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Aneuploidy and Chromosome Breakage in Swim-Up Versus Unprocessed Semen From Twenty Healthy Men
THUAN D. ONG,
LIN XUN,
SALLY D. PERREAULT,
WENDIE A. ROBBINS PhD,
THUAN D. ONG
School of Public Health, University of California, Los Angeles, Los Angeles, California
Search for more papers by this authorLIN XUN
School of Nursing, University of California, Los Angeles, Los Angeles, California
Search for more papers by this authorSALLY D. PERREAULT
US Environmental Protection Agency, Research Triangle Park, North Carolina.
Search for more papers by this authorCorresponding Author
WENDIE A. ROBBINS PhD
Center for Occupational and Environmental Health, University of California, Los Angeles, Los Angeles, California
UCLA Center for Occupational and Environmental Health, Los Angeles, CA 90095-1772 (e-mail: [email protected]).Search for more papers by this authorTHUAN D. ONG,
LIN XUN,
SALLY D. PERREAULT,
WENDIE A. ROBBINS PhD,
THUAN D. ONG
School of Public Health, University of California, Los Angeles, Los Angeles, California
Search for more papers by this authorLIN XUN
School of Nursing, University of California, Los Angeles, Los Angeles, California
Search for more papers by this authorSALLY D. PERREAULT
US Environmental Protection Agency, Research Triangle Park, North Carolina.
Search for more papers by this authorCorresponding Author
WENDIE A. ROBBINS PhD
Center for Occupational and Environmental Health, University of California, Los Angeles, Los Angeles, California
UCLA Center for Occupational and Environmental Health, Los Angeles, CA 90095-1772 (e-mail: [email protected]).Search for more papers by this authorAbstract
ABSTRACT: Toxicological and epidemiological studies have investigated several factors that are believed to induce cytogenetic damage in human sperm cells in an effort to estimate heritable risk to future generations. Most of these studies have not differentiated damage based on cell fertility or motility. In the clinical setting, intracytoplasmic sperm injection (ICSI) bypasses the natural process of sperm selection. Although practitioners attempt to select motile sperm for ICSI, the sperm may not always demonstrate motility, maturity, or even viability. Knowing whether cytogenetic damage differs in motile versus unselected sperm would improve our ability to estimate heritable risk and lead to improved ICSI procedures, and would expand the body of toxicology and epidemiology research. We divided semen samples from 20 healthy donors and compared aneuploidy and chromosome breakage in sperm cells gathered directly from the ejaculate (unprocessed semen) with cells enriched for motility using the swim-up assay. Sperm fluorescence in situ hybridization was used to detect aneuploidy for chromosomes 13, 18, 21, X, and Y. Tandem labeling probes were used to detect breakage in the 1cen-1q12 region of chromosome 1. The occurrence of disomy 1818 and XY18 was significantly lower in specimens enriched for motility (P = .004 and P = .001, respectively). Sperm that carried duplication errors and diploid sperm were also seen less frequently in semen analyzed by the swim-up assay (P < .008). Chromosome 1 breakage did not differ between swim up-assayed and unprocessed specimens. Findings suggest that unprocessed semen may overestimate heritable aneuploidy risk in sperm biomarker studies, and may be biologically relevant to ICSI in disomy categories 18–18 and XY18, demonstrating 1.4-fold to 1.8-fold differences.
References
- Baumgartner A., Van Hummelen P., Lowe XR, Adler ID, Wyrobek AJ. Numerical and structural chromosomal abnormalities detected in human sperm with a combination of multicolor FISH assays. Environ Mol Mutagen. 1999; 33: 49–58.
- Benet J., Genescá A., Navarro J., Egozcue J., Templado C. Cytogenetic studies in motile sperm from normal men. Hum Genet. 1992; 89: 176–180.
- Bernardini L., Borini A., Preti S., Conte N., Flamigni C., Capitanio GL, Venturini PL. Study of aneuploidy in normal and abnormal germ cells from semen of sterile men and infertile men. Hum Reprod. 1998; 13: 3406–3413.
