Utilización de la fluorescencia de azul de anilina como nuevo método para evaluar la compactación de la cromatina en espermatozoides
2016
Chacón de Castro, A.
It is estimated that male factors related to semen quality constitute half of the infertility causes. However, current techniques used to assess semen quality are not always able to predict the sperm fertilizing capacity or the resulting embryo quality and their implantation capability and fetal development to term. Therefore, it is still currently under investigation the finding of new techniques to assess the seminal quality having greater predictive power of reproductive success. Currently the techniques that are being studied and developed are focus on assessing the state of the sperm chromatin. This is because their level of compaction in the sperm chromatin plays an important role in motility, epigenetics, DNA protection, etc. which are essential for sperm fertilizing capacity and generate a high quality embryo. One of the techniques employed to evaluate the chromatin compaction is aniline blue staining that stains residual histones present in the chromatin of immature or defective spermatozoa. In this paper we reviewed this staining in order to improve its objectivity and we propose a methodology to quantify the intensity of staining and therefore the level of chromatin compaction. We have also discovered an unknown fluorescent property of aniline blue staining that we have used to assess the chromatin compaction by fluorescence microscopy and flow cytometry. With this new methodology we were able to discern between mature and immature populations of spermatozoa in mouse, pig and human faster and more objectively than the current one. The results show a great potential of the technique for its use in medical and veterinary clinics as well as for basic research on the structure of the sperm chromatin
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