Single-center thorough evaluation and targeted treatment of globozoospermic men

Abstract Purpose To characterize, by specific biomarkers and nucleic acid sequencing, the structural and genomic sperm characteristics of partial (PG) and complete globozoospermic (CG) men in order to identify the best reproductive treatment. Methods We assessed spermatozoa from 14 consenting men ultrastructurally, as well as for histone content, sperm chromatin integrity, and sperm aneuploidy. Additional genomic, transcriptomic, and proteomic evaluations were carried out to further characterize the CG cohort. The presence of oocyte-activating sperm cytosolic factor (OASCF) was measured by a phospholipase C zeta (PLCζ) immunofluorescence assay. Couples were treated in subsequent cycles either by conventional ICSI or by ICSI with assisted gamete treatment (AGT) using calcium ionophore (Ionomycin, 19657, Sigma-Aldrich, Saint Louis, MO, USA). Results Ultrastructural assessment confirmed complete acrosome deficiency in all spermatozoa from CG men. Histone content, sperm chromatin integrity, and sperm aneuploidy did not differ significantly between the PG (n = 4) and CG (n = 10) cohorts. PLCζ assessment indicated a positive presence of OASCF in 4 PG couples, who underwent subsequent ICSI cycles that yielded a 36.1% (43/119) fertilization with a 50% (2/4) clinical pregnancy and delivery rate. PLCζ assessment failed to detect OASCF for 8 CG patients who underwent 9 subsequent ICSI cycles with AGT, yielding a remarkable improvement of fertilization (39/97; 40.2%) (P = 0.00001). Embryo implantation (6/21; 28.6%) and clinical pregnancies (5/7; 71.4%) were also enhanced, resulting in 4 deliveries. Gene mutations (DPY19L2, SPATA16, PICK1) were identified in spermatozoa from CG patients. Additionally, CG patients unable to sustain a term pregnancy had gene mutations involved in zygote development (NLRP5) and postnatal development (BSX). CG patients who successfully sustained a pregnancy had a mutation (PIWIL1) related to sperm phenotype. PLCZ1 was both mutated and underexpressed in these CG patients, regardless of reproductive outcome. Conclusions Sperm bioassays and genomic studies can be used to characterize this gamete’s capacity to support embryonic development and to tailor treatments maximizing reproductive outcome.

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Sperm FISH and chromatin integrity in spermatozoa from a t(6;10;11) carrier

Reproduction ◽

10.1530/rep-13-0533 ◽

2014 ◽

Vol 147 (5) ◽

pp. 659-670 ◽

Cited By ~ 5

Author(s):

Marta Olszewska ◽

Nataliya Huleyuk ◽

Monika Fraczek ◽

Danuta Zastavna ◽

Ewa Wiland ◽

...

Keyword(s):

Blue Staining ◽

Sperm Chromatin ◽

Control Group ◽

Sperm Dna Fragmentation ◽

Sperm Aneuploidy ◽

Sperm Dna ◽

Sperm Fish ◽

Chromatin Integrity ◽

Meiotic Segregation Pattern ◽

High Level

Complex chromosome rearrangements (CCRs) are structurally balanced or unbalanced aberrations involving more than two breakpoints on two or more chromosomes. CCRs can be a potential reason for genomic imbalance in gametes, which leads to a drastic reduction in fertility. In this study, the meiotic segregation pattern, aneuploidy of seven chromosomes uninvolved in the CCR and chromatin integrity were analysed in the ejaculated spermatozoa of a 46,XY,t(6;10;11)(q25.1;q24.3;q23.1)mat carrier with asthenozoospermia and a lack of conception. The frequency of genetically unbalanced spermatozoa was 78.8% with a prevalence of 4:2 segregants of 38.2%, while the prevalence of the adjacent 3:3 mode was 35.3%. Analysis of the aneuploidy of chromosomes 13, 15, 18, 21, 22, X and Y revealed an approximately fivefold increased level in comparison with that of the control group, indicating the presence of an interchromosomal effect. Sperm chromatin integrity status was evaluated using chromomycin A3 and aniline blue staining (deprotamination), acridine orange test and TUNEL assay (sperm DNA fragmentation). No differences were found when comparisons were made with a control group. We suggest that the accumulation of genetically unbalanced spermatozoa, significantly increased sperm aneuploidy level and decreased sperm motility (20%, progressive) were not responsible for the observed lack of reproductive success in the analysed infertile t(6;10;11) carrier. Interestingly, in the case described herein, a high level of sperm chromosomal imbalance appears not to be linked to sperm chromatin integrity status.

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Association of gene polymorphisms of folate metabolism with sperm aneuploidy in men with low reproductive function

Visnik ukrains'kogo tovaristva genetikiv i selekcioneriv ◽

10.7124/visnyk.utgis.14.1.547 ◽

2016 ◽

Vol 14 (1) ◽

pp. 72-78

Author(s):

O. M. Fedota ◽

O. M. Feskov ◽

I. S. Zhylkova

Keyword(s):

Dna Fragmentation ◽

Embryo Implantation ◽

Reproductive Function ◽

Mthfr C677t ◽

Mthfr Gene ◽

Folate Metabolism ◽

Sperm Chromatin ◽

Sperm Dna Fragmentation ◽

Sperm Aneuploidy ◽

Polymorphic Variants

Aim. The association of polymorphic variants of genes MTHFR (C677T, A1298C) and MTRR (A66G) with sperm aneuploidy in men with low reproductive function was investigated. Methods. SNPs determinations were performed by the real-time PCR technique. Sperm DNA fragmentation analysis was performed using the method of sperm chromatin dispersion. To detect aneuploidy in spermatozoa nuclei the method of fluorescent in situ hybridization (FISH) was used. Results. Polymorphic alleles in genes of folate metabolism are associated with sperm aneuploidy in men with low reproductive function. The link between the number of alternative alleles of polymorphic variants A1298C of MTHFR gene in genotype and the average level of aneuploidy in sperm chromosome 16 is proved.Conclusion. Aneuploid sperm is able to fertilize the oocytes, but the further formation of the blastocyst and embryo implantation may be blocked at various stages of development. Understanding the genetic basis of aneuploidy in sperm of men could reduce the reproductive losses in IVF practice.Keywords: DNA fragmentation, sperm aneuploidy, MTHFR, MTRR, reproductive function.

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