Sperm DNA fragmentation and mitochondrial membrane potential are better for predicting natural pregnancy than semen analysis

Author(s):  
Sladjana Malic
Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3309
Author(s):  
Michal Ďuračka ◽  
Kamila Husarčíková ◽  
Mikuláš Jančov ◽  
Lucia Galovičová ◽  
Miroslava Kačániová ◽  
...  

Bacterial contamination of bovine ejaculates intended for artificial insemination may be reflected in a significant economic loss due to unsuccessful fertilization as well as health issues of the recipients. The Staphylococcus genus represents a large part of bacteriocenosis of bovine ejaculates. Therefore, this study aims to get a closer look on the effects of Staphylococcus-induced bacteriospermia under in vitro conditions on bovine sperm quality. Prior to inducing bacteriospermia, spermatozoa were separated from each ejaculate using Percoll® Plus gradient medium in order to limit the effects only to the selected bacterial species. Seven Staphylococcus species previously isolated from bovine semen were used for our experiments at a turbidity of 0.5 McFarland (equivalent to 1.5 × 108 colony-forming units per mL). The contaminated semen samples were incubated at 37 °C and at times of 0, 2, and 4 h, motility, mitochondrial membrane potential, reactive oxygen species (ROS) generation, sperm DNA fragmentation, and magnesium (Mg) and calcium (Ca) extracellular concentration were analyzed and compared with the control group (uncontaminated). The results showed no significant changes at the initial measurement. However, significant adverse effects were observed after 2 h and 4 h of incubation. Most notably, the presence of S. aureus, S. warneri, S. kloosii, and S. cohnii caused a significantly increased ROS production, leading to sperm DNA fragmentation, changes in the mitochondrial membrane potential, and a decreased sperm motility. Furthermore, the presence of Staphylococcus species led to lower extracellular concentrations of Mg and Ca. In conclusion, the overgrowth of Staphylococcus bacteria in bovine semen may contribute to oxidative stress resulting in sperm DNA fragmentation, altered mitochondrial membrane potential, and diminished sperm motility.


2012 ◽  
Vol 98 (3) ◽  
pp. S47-S48
Author(s):  
L. Rubal ◽  
A.M. Hernandez ◽  
S. Ingles ◽  
M. Scrooc ◽  
K. Bendikson

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
N Kilic ◽  
T İrez ◽  
N Dayiolu

Abstract Study question Is CatSper4 expression in sperm related to functional parameters and does cryopreservation affect CatSper4 expression? Summary answer In this study, it was aimed to investigate whether CatSper4 has a relationship with sperm parameters and is CatSper 4 affected by cryopreservation. What is known already CatSper membrane channels, known as cation channels, are thought to play an important role in the insufficiency of sperm physiology, acrosome reaction, and chemotaxis movement. There is no study on cation channel distribution in an infertile male patient. In addition, studies conducted in recent years have shown that cryopreservation techniques have negative effects on sperm DNA, but there is no analysis in the literature regarding the effects of cryopreservation on CatSper4 ion channel proteins. Study design, size, duration Samples of the patients who applied to the Andrology laboratory in the Medical Park Hospital IVF unit between March 1 and June 1 in 2020 were included in the study. Also, patients with no family history of no genetic anomalies , no varicocele and azoospermia were included.The study were divided into 4 groups in accordance with the male infertility guideline of the European Association of Urology as normozoospermic (control group), the asthenoteratozoospermia, teratozoospermia, and oligoastenotheratozoospermia. Participants/materials, setting, methods In this prospective study, semen analysis, DNA fragmentation, and CatSper 4 by IHC of control group patients with normospermia (n = 40) and oligospermia(n = 50), asthenospermia(n = 40), and teratozoospermia(n = 38) patients were compared and differences resulting from cryopreservation were evaluated by Wilcoxon signed Ranks Test. Main results and the role of chance It was observed that CatSper4 protein positivity was localized in the middle part of the sperm and it was statistically higher in the normozoospermic patient group compared to the other groups (p = 0,01). When the positivity values of CatSper4 protein before and after freezing were compared in the groups, it was seen that the values decreased (p = 0,001,p=0,01). Sperm DNA fragmentation was found to be lowest in normospermia and statistically significantly higher in other groups. Cryopreservation application increased DNA fragmentation in all groups (p < 0,001 , p < 0,01). Limitations, reasons for caution Unfortunately, embryo screening in patients with low CatSper4 expression is not available in the present study. Soon we plan to screen a broader clinical pregnancy series and present the IVF results associated with CatSper4. Wider implications of the findings: Our study indicated that, CatSper4 expression is quite high in normospermia when compared with the other groups, particularly oligoasthenoteratozoospermia and asthenoteratozoospermia. There are almost no studies on this subject in the literature, and we think that it should be studied in larger patient groups and in unexplained infertile cases. Trial registration number Not applicable


