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Localisation and quantification of alkali‐labile sites in human spermatozoa by DNA breakage detection–fluorescence in situ hybridisation
Author(s) -
CortésGutiérrez E. I.,
DávilaRodríguez M. I.,
CerdaFlores R. M.,
Fernández J. L.,
LópezFernández C.,
Aragón Tovar A. R.,
Gosálvez J.
Publication year - 2015
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.12250
Subject(s) - semen , comet assay , male infertility , sperm , fluorescence in situ hybridization , andrology , infertility , biology , dna , gel electrophoresis , microbiology and biotechnology , chemistry , dna damage , genetics , medicine , gene , chromosome , pregnancy
Summary The localisation and quantification of constitutive alkali‐labile sites ( ALS s) were investigated using a protocol of DNA breakage detection plus fluorescence in situ hybridisation ( DBD – FISH ) and alkaline single‐cell gel electrophoresis ( SCGE or comet assay), in spermatozoa of infertile and fertile men. Semen samples from 10 normozoospermic patients undergoing infertility treatment and 10 fertile men were included in this study. ALS s were localised and quantified by DBD – FISH . The region most sensitive to alkali treatment in human spermatozoa was located in the basal region of the head. ALS s were more frequent in spermatozoa of infertile men than in those of fertile men. These results were confirmed by SCGE comet assays. In conclusion, the most intense localisation of hybridisation signals in human spermatozoa, representing the highest density of constitutive ALS s, was not randomly distributed and was predominantly located in the base of the head. Moreover, infertile men presented with an increase in ALS frequency. Further studies are necessary to determine the association between ALS , sperm chromatin organisation and infertility.