejaculated spermatozoa. The diffuse immaturity is evident in b; spermatids and spermatozoa have uncondensed round (RN) or elliptical (EN) nuclei, frequently embedded in cytoplasmic residues (CR). Apoptotic nuclei (AN) and necrotic spermatozoa (NS) as well as binucleated germinal cells (BGC) are shown. In c and d, the mitochondrial helix (M), the accessory fibers (AF), and the fibrous sheath (FS) are normal. The axoneme is regularly assembled, but the microtubular doublets are devoid of one or both dynein arms (arrows in c and d). In e, the axonemal pattern “9⫹0” is devoid of the central doublet of microtubules (arrow). b, ⫻2,850; c, ⫻60,000; d and e, ⫻78,000.
Baccetti. Translocation 14;22 in an infertile male. Fertil Steril 2002.
FERTILITY & STERILITY威
1129
tron microscopy, as well as by FISH analysis. The sperm function was evaluated by the hamster test. We have reported that at the light microscopy level, the percentage of sperm with normal morphology was at the lower limit of the WHO range (12), while the progressive motility was very poor. The ultrastructural examination by TEM demonstrated the presence of characteristic defects typical of a status of immaturity in about 90% of the sperm cells. The DNA fragmentation, revealed by Tunel, indicated that more than 20% of the spermatozoa are degenerating by apoptotic or necrotic process. The translocation 14;22 seems to induce the presence of unusual ultrastructural sperm defects. This kind of alteration, in other cases of chromosomal deficiencies, was not observed by investigators who used only light microscopy techniques. This translocation could also have an interchromosomal effect on meiotic segregation, altering the distribution of other chromosomes (17). Nevertheless, the FISH analysis demonstrated that in this patient the frequency of diploid sperm was very high (5.78%). Because of these structural and genomic defects, the sperm fertilization ability was negative in vivo and in vitro. In conclusion, we suggest that the translocation 14;22 in this patient affects the spermatogenetic process, interfering with sperm differentiation and development. References
5. 6. 7.
8.
9. 10. 11.
12. 13.
14. 15. 16.
1. Chandley AC, Edmond P, Christie S, Gowans L, Fletcher J, Frackiewicz A, et al. Cytogenetics and fertility in man. I. Karyotype and seminal analysis. Ann Hum Genet 1975;39:231–54. 2. Chandley AC, Maclean N, Edmond P, Fletcher J, Watson GS. Cytogenetics and fertility in man. II. Testicular histology and meiosis. Ann Hum Genet 1976;40:165–76. 3. Plymate SR, Bremner WJ, Paulsen CA. The association of D-group
1130 Baccetti et al.
4.
Translocation 14;22 in an infertile man
17.
chromosomal translocations and defective spermatogenesis. Fertil Steril 1976;27:139 – 44. Leonard C, Bisson JP, David G. Male sterility associated with familial translocation heterozygosity t(8;15) (q22.p11). Arch Androl 1979;2: 269 –75. Marmor D, Taillemite JL, Van Den Akker J, Portnoi MF, le Porrier N, Joye N, et al. Semen analysis in subfertile balanced-translocation carriers. Fertil Steril 1980;34:496 –502. Wyrobek AJ, Heddle JA, Bruce WR. Chromosomal abnormalities and the morphology of mouse sperm heads. Can J Genet Cytol 1975;17(4): 675– 81. In’t Veld PA, Weber RFA, Los FJ, den Hollander N, Dhont M, Pieters MH, et al. Two cases of Robertsonian translocations in oligozoospermic males and their consequences for pregnancies induced by intracytoplasmic sperm injection. Hum. Reprod 1997;12:1642– 4. Ogawa S, Araki S, Araki Y, Ohno M, Sato I. Chromosome analysis of human spermatozoa from an oligoasthenozoospermic carrier for a 13;14 Robertsonian translocation by their injection into mouse oocytes. Hum Reprod 2000:15:1136 –9. Matsuda T, Nonomura M, Yamamoto, Hayashi K, Yoshida O. Sperm morphology in subfertile carriers of autosomal translocations. Int J Fertil 1991;36:178 – 82. Testart J, Gautier E, Brami C, Rolet F, Sedbon E, Thebault A. Intracytoplasmic sperm injection in infertile patients with structural chromosome abnormalities. Hum Reprod 1996;11:2609 –12. Guichaoua MR, Quack B, Speed RM, Noel B, Chandley AC, Luciani JM. Infertility in human males with autosomal translocations: meiotic study of a 14;22 Robertsonian translocation. Hum Genet 1990;86: 162– 6. World Health Organization. WHO laboratory manual for the examination of human semen and semen-cervical mucus interaction. 