2011 ◽  
pp. 166-176
Author(s):  
María Elena Maldonado-Celis ◽  
Souad Bousserouel ◽  
Francine Gossé ◽  
Annelise Lobstein ◽  
Francis Raul

Introduction: We investigated the effects of apple procyanidins (Pcy), oligomers of catechins and epicatechins on Fas receptor expression and function in human colon adenocarcinoma cells (SW480) and in their derived metastatic cells (SW620). Methods: Pcy were characterized by reverse-phase HPLC. Cell death, Fas proteins, DNA fragmentation, and mitochondrial membrane potential were analyzed by flow cytometry. Fas mRNA was analyzed by RT-PCR in real time. Results: Pcy up-regulated the expression of the Fas receptor at the cell surface of both cell lines but activated Fas gene transcription only in SW620 cells. In SW480 cells, Pcy combined with Fas agonist CH-11 enhanced Fas-mediated apoptosis involving the loss of mitochondrial membrane potential and DNA fragmentation, which were abrogated by the antagonist antibody of Fas receptor, the anti-Fas ZB4. On the contrary, in SW620 cells, CH-11 was not able to enhance Pcy-triggered apoptosis indicating that Fas receptor-mediated apoptosis was not activated in these cells despite an up-regulation of Fas receptor gene expression. However, it was observed in SW620 cells that Pcy activated the Fas receptor-mediated apoptotic pathway after a specific blockage of TRAIL-death DR4/DR5 receptors. Conclusions: The present data showed that Pcy were able to activate the Fas receptor apoptotic pathway in SW480 cells and favored a cross-talk between TRAIL and Fas receptors in SW620 cells because specific blocking of TRAIL death receptors favored activation of the Fas receptor-mediated apoptosis. These important data may allow the emergence of new therapeutic protocols targeting death receptors against resistant metastatic cells.


Zygote ◽  
2019 ◽  
Vol 28 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Thais Rose dos Santos Hamilton ◽  
Mayra Elena Ortiz D’Ávila Assumpção

SummarySperm DNA fragmentation is referred to as one of the main causes of male infertility. Failures in the protamination process, apoptosis and action of reactive oxygen species (ROS) are considered the most important causes of DNA fragmentation. Action of ROS or changes in sperm protamination would increase the susceptibility of sperm DNA to fragmentation. Routine semen analysis is unable to estimate sperm chromatin damage. Sperm DNA integrity influences sperm functional capability, therefore tests that measure sperm DNA fragmentation are important to assess fertility disorders. Actually, there is a considerable number of methods for assessing sperm DNA fragmentation and chromatin integrity, sperm chromatin stability assay (SCSA modified), sperm chromatin dispersion (SCD), comet assay, transferase dUTP nick end labelling (TUNEL); and protamine evaluation in sperm chromatin assay, such as toluidine blue, CMA3, protamine expression and evaluation of cysteine radicals. This review aims to describe the main causes of sperm DNA fragmentation and the tests commonly used to evaluate sperm DNA fragmentation.


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