Effects of age on DNA double-strand breaks and apoptosis in human sperm (original) (raw)
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The effects of male age on sperm DNA damage: an evaluation of 2,178 semen samples
JBRA assisted reproduction, 2018
This study aimed to evaluate the effects of male age on sperm DNA damage. This cross-sectional study included semen samples collected from 2,178 men seen at an infertility clinic. For DNA integrity analysis, the proportions of spermatozoa showing DNA fragmentation (TUNEL assay), abnormal chromatin packaging/underprotamination (chromomycin A), abnormal mitochondrial membrane potential (MMP/MitoTracker Green), and apoptosis (annexin V) were recorded. For group comparisons, enrolled subjects were divided into three groups based on their ages: ≤35 years; 36-44 years; and ≥45 years. The associations between age and sperm parameters were assessed using Spearman's rank correlation coefficient. Although aging did not affect sperm apoptosis (>.05), sperm DNA fragmentation and MMP deteriorated significantly with age (<.05). Chromatin packaging/protamination improved significantly with age (<.05). Sperm DNA fragmentation worsened with age and was apparently associated with mitocho...
The effects of male age on sperm DNA damage in an infertile population
Reproductive BioMedicine Online, 2007
The objective was to investigate the influence of age on sperm DNA damage. Semen samples were collected from 508 men in an unselected group of couples attending infertility investigation and treatment. DNA fragmentation in spermatozoa was measured by TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assay; at least 200 spermatozoa in randomly selected areas of microscope slides were evaluated using a fluorescent microscope and the percentage of TUNEL positive spermatozoa was determined. The number of cells with red fluorescence (TUNEL positive) was expressed as a percentage of the total sample [DNA fragmentation index (DFI)]. Age was treated as a continuous variable for regression and correlation analysis. The following male age groups were used: Group I: ≤35 years, Group II: 36-39 years, and Group III: ≥40 years. DFI was significantly lower in Group I than in Group II (P = 0.034) or III (P = 0.022). There was no difference in DFI between Groups II and III. In addition, regression analysis demonstrated a significant increase in sperm DFI with age (P = 0.02). TUNEL assay clearly demonstrates an increase in sperm DNA damage with age.
Archivio Italiano di Urologia e Andrologia
Objective: the aim of our study was to put forward insights to treat any possible correlation among sperm quality, sperm DNA damage and male age as they may have fertility implications for men who choose to delay fatherhood. Materials and methods: Our study is a non-interventional retrospective analysis of 3124 semen samples from patients that were investigated for the conventional semen parameters. Tunel test assay was set up for the evaluation of the sperm DNA fragmentation index (DFI). We applied the Kappa index to compare both the 1999 and the 2010 World Health Organization (WHO) reference criteria to evaluate the competence of such semen parameters categorization during the standard routine of our laboratory. Results: With regards to our findings, it is possible to underline a significant relationship between aging and semen volume (p = 0.001), motility (p = 0.009), semen viscosity (p < 0.003) and sperm DNA damage (p < 0.009). We found a trend when focusing on the semen c...
The Relationship Between Human Semen Characteristics and Sperm Apoptosis: A Pilot Study
Journal of Andrology, 2006
ABSTRACT: This work was undertaken to explore the association between human semen characteristics and apoptosis in ejaculated sperm. We collected semen samples from 23 consecutive male patients who presented to the Andrology Laboratory at Massachusetts General Hospital (MGH) for routine semen analysis. Sperm concentration and motility were measured using computer‐assisted sperm analysis. Morphology was assessed using Tygerberg strict criteria. The DNA diffusion assay was used to assess the percentage of apoptosis in ejaculated sperm. In this assay, cells were mixed with agarose and placed into a microgel on a microscopic slide. The cells were stained with YOYO‐1 dye, and apoptotic cells were viewed under a fluorescent microscope. Among 23 men, the mean (SD) sperm concentration, percent motility, percent progressive motility, and normal morphology were 125.5 (92.3) million/mL, 45.6% (22.2), 28.4% (15.2), and 8.0 (4.6), respectively. The mean (SD) percent of apoptosis in ejaculated sp...
Measuring apoptosis in human spermatozoa: a biological assay for semen quality?
Fertility and Sterility, 2000
Objective: To determine whether apoptosis can be measured in ejaculated spermatozoa by flow cytometry using the Annexin V assay, which measures expression of phosphatidylserine on the outer leaflet of the cell membrane, or the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP [deoxy-uridine triphosphate] nick end labeling) assay, which measures occurrence of DNA strand breaks and [2] to correlate the outcome with routine semen variables and the hypoosmotic swelling (HOS) test. Design: Pilot study and clinical trial. Setting: Large teaching hospital and fertility center. Patient(s): Men whose semen was studied for various reasons. Main Outcome Measure(s): Percentage of apoptotic spermatozoa by two different assays, percentage of necrotic spermatozoa, concentration and motility of spermatozoa, and outcome of the HOS test. Result(s): Apoptosis can be measured in spermatozoa by flow cytometry using the Annexin V assay and the TUNEL assay. Twenty percent of spermatozoa were apoptotic according to both assays. A significant inverse correlation was seen between phosphatidylserine expression (Annexin V assay) and sperm concentration (r ϭ Ϫ0.389; PϽ.05) and motility (r ϭ Ϫ0.289; PϽ.05). A highly significant inverse correlation was seen between DNA double-strand breaks (TUNEL assay) and sperm concentration (r ϭ Ϫ0.629; PϽ.0001). Conclusion(s): Flow cytometry can easily and reliably detect phosphatidylserine expression on the outer leaflet of the cell membrane and DNA strand breaks, both of which are hallmarks of apoptosis. About 20% of ejaculated spermatozoa are apoptotic, and the concentration of spermatozoa is lower in men with more apoptotic spermatozoa. (Fertil Steril 2000;74:245-50.
