Efficient combined FISH and PRINS technique for detection of DAZ microdeletion in human sperm
- 187 Downloads
Intracytoplasmic sperm injection (ICSI) now offers an effective therapeutic option for men with male infertility and is believed to allow transmission of genetically determined infertility to the male offspring. Transmission of DAZ (Deleted in Azoospermia) microdeletion is one of the major concerns for oligo and severe oligozoospermia patients. Screening of the Y chromosome microdeletion in the diagnostic work-up of infertile men is mainly done using polymerase chain reaction (PCR) on blood leukocytes. However, there are evidences showing that presence of DAZ in somatic cells might not be indicative of its presence in germ cell lineage. In this report we are going to describe a combined Primed in situ labeling (PRINS) and fluorescence in situ hybridization (FISH) technique to show the localization of DAZ gene as well as Y chromosome centromere on sperm nuclei. PRINS is a combination of FISH and in situ polymerization provides another approach for in situ chromosomal detection. In the present study the PRINS primers specific for DAZ genes and traditional direct labeled centromere FISH probes for Y and X chromosomes were used in order to simultaneously detect DAZ genes and sex chromosome aneuploidy in sperm samples.
KeywordsHuman sperm Y chromosome DAZ PRINS FISH
This research was supported by the Research Department of the Faculty of Medical Sciences, Tarbiat Modares University. The authors would like to express their thanks to Mrs Z. Rezaeian for arrangements for sample collection.
Conflict of Interest statement
The authors declare that there are no conflicts of interest.
- 6.Ma K, Inglis JD, Sharkey A, Bickmore WA, Hill RE, Prosser EJ, Speed RM, Thomson EJ, Jobling M, Taylor K, et al. A Y chromosome gene family with RNA-binding protein homology: candidates for the azoospermia factor AZF controlling human spermatogenesis. Cell. 1993;75:1287–95.PubMedCrossRefGoogle Scholar
- 8.Vogt PH, Edelmann A, Kirsch S, Henegariu O, Hirschmann P, Kiesewetter F, Kohn FM, Schill WB, Farah S, Ramos C, Hartmann M, Hartschuh W, Meschede D, Behre HM, Castel A, Nieschlag E, Weidner W, Grone HJ, Jung A, Engel W, Haidl G. Human Y chromosome azoospermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet. 1996;5:933–43.PubMedCrossRefGoogle Scholar
- 12.Vogt PH, Fernandes S: Polymorphic DAZ gene family in polymorphic structure of AZFc locus: Artwork or functional for human spermatogenesis? Acta Pathologica, Microbiologica et Immunologica Scandinavica 2003; 111 (online version).Google Scholar
- 18.Simoni M, Gromoll J, Dworniczak B, Rolf C, Abshagen K, et al. Screening for deletions of the Y chromosome involving the DAZ (Deleted in AZoospermia) gene in azoospermia and severe oligozoospermia. Fertil Steril. 2010;67:1753–6.Google Scholar
- 38.Pellestor F. What PRINS can do for you. Medical Science. 1998;14:935–8.Google Scholar
- 39.World Health Organization: WHO laboratory manual for the Examination and processing of human semen. Fifth edition. WHO Press, 2010.Google Scholar