Abstract
Fluorescence in situ hybridization (FISH) was performed on human interphase sperm nuclei to determine the utility of this technique for aneuploidy detection. Repetitive DNA sequences specific for chromosomes 1, 12 and X were biotinylated and hybridized with mature sperm, which had been treated with cetyltrimethylammonium bromide and dithiothreitol to render them accessible to the probes. Detection of bound probe was accomplished with fluoresceinated avidin and antiavidin. For each of the chromosomes studied, chromosome number was determined by counting the fluorescent signals, representing hybridized regions, within the sperm nuclei. The frequencies for disomy, that is for nuclei containing two signals, for chromosomes 1, 12 and X were 0.06%, 0.04% and 0.03%, respectively. The congruence of these results with those determined by the cross-species hamster oocyte-human sperm assay, and the high efficiency of hybridization indicate that FISH is a sensitive and reliable tool for aneuploidy detection in human sperm.
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References
Baldini A, Rocchi M, Archidiacono N, Miller OJ, Miller DA (1990) A human alpha satellite DNA subset specific for chromosome 12. Am J Hum Genet 46:784–788
Balhorn R, Gledhill BL, Wyrobek AJ (1977) Mouse sperm chromatin proteins: quantitative isolation and partial characterization. Biochemistry 16:4074–4080
Beatty RA (1977) F-bodies as Y chromosomes markers in mature human sperm heads: a quantitative approach. Cytogenet Cell Genet 18:33–49
Bobrow M, Madan K, Pearson PL (1972) staining of some specific regions of human chromosomes, particularly the secondary constriction of no. 9. Nature New Biol 238:122–124
Brandriff BF, Gordon LA (1990) Human sperm cytogenetics and the one-cell zygote. In: Allen JW (ed) Mutation induction and heritability in mammalian germ cells (Banbury report 34) Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 183–194
Cooke HJ, Hindley J (1979) Cloning of human satellite III DNA: different components are on different chromosomes. Nucleic Acids Res 6:3177–3197
Coonen E, Pieters MHEC, Dumoulin JCM, Meyer H, Evers JLH, Ramaekers FCS, Geraedts JPM (1991) Nonisotopic in situ hybridization as a method for nondisjunction studies in human spermatozoa. Mol Reprod Dev 28:18–22
Eastmond DA, Pinkel D (1990) Detection of aneuploidy and aneuploidy-inducing agents in human lymphocytes using fluorescence in situ hybridization with chromosome-specific DNA probes. Mutat Res 234:308–318
Guttenbach M, Schmid M (1990) Determination of Y chromosome aneuploidy in human sperm nuclei by nonradioactive in situ hybridization. Am J Hum Genet 46:553–558
Guttenbach M, Schmid M (1991) Non-isotopic detection of chromosome 1 in human meiosis and demonstration of disomic sperm nuclei. Hum Genet 87:261–265
Jabs EW, Goble CA, Cutting GR (1989) Macromolecular organization of human centromeric regions reveals high-frequency, polymorphic macro DNA repeats. Proc Natl Acad Sci (USA) 86:202–206
Joseph AM, Gosden JR, Chandley AC (1984) Estimation of aneuploidy levels in human spermatozoa using chromosome specific probes and in situ hybridisation. Hum Genet 66:234–238
Martin RH (1983) A detailed method for obtaining preparations of human sperm chromosomes. Cytogenet frequency of aneuploidy among individual chromosomes in 6821 human sperm chromosomes complements. Cytogenet Cell Genet 35:253–256
Martin RH, Rademaker A (1990) The frequency of aneuploidy among individual chromosomes in 6821 human sperm chromosomes complements. Cytogenet Cell Genet 53:103–107
Martin RH, Lin CC, Balkan W, Burns K (1982) Direct chromosome analysis of human spermatozoa: preliminary results from 18 normal men. Am J Hum Genet 34:459–468
Martin RH, Ko E, Rademaker A (1991) Distribution of aneuploidy in human gametes: comparison between human sperm and oocytes. Am J Med Genet 39:321–331
Meisner LF, Chuprevich TW, Johnson CB, Inhorn SL, Carter JJ (1978) Banding of human chromosomes with caesium chloride. Lancet I:100–101
Pearson PL, Bobrow M (1970) Fluorescent staining of the Y chromosome in meiotic stages of the human male. J Reprod Fertil 22:177–179
Perreault SD, Wolff RA, Zirkin BR (1984) The role of band reduction during mammalian sperm nuclear decondensation in vivo. Dev Biol 101:160–167
Pieters MHEC, Geraedts JPM, Meyer H, Dumoulin JCM, Evers JLH, Jongbloed RJE, Nederlof PM, Flier S van der (1990) Human gametes and zygotes studies by nonradioactive in situ hybridization. Cytogenet Cell Genet 53:15–19
Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83:2934–2938
Rudak E, Jacobs PA, Yanagimachi R (1978) Direct analysis of the chromosome constitution of human spermatozoa. Nature 274:911–913
Sumner AT, Robinson JA, Evans HJ (1971) Distinguishing between X, Y and YY-bearing human spermatozoa by fluorescence and DNA content. Nature New Biol 229:231–233
Wyrobek AJ, Alhborn T, Balhorn R, Stanker L, Pinkel D (1990) Fluorescence in situ hybridization to Y chromosomes in decondensed human sperm nuclei. Mol Reprod Dev 27:200–208
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Holmes, J.M., Martin, R.H. Aneuploidy detection in human sperm nuclei using fluorescence in situ hybridization. Hum Genet 91, 20–24 (1993). https://doi.org/10.1007/BF00230216
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DOI: https://doi.org/10.1007/BF00230216