4th ed. Cambridge: Cambridge University Press, 1999. Baccetti B, Gambera L, Moretti E, Piomboni P. A quick molecular method for the simultaneous detection in spermatozoa of nuclear, acrosomal and axonemal structure by fluorescent microscopy. J Submicrosc Cytol Pathol 1999;31:563–9. Baccetti B, Bernieri G, Burrini AG, Collodel G, Crisa` N, Mirolli M, et al. Notulae seminologicae. 5. Mathematical evaluation of interdependent submicroscopic sperm alterations. J Androl 1995;16:365–71. Benet J, Navarro J, Genesca` A, Egozcue J, Templado C. Chromosome abnormalities in human sperm after albumin or TEST-Yolk capacitation. Hum Reprod 1991;6:369 –75. Kovanci E, Kovacs T, Moretti E, Vigue L, Bray-Ward P, Ward DC, et al. FISH assessment of aneuploidy frequencies in mature and immature human spermatozoa classified by the absence or presence of the cytoplasmic retention. Hum Reprod 2001;16: 1209 –17. Blanco J, Egozcue J, Vidal F. Interchromosomal effects for chromosome 21 in carriers of structural chromosome reorganizations determined by fluorescence in situ hybridization of sperm nuclei. Hum Genet 2000;106:500 –5.
Vol. 78, No. 5, November 2002
Infertile spermatozoa in a human carrier of robertsonian translocation 14;22 Baccio Baccetti, M.D.,a,b,c Serena Capitani, M.D., a,b,c Giulia Collodel, M.D.,a Mariela Estenoz, B.S.,b Laura Gambera, Ph.D.,a and Paola Piomboni, M.D.a,c University of Siena and Azienda Ospedaliera Senese (A.O.S.) Hospital, Siena, Italy
Received December 13, 2001; revised and accepted March 8, 2002. Supported by the Ministry of the University, Scientific and Technological Research and the University of Siena, MURST-COFIN 2000, and A.O.S. Hospital, Siena, Italy. Reprint requests: Baccio Baccetti, Department of Pediatrics, Obstetrics, and Reproductive Medicine, Section of Biology, via T. Pendola 62, 53100 Siena, Italy (FAX: 39-0577-233509; E-mail: [email protected]). a Department of Pediatrics, Obstetrics, and Reproductive Medicine, Section of Biology, University of Siena. b National Research Council, Center for the Study of Germinal Cells, A.O.S. Hospital. c Regional Reference Center for Male Infertility, A.O.S. Hospital. 0015-0282/02/$22.00 PII S0015-0282(02)03379-4
Objective: To present the ultrastructural, functional, and chromosomal analyses of spermatozoa from an infertile man with normal phenotype and chromosomal translocation 14;22. Design: Case report. Setting: Regional Reference Center for Male Infertility in Siena, Italy. Patient(s): A 36-year-old man with primary infertility for 3 years and his parents. Intervention(s): Family history and lymphocytic karyotypes, physical and hormonal assays, and semen analysis. Main Outcome Measure(s): Morphological sperm evaluation was performed by light, fluorescent, and electron microscopy; chromosomal constitution was examined by the fluorescence in situ hybridization (FISH) technique. The penetration ability of spermatozoa was checked by the hamster test. Result(s): The spermatozoa of the patient showed unusual ultrastructural defects. The nuclei were large, spheroidal, and generally uncondensed; the acrosomes were frequently absent or reduced; and the axonemes were often devoid of dynein arms or central singlet tubules. These characteristics are related to immaturity. The lymphocytic karyotype revealed a robertsonian translocation 14;22 in the sterile patient and his mother. FISH sperm analysis demonstrated a high frequency of diploidy for the chromosome 18,XY. The hamster penetration test gave negative results. Conclusion(s): The unusual structural sperm immaturity is associated with the translocation 14;22. This chromosomal anomaly may therefore negatively influence the spermatogenesis; an interchromosomal effect on meiosis segregation is also suggested. (Fertil Steril威 2002;78:1127–30. ©2002 by American Society for Reproductive Medicine.) Key Words: Robertsonian translocation, sperm function, FISH, human infertility, electron microscopy
