Localisation and quantification of alkali‐labile sites in human spermatozoa by DNA breakage detection–fluorescence in situ hybridisation |
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| Authors: | E. I. Cortés‐Gutiérrez M. I. Dávila‐Rodríguez R. M. Cerda‐Flores J. L. Fernández C. López‐Fernández A. R. Aragón Tovar J. Gosálvez |
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| Affiliation: | 1. Department of Genetics, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social, IMSS, Monterrey, Mexico;2. Facultad de Enfermería, Universidad Autónoma de Nuevo León, Monterrey, Mexico;3. Genetics Unit, INIBIC, Complejo Hospitalario Universitario A Coru?a, La Coru?a, Spain;4. Laboratorio de Genética Molecular y Radiobiología, Centro Oncológico de Galicia, La Coru?a, Spain;5. Department of Urology, Unidad Medica de Alta Especialidad No. 25, IMSS, Monterrey, Mexico;6. Unit of Genetics, Department of Biology, Universidad Autónoma de Madrid, Madrid, Spain |
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| Abstract: | The localisation and quantification of constitutive alkali‐labile sites (ALSs) 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. ALSs 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. ALSs 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 ALSs, 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. |
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| Keywords: | Alkali‐labile sites DNA breakage detection
fluorescence in situ hybridisation
infertility sperm DNA structure |
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