- Blanco J., Rubio C., Simon C., Egozcue J., Vidal F. Increased incidence of disomic sperm nuclei in a 47, XYY male assessed by fluorescent in situ hybridization (FISH). Hum Genet. 1997; 99: 413–416.
- Brandriff BF, Gordon LA, Haendel S., Ashworth LK, Carrano AV. The chromosomal constitution of human sperm selected for motility. Fertil Steril. 1986; 46: 686–690.
- Brasch JG, Rawlins R., Tarchala S., Radwanska E. The relationship between total motile sperm count and the success of intrauterine insemination. Fertil Steril. 1994; 62: 150–154.
- Casper RF, Meriano JS, Jarvi KA, Cowan L., Lucato ML. The hypo-osmotic swelling test for selection of viable sperm for intracytoplasmic sperm injection in men with complete asthenozoospermia. Fertil Steril. 1996; 65: 972–976.
- Chandley AC, Hargreave TB. Genetic anomaly and ICSI. Hum Reprod. 1996; 11: 930–932.
- Chevret E., Rousseaux S., Monteil M., Pelletier R., Cozzi J., Séle B. Meiotic segregation of the X and Y chromosome and chromosome 1 analyzed by three-color FISH in human interphase spermatozoa. Cytogenet Cell Genetà. 1995; 71: 126–130.
- Colombero LT, Hariprashad JJ, Tsai MC, Rosenwaks Z., Palermo GD. Incidence of sperm aneuploidy in relation to semen characteristics and assisted reproductive outcome. Fertil Steril. 1999; 72: 90–96.
- Devroey P., Liu J., Nagy Z., Tournaye H., Silber SJ, Van Steirteghem AC. Normal fertilization of human oocytes after testicular sperm extraction and intracytoplasmic sperm injection. Fertil Steril. 1994; 62: 639–641.
- Finkelstein S., Mukamel E., Yavetz H., Paz G., Avivi L. Increased rate of nondisjunction in sex cells derived from low-quality semen. Hum Genet. 1998; 102: 129–137.
- Francisco RG, Newberg MT, Pang MG, et al. Determining aneuploidy and Y chromosome deletions in oligoasthenoteratozoospermic patients undergoing ICSI. J Assist Reprod Genet. 1997; 14 (Suppl): 56S.
- Gianaroli L., Munne S., Magli MC, Ferraretti AP. Preimplantation genetic diagnosis of aneuploidy and male infertility. Int J Androl. 1997; 20 (suppl 3): 31–34.
- Golan R., Shochat L., Weissenberg R., Soffer Y., Marcus Z., Oschry Y., Lewin LM. Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining. Mol Hum Reprod. 1997; 3: 47–54.
- Han TH, Ford JH, Flaherty SP, Webb GC, Matthews CD. A fluorescent in situ hybridization analysis of the chromosome constitution of ejaculated sperm in a 47, XYY male. Clin Genet. 1994; 45: 67–70.
- Härkonen K., Viitanen T., Larsen SB, Bonde JP, Lähdetie J. Aneuploidy in sperm and exposure to fungicides and lifestyle factors. ASCLE-PIOS. A European concerted action on occupational hazards to male reproductive capability. Environ Mol Mutagen. 1999; 34: 39–46.
- Harrison RF, Barrett T., Stallings R. Cytogenetic analysis is essential before ICSI. Hum Reprod. 1997; 12: 1835.
- Hassold TJ, Matsuyama A., Newlands IM, Matsuura JS, Jacobs PA, Manuel B., Tsuei J. A cytogenetic study of spontaneous abortions in Hawaii. Ann Hum Genet. 1978; 41: 443–454.
- Hassold TJ. The origin of aneuploidy in humans. Basic Life Sci. 1985; 36: 103–115.
- Hassold TJ. Chromosome abnormalities in human reproductive wastage. Trends Genet. 1986; 2: 105–110.
- Hassold TJ. Nondisjunction in the human male. Curr Top Dev Biol. 1998; 37: 383–407.