A number of investigators have suggested that genetic and chromosomal factors can contribute to human male factor infertility, particularly in azoospermic and oligozoospermic men (1). This impairment of spermatogenesis in individuals with chromosomal anomalies has been demonstrated by Chandley et al. (2) and seems to be particularly due to translocations (3, 4). This view was criticized by Marmor et al. (5), who did not find any differences in sperm number, motility, and morphology in the carriers of balanced translocations when compared with individuals with normal karyotypes. In addition, Wyrobek et al. (6) previously substantiated this opinion in studies on mice. These differences of opinion are still maintained, even when the use of electron micros-
copy allows a more accurate analysis of sperm characteristics. In’t Veld et al. (7) and Ogawa et al. (8) correlate extreme oligoasthenozoospermia with the robertsonian translocations 13;14 and 13;13. Matsuda et al. (9) and Testart et al. (10), however, conclude that structural chromosomal anomalies, namely, different balanced autosomal translocations or inversions in the chromosome 2 and 14, seem to be unrelated to sperm motility and morphology. These conflicting opinions may derive from the different methods used by the investigators in assessing the sperm quality. Guichaoua et al. (11) performed a pachytene analysis in a sterile male 14;22 robertsonian translocation carrier. They reported that the patient was oligoasthenozoospermic, with normal sperm morphology at the light microscopy level. 1127
We performed semen analyses by light and electron microscopy and by the fluorescence in situ hybridization (FISH) technique in a sterile male carrier of a robertsonian translocation 14;22, and we found unusual structural and genomic sperm anomalies.
CASE REPORT A 36-year-old man presented to our laboratory for semen analysis after 3 years of sexual intercourse without conception. His wife, also 36, did not have any fertility problems. The patient was an only child as was his mother. Sexual development, medical history, physical examination, and hematic hormonal levels were normal. The patient did not have a ciliary defect. Total T, cortisol, T3, T4, TSH, E2, FSH, LH, prolactin, and inhibin levels were obtained. In 1999, this man underwent assisted fertilization twice. The first time, four oocytes were inseminated by IVF, but no activation was achieved. Seven months later, four more oocytes underwent IVF without sperm penetration. A subsequent intracytoplasmic sperm injection also had negative results. The lymphocytic karyotype of the patient (Fig. 1a) revealed the presence of a robertsonian translocation 45,XY t(14;22). The karyotype analysis of his mother showed the same translocation. Semen analysis was performed three times at 1-month intervals in 1999 and once in 2001. The samples were collected through masturbation after 4 days of sexual abstinence. The ejaculate was fully liquefied and then volume, pH, number, and motility of spermatozoa were evaluated following the World Health Organization (WHO) guidelines (12). The sperm number in the four different examinations was between 43,500,000 and 120,000,000; pH, 7.8; volume, 2.5–3 mL; and sperm progressive motility between 8% and 17%. Severe asthenospermia was found in the four samples. The eosin Y test revealed 28% live spermatozoa. To evaluate the sperm morphology by light microscopy, we used a modified Papanicolau staining, according to the protocol reported in the WHO guidelines (12). In all the samples, we observed that 30% of sperm had normal morphology and that the percentage of spermatozoa, bi- or multinucleated, was very high (27%). We further applied a trichromatic staining (13) with different molecular probes for the simultaneous detection of acrosomal content, chromatin organization, and axonemal texture. The spermatozoa, examined by fluorescent microscopy, showed a nuclear positivity to the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling (TUNEL), revealing DNA fragmentation in 21.3% of the cells; the acrosome was badly shaped or absent in 56% of the cells; the axonemal shape and presence of tubulin resulted in an alteration in 55.25% of the sperm. 1128 Baccetti et al.
