|Protamine Ratio (P1/P2) As Predictor of The Fertility Potential Of Sperm By Couple Undergoing ICSI|
|Houda Amor1,2, Ali Zeyad1, Mohamed Sobhy Bakry3, Al MandouhHussien Bosilah3, Habib Ben Ali2, Mohamed Eid Hammadeh1|
|1Department of Obstetrics & Gynaecology, Saarland University, Homburg/Saar, Germany
2Monastir University, Laboratory of Molecular Biology, Cytogenetic & Reproductive medicine, University Hospital FarhatHachad Sousse, Tunisia
3Al-Fayoum University, the Faculty of Medicine, Al-Fayoum, Egypt
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Keywords : Infertility, sperm DNA, TUNEL, CMA3, Protamine
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Objectives:To measure the protamines P1 and P2 concentrations, the standard sperm parameters and DNA fragmentation and condensation and to find out the correlations between them and the P1/P2 ratio in order to use it as a biomarker for semen quality.
Materials and Methods: 272 male semen samples were divided into three P1/P2 ratio groups: G1 (low ratio (<0.8)), G2 (normal ratio (0.8-1.2)) and G3 (high ratio (>1.2)). DNA fragmentation and condensation were evaluated by TUNEL and CMA3 assays and Protamine levels (P1 and P2) were measured using Acid Urea PAGE electrophoresis and Western-Blot.
Results: In G1, CMA3 positive and DFI (DNA fragmentation index) are negatively correlated (r=-0.371, p=0.001), P1/P2 ratio has positive correlation with DFI (r=0.652, p=0.001) but negative one with CMA3 (r=-0.623, p=0.001).
G2 demonstrated a positive correlations between sperm concentration and P1(r=0.257, p=0.011) and P2 (r=0.277, p=0.006), progressive motility and P1 (r=0.352, p=0.001). P2 correlated positively with progressive motility (r= 0.380, p<0.01), P1/P2 ratio and CMA3 correlated positively (r=0.333, p=0.001). In contrast, normal morphology showed a negative correlation (p<0.05) with P1 (r = -0.206) and P2 (r = -0.208). However, P1 has negative correlation with DFI (-0.207, p=0.041). In G3, P1 showed a negative correlation with normal morphology (r = -0.283, p=0.027), and P1/P2 ratio demonstrated negative correlation with P2 (r = -0.372, p=0.003). DFI showed positive correlation with CMA3 (r=0.299, p=0.019).
Conclusion: The P1/P2 ratio affects DNA integrity. Therefore, it plays a crucial role in human sperm quality and function and can be used asa predictor of fertility in assisted reproductive technology treatments.
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