- Holt WV, Moore HD, Hillier SG. Computer-assisted measurement of sperm swimming speed in human semen: correlation of results with in vitro fertilization assays. Fertil Steril. 1985; 44: 112–119.
- Holt WV, Shenfield F., Leonard T., Hartman TD, North RD, Moore HD. The value of sperm swimming speed measurements in assessing the fertility of human frozen semen. Hum Reprod. 1989; 4: 292–297.
- Hook EB. The impact of aneuploidy upon public health: mortality and morbidity associated with human chromosome abnormalities. Basic Life Sci. 1985; 36: 7–33.
- Huang HY, Lee CL, Lai YM, Chang MY, Wang HS, Chang SY, Soong YK. The impact of the total motile sperm count on the success of intrauterine insemination with husband's spermatozoa. J Assist Reprod Genet. 1996; 13: 56–63.
- Veld PA Int, Broekmans FJ, de France HF, Pearson PL, Pieters MH, Van Kooij RJ. Intracytoplasmic sperm injection (ICSI) and chromosomally abnormal spermatozoa. Hum Reprod. 1997; 12: 752–754.
- In't Veld PA, Van Opstal D., Van den Berg C., et al. Increased incidence of cytogenetic abnormalities in chorionic villus samples from pregnancies established by in vitro fertilization and embryo transfer (IVF-ET). Prenat Diagn. 1995; 15: 975–980.
- Kahn JA, Sunde A., von During V., Sordal T., Remen A., Lippe B., Siegel J., Molne K. Formation of antisperm antibodies in women treated with fallopian tube sperm perfusion. Hum Reprod. 1993; 8: 1414–1419.
- Kusakabe H., Yamakage K., Tanaka N. Detection of neocarzinostatin-in-duced translocations in human sperm chromosomes using fluorescence in situ hybridization of chromosome 2. Mutat Res. 1996; 369: 51–58.
- Lähdetie J., Ajosenpää-Saari M., Mykkänen J. Detection of aneuploidy in human spermatozoa of normal semen donors by fluorescence in situ hybridization. Environ Health Perspect. 1996; 104 (suppl 3): 629–632.
- Lähteenmäki A., Veilahti J., Hovatta O. Intra-uterine insemination versus cyclic, low-dose prednisolone in couples with male antisperm antibodies. Hum Reprod. 1995; 10: 142–147.
- Liebaers I., Bonduelle M., Van Assche E., Devroey P., Van Steirteghem A. Sex chromosome abnormalities after intracytoplasmic sperm injection. Lancet. 1995; 346: 1095.
- Martin RH. The risk of chromosomal abnormalities following ICSI. Hum Reprod. 1996; 11: 924–925.
- Martin R. Human sperm chromosome complements in chemotherapy patients and infertile men. Chromosoma. 1998; 107: 523–527.
- Martin RH, Spriggs E., Rademaker AW. Multicolor fluorescence in situ hybridization analysis of aneuploidy and diploidy frequencies in 225,846 sperm from 10 normal men. Biol Reprod. 1996; 54: 394–398.
- Martínez-Pasarell O., Vidal F., Colls P., Nogués C., Egozcue J., Templado C. Sex chromosome aneuploidy in sperm-derived pronuclei, motile sperm and unselected sperm, scored by three-color FISH. Cytogenet Cell Genet. 1997; 78: 27–30.
- McInnes B., Rademaker A., Martin R. Donor age and the frequency of disomy for chromosomes 1, 13, 21 and structural abnormalities in human spermatozoa using multicolor fluorescence in-situ hybridization. Hum Reprod. 1998, 13; 2489–2494.
- Molina J., Castilla JA, Gil T., Hortas ML, Vergara F., Herruzo A. Influence of incubation on the chromatin condensation and nuclear stability of human spermatozoa by flow cytometry. Hum Reprod. 1995; 10: 1280–1286.
- Monteil M., Rousseaux S., Chevret E., Pelletier R. Cozzi J., Séle B. Increased aneuploid frequency in spermatozoa from a Hodgkin's disease patient after chemotherapy and radiotherapy. Cytogenet Cell Genet. 1997; 76: 134–138.