Translocation 14;22 in an infertile man
For the evaluation of sperm ultrastructure, carried out on two different samples in 1999 and in 2001, we performed transmission electron microscopic (TEM) analysis on ejaculated spermatozoa as reported in Baccetti et al. (14). These observations of 300 suitable sperm sections, cut from several areas of the ejaculate, point out peculiar anomalies of the head structure, characteristic of sperm immaturity (Fig. 1b): the nuclei were irregularly shaped, often elliptical or spherical, with uncondensed or marginated chromatin (structural characteristic of apoptosis). Moreover, spermatozoa binucleate or multinucleate were frequently detected. The acrosome was absent in half of the analyzed spermatozoa; in the other half, this organelle was frequently short and badly shaped. The cytoplasmic residues were present in 50% of the spermatozoa, embedding a disorganized rolled-up axoneme, sometimes coiled around the nucleus. The mitochondrial shape and assembly frequently appeared irregular. Also, in the tails with regular shape (Figs. 1c and d), the axonemal structure was often altered, with absent central tubules (Fig. 1e) and with some doublets having incomplete or absent dynein arms. The accessory fibers and the fibrous sheath were rather well organized in about 50% of the examined tail sections. After the statistical elaboration of TEM data with the formula of Baccetti et al. (14), we found that in the examined samples the mean of the percentages of spermatozoa probably devoid of defects was ⬃0.050%, corresponding to a mean of the total numbers of “healthy spermatozoa” of ⬃50,000. These values are very low. To thoroughly investigate the penetration capacity of the spermatozoa, we performed the hamster test, according to Benet et al. (15). Zona-free hamster oocytes and sperm samples of the patient and two different donors were coincubated. The experiment was repeated twice. Penetration was observed only in the donor’s samples and never in the patient’s samples. Subsequently, we carried out FISH analysis, according to Kovanci et al. (16), to evaluate the aneuploidy frequency in ejaculated spermatozoa. A mix of probes (CEP, Vysis, IL) for chromosomes 18,XY was used. The disomy frequency of the analyzed chromosomes, evaluated in 5,474 spermatozoa, was in the standard range compared with the values found in fertile individuals. The observed frequency of diploidies for the investigated chromosomal sets was very high (5.78% of spermatozoa), despite the fact that the nuclei of these sperm after 6-diamino-2-phenylindole staining were shown to be single, with a well-defined edge.
DISCUSSION In this study, we report the case of a sterile male patient carrier of a robertsonian translocation 14;22 inherited from his mother. The spermatozoa of this patient were examined by several techniques, including light, fluorescent, and elecVol. 78, No. 5, November 2002
FIGURE 1 a, The lymphocytic karyotype of the patient showing the presence of robertsonian translocation 45,XY t(14;22). b– e, TEM micrographs of ejaculated spermatozoa. The diffuse immaturity is evident in b; spermatids and spermatozoa have uncondensed round (RN) or elliptical (EN) nuclei, frequently embedded in cytoplasmic residues (CR). Apoptotic nuclei (AN) and necrotic spermatozoa (NS) as well as binucleated germinal cells (BGC) are shown. In c and d, the mitochondrial helix (M), the accessory fibers (AF), and the fibrous sheath (FS) are normal. The axoneme is regularly assembled, but the microtubular doublets are devoid of one or both dynein arms (arrows in c and d). In e, the axonemal pattern “9⫹0” is devoid of the central doublet of microtubules (arrow). b, ⫻2,850; c, ⫻60,000; d and e, ⫻78,000.
Baccetti. Translocation 14;22 in an infertile male. Fertil Steril 2002.
FERTILITY & STERILITY威
1129
tron microscopy, as well as by FISH analysis. The sperm function was evaluated by the hamster test. We have reported that at the light microscopy level, the percentage of sperm with normal morphology was at the lower limit of the WHO range (12), while the progressive motility was very poor. The ultrastructural examination by TEM demonstrated the presence of characteristic defects typical of a status of immaturity in about 90% of the sperm cells. The DNA fragmentation, revealed by Tunel, indicated that more than 20% of the spermatozoa are degenerating by apoptotic or necrotic process. The translocation 14;22 seems to induce the presence of unusual ultrastructural sperm defects. This kind of alteration, in other cases of chromosomal deficiencies, was not observed by investigators who used only light microscopy techniques. This translocation could also have an interchromosomal effect on meiotic segregation, altering the distribution of other chromosomes (17). Nevertheless, the FISH analysis demonstrated that in this patient the frequency of diploid sperm was very high (5.78%). Because of these structural and genomic defects, the sperm fertilization ability was negative in vivo and in vitro. In conclusion, we suggest that the translocation 14;22 in this patient affects the spermatogenetic process, interfering with sperm differentiation and development. References
5. 6. 7.
