Abstract
Purpose: A DNA disc chip assay based on comparative genomic hybridization (CGH) was developed to measure sperm DNA integrity. The objective was to correlate DNA integrity of heat-treated sperm with the sperm capacitation index (CI) determined from the sperm penetration assay. Methods: Basic semen and kinematic parameters were measured (N = 6). Sperm were washed in two-layer colloid suspensions and split portions incubated at either 37°C (control) or 40°C for 4 h. Single-stranded DNA of heated sperm were stained in SYBR Gold and hybridized to bisbenzimide (Hoechst 33342) stained control DNA in a membrane disc. Fluorescent intensities of the discs were measured and correlation analyses with sperm parameters performed. Results: Sperm CI was positively correlated (R = 0.737) with sperm DNA integrity. Two populations of sperm could be discerned: low capacitating sperm that initiated apoptosis and high capacitating sperm unaffected by heat shock treatment. The remaining parameters were not related to sperm DNA stability. Conclusions: Fragile DNA were found in a population of sperm associated with poor capacitation characteristics and apoptosis was observed after heat treatment. The results suggested that sperm dysfunction might be due to apoptotic sperm DNA resulting from an elevated temperature in the surroundings. The data suggested that the second population of high capacitating sperm induced chaperones such as heat shock proteins hsp 70 to protect against apoptosis.
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REFERENCES
Kallioniemi A, Kallioniemi O.-P, Sudar D, Rutovitz D, Gray JW, Waldman F, Pinkel D: Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science 1992;258:818–821
Voullaire L, Slater H, Williamson R, Wilton L: Chromosomal analysis of blastomeres from human embryos by using comparative genomic hybridization. Hum Genet 2000;106:210–217
Lichter P, Joos S, Bentz M, Lampel S: Comparative genomic hybridization: Uses and limitations. Semin Hematol 2000;37:348–357
Schena M, Shalon D, Davis RW, Brown PO: Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 1995;270:467–470
Freeman WM, Robertson DJ, Vrana KE: Fundamentals of DNA hybridization arrays for gene expression analysis. Biotechniques 2000;29:1042–1055
Kruger TF, Acosta AA, Simmons KF, Swanson RJ, Matta JF, Oehninger S: Predictive value of abnormal sperm morphology in in vitro fertilization. Fertil Steril 1988;49:112–117
Chan PJ, Corselli JU, Jacobson JD, Patton WC, King A: Spermac stain analysis of human sperm acrosomes. Fertil Steril 1999;72:124–128
Eliasson R, Treschl L: Supravital staining of human spermatozoa. Fertil Steril 1971;22:134–137
Yanagimachi R, Yanagimachi H, Rogers BJ: The use of zona-free animal ova as a test-system for the assessment of the fertilizing capacity of human spermatozoa. Biol Reprod 1976;15:471–476
Chan PJ, Corselli JU, Patton WC, Jacobson JD, King A: Heat-induced hyperactivation. J Assist Reprod Genet 1998; 15:32–38
McClure RD, Nunes L, Tom R: Semen manipulation: Improved sperm recovery and function with a two-layer Percoll gradient. Fertil Steril 1989;51:874–877
Falk RM, Silverberg KM, Fetterolf PM, Kirchner FK, Rogers BJ: Establishment of TEST-yolk buffer enhanced sperm penetration assay limits for fertile males. Fertil Steril 1990;54:121–126
Suzuki F, Yanagimachi R: Changes in the distribution of intramembranous particles and filipin-reactive membrane sterols during in vitro capacitation of golden hamster spermatozoa. Gamete Res 1989;23:335–347
Naz RK, Ahmad K, Kumar R: Role of membrane phosphotyrosine proteins in human spermatozoal function. J Cell Sci 1991;99:157–165
Honig SC, Thompson S, Lipshultz LI: Reassessment of malefactor infertility, including the varicocele, sperm penetration assay, semen analysis, and in vitro fertilization. Curr Opin Obstet Gynecol 1993;5:245–251
Lopes S, Sun JG, Jurisicova A, Meriano J, Casper RF: Sperm deoxyribonucleic acid fragmentation is increased in poorquality semen samples and correlates with failed fertilization in intracytoplasmic sperm injection. Fertil Steril 1998;69:528–532
Peschka B, Leygraaf J, Van der Ven K, Montag M, Schartmann B, Schubert R, van der Ven H, Schwanitz G: Type and frequency of chromosome aberrations in 781 couples undergoing intracytoplasmic sperm injection. Hum Reprod 1999;14:2257–2263
Alvarez JG, Minaretzis D, Barrett CB, Mortola JF, Thompson IE: The sperm stress test: A novel test that predicts pregnancy in assisted reproductive technologies. Fertil Steril 1996;65:400–405
Ravagnan L, Gurbuxani S, Susin SA, Maisse C, Daugas E, Zamzami N, Mak T, Jaattela M, Penninger JM, Garrido C, Kroemer G: Heat-shock protein 70 antagonizes apoptosisinducing factor. Nat Cell Biol 2001;3:839–843
Samali A, Robertson JD, Peterson E, Manero F, van Zeijl L, Paul C, Cotgreave IA, Arrigo AP, Orrenius S: Hsp27 protects mitochondria of thermotolerant cells against apoptotic stimuli. Cell Stress Chaperones 2001;6:49–58
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Mann, S.L., Patton, W.C., King, A. et al. Comparative Genomic Hybridization Analysis of Sperm DNA Apoptosis After Exposure to Heat Shock. J Assist Reprod Genet 19, 195–200 (2002). https://doi.org/10.1023/A:1014846113170
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DOI: https://doi.org/10.1023/A:1014846113170