- Moosani N., Pattinson HA, Carter MD, Cox DM, Rademaker AW, Martin RH. Chromosomal analysis of sperm from men with idiopathic infertility using sperm karyotyping and fluorescence in situ hybridization. Fertil Steril. 1995; 64: 811–817.
- Morel F., Mercier S., Roux C., Clavequin MC, Bresson JL. Estimation of aneuploidy levels for 8, 15, 18, X, and Y chromosomes in 97 human sperm samples using fluorescence in situ hybridization. Fertil Steril. 1997; 67: 1134–1139.
- Newberg MT, Francisco RG, Pang MG, Brugo S., Doncel GF, Acosta AA, Hoegerman SF, Kearns WG. Cytogenetics of somatic cells and sperm from a 46, XY/45, X mosaic male with moderate oligoasthenoterato-zoospermia. Fertil Steril. 1998; 69: 146–148.
- Nishikawa N., Murakami I., Ikuta K., Suzumori K. Sex chromosomal analysis of spermatozoa from infertile men using fluorescence in situ hybridization. J Assist Reprod Genet. 2000; 17: 97–102.
- Oehninger S., Acosta R., Morshedi M., Philput C., Swanson RJ, Acosta AA. Relationship between morphology and motion characteristics of human spermatozoa in semen and in the swim-up sperm fractions. J Androl. 1990; 11: 446–452.
-
Padungtod C.,
Hassold TJ,
Millie E.,
Ryan LM,
Savitz DA,
Chrisitiani DC,
Xu X.
Sperm aneuploidy among Chinese pesticide factory workers: scoring by the FISH method.
Am J Ind Med.
1999; 36: 230–238.
10.1002/(SICI)1097-0274(199908)36:2<230::AID-AJIM2>3.0.CO;2-6 CAS PubMed Web of Science® Google Scholar
- Palermo GD, Colombero LT, Schattman GL, Davis OK, Rosenwaks Z. Evolution of pregnancies and initial follow-up of newborns delivered after intracytoplasmic sperm injection. JAMA. 1996; 276: 1893–1897.
- Pang MG, Hoegerman SF, Cuticchia AJ, Moon SY, Doncel GF, Acosta AA, Kearns WG. Detection of aneuploidy for chromosomes 4, 6, 7, 8, 9, 10, 11, 12, 13, 17, 18, 21, X and Y by fluorescence in-situ hybridization in spermatozoa from nine patients with oligoastheno-teratozoospermia undergoing intracytoplasmic sperm injection. Hum Reprod. 1999; 14: 1266–1273.
- Pfeffer J., Pang M., Hoegerman SF, Osgood CJ, Stacey MW, Mayer J., Oehninger S., Kearns WG. Aneuploidy frequencies in semen fractions from ten oligoasthenoteratozoospermic patients donating sperm for intracytoplasmic sperm injection. Fertil Steril. 1999; 72: 472–478.
- Rives N., Wust S., David B., Duchesne V., Joly G., Mace B. Fluorescence in situ hybridization with chromosome paint probes: a novel approach to assess aneuploidy in human sperm nuclei. J Assist Reprod Genet. 1999; 16: 46–51.
- Robbins WA, Baulch JE, Moore D. II, Weier HU, Blakey D., Wyrobek AJ. Three-probe fluorescence in situ hybridization to assess chromosome X, Y, and 8 aneuploidy in sperm of 14 men from two healthy groups: evidence for a paternal age effect on sperm aneuploidy. Reprod Fertil Dev. 1995; 7: 799–809.
- Robbins WA, Meistrich ML, Moore D., et al. Chemotherapy induces transient sex chromosomal and autosomal aneuploidy in human sperm. Nat Genet. 1997a; 16: 74–78.
-
Robbins WA,
Vine MF,
Truong KY,
Everson RB.
Use of fluorescence in situ hybridization (FISH) to assess effects of smoking, caffeine, and alcohol on aneuploidy load in sperm of healthy men.
Environ Mol Mutagen.
1997b; 30: 175–183.