Infertile men older than 40 years are at higher risk of sperm DNA damage
Reproductive biology and endocrinology : RB&E, 2014
The effect of paternal age on semen quality is controversial. In this retrospective study, the aim was to investigate the effects of advancing age on sperm parameters including reactive oxygen species (ROS), total antioxidant capacity (TAC) and sperm DNA damage in infertile men. We also examined whether paternal age >40 y is associated with higher risk of sperm DNA damage. A total of 472 infertile men presenting for infertility were divided into 4 age groups: group A: patients ≤ 30 y; group B: patients 31- 40 y, group C: ≤ 40 y and group D: patients >40 y. The following tests were performed - semen analysis according to WHO 2010 criteria, seminal ROS by chemiluminescence, TAC by colorimetric assay and sperm DNA damage by TUNEL assay - and the results were compared amongst the 4 age groups. There was no statistical difference in conventional semen parameters, TAC and ROS with advancing paternal age as well as between different age groups. However, a significant negative associa...
Fertility and Sterility, 2009
Objective: To explore the relationship between men's age and DNA damage repair proteins related to apoptosis in human testicular germ cells. Design: Retrospective case-control study. Setting: Academic institutions. Patient(s): Testicular specimens were obtained from 22 fertile volunteers aged 20-82 years. Intervention(s): Deoxyribonucleic acid repair markers were assessed using immunohistochemical staining for the cell proliferation marker [proliferating cell nuclear antigen (PCNA)]; DNA repair markers [poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1), poly(adenosine diphosphate-ribose) (PAR), X-ray repair cross-complemen-ting1(XRCC1), and apurinic/apyrimidinic endonuclease 1 (APE1)]; and apoptosis-associated markers (caspase 9, active caspase 3, and cleaved PARP-1). Main Outcome Measure(s): The prevalence and cellular localization of the above markers in testicular tissues of young, middle aged, and old men. Result(s): Statistically significant differences in DNA damage repair-associated proteins (PARP-1, PAR, XRCC1, and APE1), and apoptosis markers (caspase 9, active caspase 3, and cleaved PARP-1) were observed in testicular samples from older men. These differences were most marked in spermatocytes. Conclusion(s): The study demonstrates that there is an age-related increase in human testicular germ cell DNA break repair and apoptosis with age. (Fertil Steril Ò 2009;91:2221-9. Ó2009 by American Society for Reproductive Medicine.
The Aging Male, 2020
The effect of male aging on fertility potential is controversial and difficult to predict. The aim of our study was to determine the associations between age, basic semen parameters, and sperm DNA fragmentation (SDF). Comparison of four age-dependent groups (men 29 years, 30-35 years, 36-40 years, and >40 years) revealed a significant fall in the basic semen characteristics and sperm genomic integrity with age. Receiver operating characteristic (ROC) analysis confirmed that men >29 years had lower semen quality. In the group of men >29 years, the prevalence of men with abnormal semen parameters was higher, and these men had over a threefold higher odds ratio (OR) for abnormal semen parameters. Next, ROC analysis revealed that a threshold of 18% SDF was optimal for discriminating between men with normal and abnormal standard semen parameters. The prevalence of men with >18% SDF was higher in the group of men >29 years than in men 29 years. Older men had an almost twofold higher risk for >18% SDF than younger men. Our results suggest that age >29 years may be a causative factor of detrimental changes in semen quality, which may raise the risk for disorders of male fertility potential.
Age-related changes in human sperm DNA integrity
Aging, 2019
Abnormal standard semen characteristics and reduced sperm chromatin maturity can appear with increasing male age. However, the influence of paternal age on semen parameters is still controversial. Therefore, this study was designed to estimate the influence of paternal age not only on conventional semen characteristics but also on sperm DNA integrity. This research was carried out on ejaculated sperm cells obtained from men (n = 1124) aged ≥40 y and <40 y. Our data revealed a decreased semen volume and an increased percentage of DFI (sperm DNA fragmentation index) in older men compared to younger men in the entire study cohort, in men with normozoospermia and in men with abnormal semen parameters. Moreover, there was a higher incidence of sperm DNA damage (>10% DFI, low fertility potential) in the groups of men aged ≥40 y than in the groups of men aged <40 y. Older men had over twice the odds ratio for high sperm DNA damage as younger men. Our findings suggest a detrimental effect of advanced paternal age on sperm chromatin integrity. The data show that the evaluation of sperm DNA has greater clinical utility than standard semen analysis in case of male fertility potential assessment.