DNA fragmentation kinetics and postthaw motility of flow cytometric-sorted white-tailed deer sperm1

Abstract

This study examined DNA damage and postthaw motility of white-tailed deer sperm (n = 28) before and after sex selection and conventional sorting using MoFlo XDP SX flow cytometry. Semen samples from the same individuals were treated in 4 different ways: 1) chilled-extended sperm samples (without glycerol); 2) cryopreserved conventional samples, samples directly cryopreserved after the addition of extenders; 3) cryopreserved conventionally sorted samples, sorted samples to remove the dead sperm subpopulation; and 4) cryopreserved sex-sorted samples; sorted samples to remove the dead sperm subpopulation and separation of X- and Y-chromosome-bearing sperm. In all the cases (n = 6), conventional samples showed decreased postthaw motilities (43 ± 26%) when compared with X-sorted samples (59 ± 20%; P < 0.05) and Y-sorted samples (54 ± 20%; P > 0.05). The DNA fragmentation baseline was <5% for frozen-thawed conventional samples, but even less after sex sorting and conventional sorting: 2.4 and 1.7%, respectively. On the other hand, conventional samples showed greater (P < 0.05) DNA fragmentation than the sex-sorted sperm (n = 6) at 96 h (average of 4.8 ± 4.5% and 5.3 ± 4%, respectively). Conventionally sorted samples (n = 8) did not have greater (P > 0.05) DNA fragmentation when compared with the sex-sorted samples. Fragmentation of DNA on X-chromosome and Y-chromosome-bearing sorted sperm were not significantly different (n = 10, P > 0.05) after 96 h (2.6 ± 3.6% and 2.2 ± 0.5%, respectively). Future research should be implemented for examining the fertilizing potential of sex-sorted white-tailed deer sperm (e.g., AI fertility trials).

Types, Causes, Detection and Repair of DNA Fragmentation in Animal and Human Sperm Cells

“…One of them is the sex selection of sperm to determine the gender of the offspring with over 90% accuracy. Studies have shown that sex sorting does not affect the quality of DNA on species like deer [61] and boar [62]. However, its effect is still to be determined on bovine sperm since studies have shown that, although DNA fragmentation right after thawing is higher in conventional than in sex-sorted sperm samples, a reduced DNA longevity in sex-sorted spermatozoa was detected when the samples were incubated for 48 h [63].…”

DNA fragmentation in blue mussel (Mytilus edulis) sperm: aquaculture and fisheries implications

“…With regard to the present study, it was hypothesized that in shellfish, as in mammals (Kjelland, González‐Marín, Gosálvez, López‐Fernández, Lenz, Evans & Moreno ), fish (López‐Fernández et al . ; Gosálvez, López‐Fernández, Hermoso, Fernández & Kjelland ) and reptiles (Johnston, López‐Fernández, Arroyo, Fernández & Gosálvez ), the semen holds a in every ejaculate holds a certain proportion of sperm that does not meet the minimal requirements for fertilization.…”

Azteca Breed Horse Epididymal Sperm Evaluation: A Comparison of Head, Corpus and Cauda Sperm Quality

“…In the present study, a sample of 15 × 10 6 sperm/mL was diluted in agarose to give a final concentration of 1 × 10 6 sperm/mL, placed in racks, lysed, and stained with DAPI (SIGMA, 4'-6-diamidino Obtained 2-phenylindole) and Wright stain (Accustain, Sigma) to obtain two slides for each epididymal segment sample. Kjelland et al [28] provides a more detailed sperm sample preparation protocol using the Halomax kit (Figure 1). The DNA fragmentation analyses were conducted using a fluorescence microscope (FV10i, OYMPUS) using a magnification of 1000X with the DAPI stain, but without filters for the WRIGHT stain.…”