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FISH on Sperms, Spermatocytes and Oocytes

  • Maria Oliver-Bonet
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

It is well known that chromosome in situ hybridization allows the unequivocal identification of targeted human somatic chromosomes. Different fluorescence in situ hybridization (FISH) techniques have been developed over the years and, following mitotic studies, meiotic analyses have been performed using these different techniques. The application of FISH protocols to meiotic cells requires the adaptation of standard protocols to the particularities of these cells. Specific sample fixation is usually required, and in some cases samples need to go through particular pretreatments to guarantee successful FISH. The application of FISH to meiotic cytogenetic research has proven to be very useful and has provided significant information about many of the processes that take place during human gametogenesis. The protocols described in this chapter illustrate the processing of different meiotic samples as well as the FISH procedures.

Keywords

Humid Chamber Synaptonemal Complex Polar Body Meiotic Chromosome Hypotonic Solution 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was supported by Fondo Investigación Sanitaria (Madrid) (project PI 051834).

References

  1. Armstrong S, Goldman A, Speed R, Hultén M (2000) Meiotic studies of a human male carrier of the common translocation, t(11;22), suggests postzygotic selection rather than preferential 3:1 MI segregation as the cause of liveborn offspring with an unbalanced translocation. Am J Hum Genet 67:601–609CrossRefPubMedGoogle Scholar
  2. Barlow AL, Hulten MA (1998) Combined immunocytogenetic and molecular cytogenetic analysis of meiosis I oocytes from normal human females. Zygote 6:27–38CrossRefPubMedGoogle Scholar
  3. Blanco J, Egozcue J, Vidal F (1996) Incidence of chromosome 21 disomy in human spermatozoa as determined by fluorescent in-situ hybridization. Hum Reprod 11:722–726PubMedGoogle Scholar
  4. Codina-Pascual M, Campillo M, Kraus J, Speicher MR, Egozcue J, Navarro J, Benet J (2006) Crossover frequency and synaptonemal complex length: Their variability and effects on human male meiosis. Mol Hum Reprod doi:10.1093/molehr/gal007Google Scholar
  5. Durban M, Benet J, Sarquella J, Egozcue J, Navarro J (1998) Chromosome studies in first polar bodies from hamster and human oocytes. Hum Reprod 13:583–587CrossRefPubMedGoogle Scholar
  6. Durban M, Benet J, Boada M, Fernandez E, Calafell JM, Lailla JM, Sanchez-Garcia JF, Pujol A, Egozcue J, Navarro J (2001) PGD in female carriers of balanced Robertsonian and reciprocal translocations by first polar body analysis. Hum Reprod Update 7:591–602CrossRefPubMedGoogle Scholar
  7. Evans EP, Breckon G, Ford CE (1964) An air-drying method for meiotic preparations from mammalian testes. Cytogenetics 15:289–294CrossRefPubMedGoogle Scholar
  8. Goldman A, Hultén M (1993) Analysis of chiasma frequency and first meiotic segregation in a human male reciprocal translocation heterozygote, t(1;11)(p36.3;q13.1), using fluorescence in situ hybridisation. Cytogenet Cell Genet 63:16–23CrossRefPubMedGoogle Scholar
  9. Goldman AS, Hulten MA (1992) Chromosome in situ suppression hybridisation in human male meiosis. J Med Genet 29:98–102CrossRefPubMedGoogle Scholar
  10. Ko E, Rademaker A, Martin R (2001) Microwave decondensation and codenaturation: A new methodology to maximize FISH data from donors with very low concentrations of sperm. Cytogenet Cell Genet 95:143–145CrossRefPubMedGoogle Scholar
  11. Kuliev A, Verlinsky Y (2004) Meiotic and mitotic nondisjunction: Lessons from preimplantation genetic diagnosis. Hum Reprod Update 10:401–407CrossRefPubMedGoogle Scholar
