title: Chromatin structure analysis of spermatozoa from reciprocal chromosome translocation (RCT) carriers with known meiotic segregation patterns.
abstract: The presence of reciprocal chromosome translocations (RCTs), as well as sperm chromatin disturbances, is known to exert negative influence on male fertility. The aim of this study was to identify an association between chromosome structural rearrangements in male RCT carriers and sperm seminological parameters (concentration, motility, morphology), chromatin status (fragmentation and maturity), meiotic segregation pattern and observed chromosomal hyperhaploidy. Sperm samples originated from ten male RCT carriers with reproductive failure/success. TUNEL assay (DNA fragmentation) and chromomycin A3 (CMA3)/aniline blue (AB) staining (chromatin maturity) were used to analyze sperm chromatin status while fluorescent in situ hybridization (FISH) was applied to observe meiotic segregation patterns and hyperhaploidy in spermatozoa. We found that the mean level of sperm DNA fragmentation in the RCT carrier group (18.0 ± 11.9%) was significantly higher (p=0.0006) than the mean of the control group (7.5 ± 4.3%). There was no correlation observed between sperm DNA fragmentation levels (5.6-38.0%) and the frequency of genetically normal/balanced gametes (34.3-62.4%), sperm seminological quality or revealed reproductive failure. In contrast, a correlation between the frequencies of genetically normal/balanced spermatozoa and of gametes with mature chromatin was observed (CMA3: R=0.4524, p=0.2604; AB: R=0.5238, p=0.1827). A statistically significant increase in the hyperhaploidy level of selected chromosomes in all analyzed RCT carriers was documented but was not correlated to sperm seminology or fertility status. Further evaluation and additional assays toward sperm chromatin quality assessment in RCT carriers is suggested to explain the complexity of genomic structural rearrangements and its possible relevance to reproductive success or failure.
authors.affiliation: Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479 Poznan, Poland. Electronic address: martaze@man.poznan.pl.
authors.forename: Marta
authors.initials: M
authors.lastname: Olszewska
authors.forename: Monika
authors.initials: M
authors.lastname: Fraczek
authors.forename: Nataliya
authors.initials: N
authors.lastname: Huleyuk
authors.forename: Anna
authors.initials: A
authors.lastname: Czernikiewicz
authors.forename: Ewa
authors.initials: E
authors.lastname: Wiland
authors.forename: Magdalena
authors.initials: M
authors.lastname: Boksa
authors.forename: Danuta
authors.initials: D
authors.lastname: Zastavna
authors.forename: Barbara
authors.initials: B
authors.lastname: Panasiuk
authors.forename: Alina T
authors.initials: AT
authors.lastname: Midro
authors.forename: Maciej
authors.initials: M
authors.lastname: Kurpisz
mesh_terms: D000328:Adult; D002843:Chromatin / Q000648:ultrastructure*; D014128:Chromomycin A3; D002869:Chromosome Aberrations; D020090:Chromosome Segregation; D002877:Chromosomes, Human / Q000235:genetics; D053938:DNA Fragmentation; D006238:Haploidy; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008540:Meiosis; D008875:Middle Aged; D013094:Spermatozoa / Q000648:ultrastructure*; D014178:Translocation, Genetic
chemical_list: D002843:Chromatin; D014128:Chromomycin A3
keywords: Hyperhaploidy; Meiotic segregation pattern; Reciprocal chromosomal translocation; Sperm DNA fragmentation; Sperm maturity