10.1002/(SICI)1098-2280(1997)30:2<175::AID-EM10>3.0.CO;2-A CAS PubMed Web of Science® Google Scholar
- Robbins WA, Rubes J., Selevan SG, Perreault SD. Air pollution and sperm aneuploidy in healthy young men. Environ Epidemiol Toxicol. 1999; 1: 125–131.
- Rubes J., Lowe X., Moore D. II, Perreault S., Slott V., Evenson D., Selevan SG, Wyrobek AJ. Smoking cigarettes is associated with increased sperm disomy in teenage men. Fertil Steril. 1998; 70: 715–723.
- Rupa DS, Eastmond DA. Chromosomal alterations affecting the 1cen-1q12 region in buccal mucosal cells of betel quid chewers detected using multicolor fluorescence in situ hybridization. Carcinogenesis. 1997; 18: 2347–2351.
- Samura O., Miharu N., He H., Okamoto E., Ohama K. Assessment of sex chromosome ratio and aneuploidy rate in motile spermatozoa selected by three different methods. Hum Reprod. 1997; 12: 2437–2442.
- Shi Q., Martin RH. Aneuploidy in human sperm: a review of the frequency and distribution of aneuploidy, effects of donor age and lifestyle factors. Cytogenet Cell Genet. 2000a; 90: 219–226.
- Shi Q., Martin RH. Spontaneous frequencies of aneuploid and diploid sperm in 10 normal Chinese men: assessed by multicolor fluorescence in situ hybridization. Cytogenet Cell Genet. 2000b; 90: 79–83.
- Silber SJ, Van Steirteghem AC, Liu J., Nagy Z., Tournaye H., Devroey P. High fertilization and pregnancy rate after sperm injection with spermatozoa obtained from testicle biopsy. Hum Reprod. 1995; 10: 148–152.
- Spano M., Cordelli E., Leter G., Lombardo F., Lenzi A., Gandini L. Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay. Mol Hum Reprod. 1999; 5: 29–37.
- Tesarik J., Mendoza C., Testart J. Viable embryos from injection of round spermatids into oocytes. N Eng J Med. 1995; 333: 525.
- Tesarik J. Sperm or spermatid conception? Fertil Steril. 1997; 68: 214–216.
- Ushijima C., Kumasako Y., Kihaile PE, Hirotsuru K., Utsunomiya T. Analysis of chromosomal abnormalities in human spermatozoa using multi-colour fluorescence in-situ hybridization. Hum Reprod. 2000; 15: 1107–1111.
- Van Dyk Q., Lanzendorf S., Kolm P., Hodgen GD, Mahony MC. Incidence of aneuploid spermatozoa from subfertile men: selected with motility versus hemizona-bound. Hum Reprod. 2000; 15: 1529–1536.
- Vegetti W., Van Assche E., Frias A., Verheyen G., Bianchi MM, Bonduelle M., Liebaers I., Van Steirteghem A. Correlation between semen parameters and sperm aneuploidy rates investigated by fluorescence in-situ hybridization in infertile men. Hum Reprod. 2000; 15: 351–365.
- Wall MB, Marks K., Smith TA, Gearon CM, Muggleton-Harris AL. Cytogenetic and fluorescent insitu hybridization chromosomal studies on in-vitro fertilized and intracytoplasmic sperm injected failed-fertilized human oocytes. Hum Reprod. 1996; 11: 2230–2238.
- Warburton D., Stein Z., Kline J., Susser M. Chromosome abnormalities in spontaneous abortion: data from the New York City study. In: IH Porter, EB Hook, eds. Human Embryonic and Fetal Death. Washington, NY: Academic Press; 1980: 261–287.
- World Health Organization WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction. 4th ed. Washington, United Kingdom: Cambridge University Press; 1992.
- Zeyneloglu HB, Baltaci V., Ege S., Haberal A., Batioglu S. Detection of chromosomal abnormalities by fluorescent in-situ hybridization in immotile viable spermatozoa determined by hypo-osmotic sperm swelling test. Hum Reprod. 2000; 15: 853–856.