8.
9. 10. 11.
12. 13.
14. 15. 16.
1. Chandley AC, Edmond P, Christie S, Gowans L, Fletcher J, Frackiewicz A, et al. Cytogenetics and fertility in man. I. Karyotype and seminal analysis. Ann Hum Genet 1975;39:231–54. 2. Chandley AC, Maclean N, Edmond P, Fletcher J, Watson GS. Cytogenetics and fertility in man. II. Testicular histology and meiosis. Ann Hum Genet 1976;40:165–76. 3. Plymate SR, Bremner WJ, Paulsen CA. The association of D-group
1130 Baccetti et al.
4.
Translocation 14;22 in an infertile man
17.
chromosomal translocations and defective spermatogenesis. Fertil Steril 1976;27:139 – 44. Leonard C, Bisson JP, David G. Male sterility associated with familial translocation heterozygosity t(8;15) (q22.p11). Arch Androl 1979;2: 269 –75. Marmor D, Taillemite JL, Van Den Akker J, Portnoi MF, le Porrier N, Joye N, et al. Semen analysis in subfertile balanced-translocation carriers. Fertil Steril 1980;34:496 –502. Wyrobek AJ, Heddle JA, Bruce WR. Chromosomal abnormalities and the morphology of mouse sperm heads. Can J Genet Cytol 1975;17(4): 675– 81. In’t Veld PA, Weber RFA, Los FJ, den Hollander N, Dhont M, Pieters MH, et al. Two cases of Robertsonian translocations in oligozoospermic males and their consequences for pregnancies induced by intracytoplasmic sperm injection. Hum. Reprod 1997;12:1642– 4. Ogawa S, Araki S, Araki Y, Ohno M, Sato I. Chromosome analysis of human spermatozoa from an oligoasthenozoospermic carrier for a 13;14 Robertsonian translocation by their injection into mouse oocytes. Hum Reprod 2000:15:1136 –9. Matsuda T, Nonomura M, Yamamoto, Hayashi K, Yoshida O. Sperm morphology in subfertile carriers of autosomal translocations. Int J Fertil 1991;36:178 – 82. Testart J, Gautier E, Brami C, Rolet F, Sedbon E, Thebault A. Intracytoplasmic sperm injection in infertile patients with structural chromosome abnormalities. Hum Reprod 1996;11:2609 –12. Guichaoua MR, Quack B, Speed RM, Noel B, Chandley AC, Luciani JM. Infertility in human males with autosomal translocations: meiotic study of a 14;22 Robertsonian translocation. Hum Genet 1990;86: 162– 6. World Health Organization. WHO laboratory manual for the examination of human semen and semen-cervical mucus interaction. 4th ed. Cambridge: Cambridge University Press, 1999. Baccetti B, Gambera L, Moretti E, Piomboni P. A quick molecular method for the simultaneous detection in spermatozoa of nuclear, acrosomal and axonemal structure by fluorescent microscopy. J Submicrosc Cytol Pathol 1999;31:563–9. Baccetti B, Bernieri G, Burrini AG, Collodel G, Crisa` N, Mirolli M, et al. Notulae seminologicae. 5. Mathematical evaluation of interdependent submicroscopic sperm alterations. J Androl 1995;16:365–71. Benet J, Navarro J, Genesca` A, Egozcue J, Templado C. Chromosome abnormalities in human sperm after albumin or TEST-Yolk capacitation. Hum Reprod 1991;6:369 –75. Kovanci E, Kovacs T, Moretti E, Vigue L, Bray-Ward P, Ward DC, et al. FISH assessment of aneuploidy frequencies in mature and immature human spermatozoa classified by the absence or presence of the cytoplasmic retention. Hum Reprod 2001;16: 1209 –17. Blanco J, Egozcue J, Vidal F. Interchromosomal effects for chromosome 21 in carriers of structural chromosome reorganizations determined by fluorescence in situ hybridization of sperm nuclei. Hum Genet 2000;106:500 –5.
Vol. 78, No. 5, November 2002