chromomycin-a3 and Infertility

To evaluate the influence of DNA fragmentation, DNA methylation, and protamine deficiency as indicators of chromatin status on assisted reproductive technology outcome, and also to assess the relation between these parameters.. Prospective study.. Royan Institute and Isfahan Fertility and Infertility Center.. Semen samples from 92 infertile couples undergoing intracytoplasmic sperm injection (ICSI) and IVF were assessed during this study.. Semen analysis was carried out according to World Health Organization criteria. Protamine deficiency, DNA methylation, and DNA fragmentation were assessed by chromomycin A3 (CMA3), immunostaining, and sperm chromatin dispersion, respectively.. Chromomycin A3 positivity, DNA methylation, DNA fragmentation and assisted reproductive technology outcome.. Chromomycin A3 positivity shows a significant correlation with DNA fragmentation and fertilization rate. Furthermore, unlike in IVF patients, DNA fragmentation showed a significant negative correlation with fertilization rate in ICSI. A significant negative correlation was observed between DNA methylation and DNA fragmentation. In addition, no correlation was found between fertilization rate and DNA methylation in both IVF and ICSI patients.. The results reveal that in ICSI procedure DNA fragmentation, and CMA3 positivity affect the fertilization rate, whereas none of these parameters affect postfertilization development. Furthermore, both CMA3 positivity and DNA methylation affect DNA fragmentation, independently of each other. Thus, it can be concluded that these parameters may play an early role in initiation of development.

Topics: Adult; Chromatin; Chromomycin A3; DNA Fragmentation; DNA Methylation; Female; Humans; Infertility; Male; Pregnancy; Pregnancy Rate; Reproductive Techniques, Assisted; Semen Analysis; Spermatozoa; Treatment Outcome

The aim of the present work is to characterize the relationship between sperm protamine deficiency and single- and double-stranded DNA damage and to assess the diagnostic potential of chromomycin A3 (CMA3). For that purpose, semen samples from 90 human males with different clinical features were included (fertile donors, patients with recurrent pregnancy loss [RPL], and infertile patients). DNA condensation was analyzed by CMA3 and different types of DNA fragmentation were analyzed through the comet assay. A positive correlation between DNA condensation and single-stranded DNA fragmentation was found (Rs = .456; p = .05). CMA3 presented differences between fertile donors and all other groups (p < .001). Interestingly, patients with RPL, who were able to achieve a pregnancy, and infertile patients showed similar values of CMA3 (p > .05). Receiver operating characteristic curves and the profiles obtained by the combination of Comet assays and CMA3 indicate that the CMA3 test may be an interesting approach to distinguish those subjects with higher pregnancy loss risk from fertile donors (CMA3 area under the curve 0.928, with a confidence interval of 0.849-1.000). The present work shows that DNA condensation is related to oxidative damage, which affects mainly protamine-rich regions. The profiles observed in different clinical groups showed that CMA3 might be useful for the diagnosis of RPL risk when combined with Comet assays.

Topics: Abortion, Habitual; Adult; Chromatin; Chromomycin A3; Comet Assay; DNA; DNA Damage; DNA Fragmentation; DNA, Single-Stranded; Female; Fluorescent Dyes; Humans; Infertility; Male; Oxidation-Reduction; Pregnancy; Pregnancy Outcome; Protamines; ROC Curve; Sensitivity and Specificity; Spermatozoa; Varicocele

Sperm premature chromosomal condensation (PCC) has been associated with failed fertilization. Previous studies suggest that protamine deficiency or failed oocyte activation may make spermatozoa prone to PCC. However, it is not clear which of these two factors has a more profound effect on fertilization failure. In order to distinguish between these two phenomena, oocytes that failed to fertilize after intracytoplasmic sperm injection (ICSI) were artificially activated and the association between protamine deficiency and PCC was evaluated in the remaining oocytes that failed to fertilize. The results of this study reveal that after artificial activation, fertilization rate post-ICSI increased from 59.95 to 87.7% and PCC spermatozoa appeared to be present in over 50% of the remaining oocytes that failed to fertilize. The percentage of sperm PCC was significantly higher in protamine deficient samples, thus suggesting that after failed oocyte activation, sperm PCC induced by protamine deficiency may be considered as an alternative cause of failed fertilization post-ICSI. Furthermore, the results of this study did not show any correlation between pronuclei size asynchrony and protamine deficiency.

Topics: Chromomycin A3; Chromosomes; Female; Fertilization; Fertilization in Vitro; Humans; Infertility; Male; Oocytes; Protamines; Semen; Sperm Injections, Intracytoplasmic; Sperm-Ovum Interactions; Spermatozoa; Treatment Failure

We have previously postulated that the chromomycin A3 (CMA3) fluorochrome allows an indirect visualization of sperm chromatin packaging quality and partially denatured sperm DNA. In this study we investigate the relationship between CMA3 positivity and sperm morphology. We also present data on the association between sperm morphology and the presence of endogenous nicks in sperm DNA.. Semen samples were examined from 81 males of the couples who were consulting for infertility treatment. CMA3 fluorescence was assessed for all samples, while in 24 sperm samples we also examined for the presence of endogenous nicks in the sperm DNA.. When sperm morphology was less than 20% normal in a patient, the level of CMA3 fluorescence and presence of endogenous nicks were significantly higher than in patients with a higher incidence of morphologically normal sperm.. CMA3 could be used as an adjunct to the assessment of morphology as an evaluation method for poor sperm. Its value in predicting fertilizing ability when using either SUZI or ICSI awaits to be answered.

Topics: Biomarkers; Chromatin; Chromomycin A3; DNA Damage; Fertilization in Vitro; Fluorescent Dyes; Humans; Infertility; Male; Protamines; Sperm Motility; Spermatozoa