  12. Lynn A, Koehler KE, Judis L, Chan ER, Cherry JP, Schwartz S, Seftel A, Hunt P, Hassold TJ (2002) Covariation of synaptonemal complex length and mammalian meiotic exchange rates. Science 296:2222–2225CrossRefPubMedGoogle Scholar
  13. Martin R, Greene C, Rademaker A, Barclay L, Ko E, Chernos J (2000) Chromosome analysis of spermatozoa extracted from testes of men with non-obstructive azoospermia. Hum Reprod 15:1121–1124CrossRefPubMedGoogle Scholar
  14. Martinez-Flores I, Cabero LL, Egozcue J, Garcia M (2003) Synaptic process in the rat (Rattus norvegicus): Influence of methodology on results. Microsc Res Tech 60:450–457CrossRefPubMedGoogle Scholar
  15. Moradkhani K, Puechberty J, Bhatt S, Lespinasse J, Vago P, Lefort G, Sarda P, Hamamah S, Pellestor F (2006) Rare Robertsonian translocations and meiotic behaviour: Sperm FISH analysis of t(13;15) and t(14;15) translocations: A case report. Hum Reprod 21:3193–3198CrossRefPubMedGoogle Scholar
  16. Munne S, Escudero T, Sandalinas M, Sable D, Cohen J (2000) Gamete segregation in female carriers of Robertsonian translocations. Cytogenet Cell Genet 90:303–308CrossRefPubMedGoogle Scholar
  17. Oliver-Bonet M, Benet J, Sun F, Navarro J, Abad C, Liehr T, Starke H, Greene C, Ko E, Martin RH (2005) Meiotic studies in two human reciprocal translocations and their association with spermatogenic failure. Hum Reprod 20:683–688CrossRefPubMedGoogle Scholar
  18. Oliver-Bonet M, Liehr T, Nietzel A, Heller A, Starke H, Claussen U, Codina-Pascual M, Pujol A, Abad C, Egozcue J, Navarro J, Benet J (2003) Karyotyping of human synaptonemal complexes by cenM-FISH. Eur J Hum Genet 11:879–883CrossRefPubMedGoogle Scholar
  19. Pacchierotti F, Adler ID, Eichenlaub-Ritter U, Mailhes JB (2007) Gender effects on the incidence of aneuploidy in mammalian germ cells. Environ Res 104:46–69CrossRefPubMedGoogle Scholar
  20. Peters AH, Plug AW, van Vugt MJ, de Boer P (1997) A drying-down technique for the spreading of mammalian meiocytes from the male and female germ line. Chromosome Res 5:66–68CrossRefPubMedGoogle Scholar
  21. Pujol A, Boiso I, Benet J, Veiga A, Durban M, Campillo M, Egozcue J, Navarro J (2003) Analysis of nine chromosome probes in first polar bodies and metaphase II oocytes for the detection of aneuploidies. Eur J Hum Genet 11:325–336CrossRefPubMedGoogle Scholar
  22. Robles P, Roig I, Garcia R, Ortega A, Egozcue J, Cabero LL, Garcia M (2007) Pairing and syn-apsis in oocytes from female fetuses with euploid and aneuploid chromosome complements. Reproduction 133:899–907CrossRefPubMedGoogle Scholar
  23. Roig I, Robles P, Garcia R, Martinez-Flores I, Cabero L, Egozcue J, Liebe B, Scherthan H, Garcia M (2005) Chromosome 18 pairing behavior in human trisomic oocytes. Presence of an extra chromosome extends bouquet stage. Reproduction 129:565–575CrossRefPubMedGoogle Scholar
  24. Scherthan H, Weich S, Schwegler H, Heyting C, Harle M, Cremer T (1996) Centromere and telomere movements during early meiotic prophase of mouse and man are associated with the onset of chromosome pairing. J Cell Biol 134:1109–1125CrossRefPubMedGoogle Scholar
  25. Smith JL, Garry VF, Rademaker AW, Martin RH (2004) Human sperm aneuploidy after exposure to pesticides. Mol Reprod Dev 67:353–359CrossRefPubMedGoogle Scholar
  26. Vidal F, Moragas M, Catala V, Torello M, Santalo J, Calderon G, Gimenez C, Barri P, Egozcue J, Veiga A (1993) Sephadex filtration and human serum albumin gradients do no select spermatozoa by sex chromosome: A fluorescent in-situ hybridization study. Hum Reprod 8:1740–1743PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2009

Authors and Affiliations

  • Maria Oliver-Bonet
    • 1
  1. 1.Unitat de Biologia Cellular i Genética Médica, Facultat de Medicina, Edifici MUniversitat Autönoma de BarcelonaBellaterraSpain

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