TEM and FISH studies in sperm from men of couples with recurrent pregnancy loss

Abstract

The role of the male partner in recurrent pregnancy loss (RPL) is not clear. In this study, semen characteristics of 22 men whose partners had experienced RPL were examined by light microscopy. Sperm morphology was analysed by transmission electron microscopy (TEM) and the data were mathematically elaborated to obtain numerical indices expressing the status of an ejaculate: the fertility index and the percentage of apoptosis, necrosis and immaturity. Sperm apoptosis and necrosis were also evaluated by annexin V/propidium iodide assay. To explore the status of meiotic segregation, fluorescence in situ hybridisation (FISH) with probes for chromosomes 18, X and Y, was applied directly on sperm nuclei. Sperm characteristics from a group of men of proven fertility were used as controls. Among the considered sperm characteristics, apoptosis (P < 0.01), 1818YY diploidy (P < 0.05) and 18YY disomy (P < 0.01) scores were significantly higher in men with RPL compared with controls. Our study showed that some patients with normal semen parameters can present a slight increase in aneuploidy compared with controls, indicating a possible involvement of sperm in some cases of RPL. Chromosomal FISH analysis and chromatin tests of sperm could be included in RPL work-ups when no other cause has been detected.

Evaluation of morphological criteria of sperm quality before in vitro fertilization and intracytoplasmic sperm injection

“…Meiotic chromosome segregation in sperm nuclei (disomy and diploidy) is investigated by fiuorescence in situ hybridisation (FISH) method using probes for chromosomes 13, 17, 18, 21, X and Y (Bielanska et al 2000, Moretti et al 2007, Collodel et al 2009b, Zhou et al 2011, Mokánszki et al 2012, Perdrix et al 2013.…”

“…Living X-and Y-chromosome bearing sperm can be sorted using Hoechst 33342 (Vigil et al 2008, Garner 2009, Karabinus 2009, Alçada-Morais et al 2013. Apoptotic sperm are differentiated from necrotic using double staining with fluorescein isothiocyanate (FITC)-labeled annexin V and Hoechst 33258 (Collodel et al 2009b, de Vantery Arrighiet al 2009). The number of viable acrosome-reacted sperm can be evaluated by staining with Hoechst 33258 and fluorescein isothiocyanate-conjugated Pisum sativum agglutinin lectin (Vigil et al 2008, Tapia et al 2011, Montjean et al 2012, Vigil et al 2012.…”

Genetic counseling for men with recurrent pregnancy loss or recurrent implantation failure due to abnormal sperm chromosomal aneuploidy

“…Aneuploidy is defined as an abnormal number of chromosomes (too few or too many) in a cell due to abnormal meiosis. Thus, for men whose partners have experienced ≥2 spontaneous abortions or IVF failures, a sperm DNA fluorescence in situ hybridization (FISH) analysis is recommended to look for male sperm aneuploidy [9][10][11][12]. In this review, we discuss the importance of genetic counseling to help couples Capsule We review the literature and discuss genetic counseling for couples with RPL or recurrent implantation failure due to increased sperm aneuploidy.…”

Y chromosome microdeletions in Mexican males of couples with idiopathic recurrent pregnancy loss

“…5 Furthermore, other sperm alterations have been described, including an altered sperm score in the hypoosmotic swelling test, 6 abnormal levels of sperm DNA fragmentation analyzed by different techniques, 5,7 the presence of non-DNA apoptotic markers 5 and morphological ultrastructural abnormalities. 8 Genetic differences also have been described in males of RPL couples when compared with normal fertile controls, and these include the presence of Y chromosome microdeletions, [9][10][11] frequency of HLA-G polymorphisms, such as HLA-G*0104 or HLA-G*0105N, 12 the presence of the HLA-C2 genotype, 13 multiple thrombophilic mutations 14 and polymorphisms in methylene tetrahydrofolate reductase gene. 15 Also, other male factors such as paternal hyperhomocysteinemia, paternal age, and paternal levels of mannan-binding protein 16 have been implicated.…”