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Theses

Infertilité masculine : fragmentation de l'ADN spermatique, ségrégation méiotique et facteurs génétiques

Abstract : Infertility affects about 15% of couples with male factor found in half of the cases. This Ph.D thesisinvestigates three causes of male infertility including chromosomal abnormality, genetic disorderand factors related to alterations in sperm DNA quality. The thesis is organized into two parts.In the first part, the chromosomal equipment and sperm DNA fragmentation in gametes of infertilemen were assessed by FISH and TUNEL. On the one hand, a high rate of aneuploid gametes andsperm DNA fragmentation were observed in four patients with gonosomal mosaicism. On the otherhand, analysis of chromosomal equipment and sperm nuclear DNA in each gamete from 13 patientswith structural chromosome abnormalities showed that unbalanced gametes have more fragmentedDNA than normal or balanced ones.In the second part, a technique for analysing the transcriptome in spermatozoa was developed onfresh and frozen semen. In fact, the combination of a discontinuous density gradient and a somaticcell lysis solution makes it possible to completely eliminate somatic cells and to recover as manysperms in the semen as possible. The XS NucleoSpin RNA kit (Macherey Nagel) was found to bemore suitable for RNA extraction than the RNA extraction kit from Qiagen. The purity of sperm RNA was verified by both RT-PCR and the Bíoanalyzer 2100. These two methods have yieldedsimilar and consistent results. The microarray analysis showed that fresh sperms do not share thesame gene expression profile than frozen ones.
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Minh Huong Nguyen. Infertilité masculine : fragmentation de l'ADN spermatique, ségrégation méiotique et facteurs génétiques. Génétique humaine. Université de Bretagne occidentale - Brest, 2015. Français. ⟨NNT : 2015BRES0034⟩. ⟨tel-02014211⟩

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