Correlation between aneuploidy, apoptotic markers and DNA fragmentation in spermatozoa from normozoospermic patients Xavier Vendrell, Minerva Ferrer,

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Presentation transcript:

Correlation between aneuploidy, apoptotic markers and DNA fragmentation in spermatozoa from normozoospermic patients Xavier Vendrell, Minerva Ferrer, Elena García-Mengual, Patricia Muñoz, Juan Carlos Triviño, Carmen Calatayud, Vanesa Y. Rawe, Miguel Ruiz-Jorro Reproductive BioMedicine Online Volume 28, Issue 4, Pages 492-502 (April 2014) DOI: 10.1016/j.rbmo.2013.12.001 Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 1 Study design. DGC, density-gradient centrifugation; ePS, externalized phosphatidylserine; Δψm, mitochondrial membrane potential; DFI, DNA fragmentation index; MACS, magnetic activated cell sorting. Reproductive BioMedicine Online 2014 28, 492-502DOI: (10.1016/j.rbmo.2013.12.001) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 2 Aneuploidy and diploidy for chromosomes X, Y and 18 in fresh, post-DGC and post-MACS fractions: (A) aneuploidy (nullisomy plus disomy) for chromosome 18; (B) aneuploidy for sex chromosomes; (C) total diploidy; (D) total aneuploidy. Horizontal lines indicate mean, boxes indicate interquartile range, whiskers indicate minimum and maximum values and asterisks indicate atypical values. DGC=density-gradient centrifugation; MACS, magnetic activated cell sorting. Reproductive BioMedicine Online 2014 28, 492-502DOI: (10.1016/j.rbmo.2013.12.001) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 3 Multichannel flow cytometry charts, showing subpopulation distributions in analysed parameters (annexin-V/PI, Δψm and DFI) in patient 1. (A) Schemes explaining the distribution of the subpopulations: grey indicates the subpopulation selected by MACS. (B) Cell distribution post-DGC. (C) Cell distribution post-MACS. Abscises and ordinates axis correspond to fluorescence intensity. DGC=density-gradient centrifugation; MACS, magnetic activated cell sorting. Reproductive BioMedicine Online 2014 28, 492-502DOI: (10.1016/j.rbmo.2013.12.001) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions

Figure 4 Postulated sequence of events in spermatogenesis. (A) Normal meiosis. (B) Meiotic defect: abnormal chromosome segregation should trigger initial apoptotic events. (C) Stage 1: the primary destabilizing phenomenon is Δψm disruption; this early damage could occur concomitantly with DNA fragmentation and the starting signals for PS externalisation; stage-1 damaged spermatozoa should not be retained by the annexin-V column and would be found in the fraction eluted after MACS. (D) Stage 2: advanced injured spermatozoa; these most-affected spermatozoa have ePS and should be retained by MACS. (E) MACS: sperm retention dependent on the suggested stages. Δψm=mitochondrial membrane potential; DFI=DNA fragmentation index; ePS=externalized phosphatidylserine; FISH=fluorescent in-situ hybridization; MACS=magnetic activated cell sorting. Reproductive BioMedicine Online 2014 28, 492-502DOI: (10.1016/j.rbmo.2013.12.001) Copyright © 2013 Reproductive Healthcare Ltd. Terms and Conditions