TY - JOUR ID - 45787 TI - The Effects of EGTA on the Quality of Fresh and Cryopreserved-Thawed Human Spermatozoa JO - Iranian Journal of Medical Sciences JA - IJMS LA - en SN - 0253-0716 AU - Ebrahimi, Bahareh AU - Keshtgar, Sara AD - Department of Physiology, Shiraz Medical School, Shiraz University of Medical Sciences, Shiraz, Iran Y1 - 2020 PY - 2020 VL - 45 IS - 3 SP - 188 EP - 198 KW - Spermatozoa KW - Cryopreservation KW - Calcium KW - Egtazic acid DO - 10.30476/ijms.2019.45787 N2 - Background: Sperm cryopreservation-thawing process has damaging effects on the structure and function of sperm, namely cryoinjury. Calcium overload has been reported as a postulated mechanism for sperm damage during the first steps after thawing. This study was designed to assess the intracellular calcium (Ca2+i) after cryopreservation and to clarify the role of a calcium chelator ethylene glycol-bis (2-aminoethyl ether)-N, N, N′, N′-tetraacetic acid (EGTA) on human sperm quality.Methods: Forty semen samples were obtained from fertile men (March 2017 to 2018). The samples were randomly divided into fresh (F) and cryopreserved-thawed (CT) groups. The F and CT samples were divided into control and 1 mM EGTA-treated groups. Sperm kinematics and membrane integrity were assessed. The reactive oxygen species (ROS) and adenosine triphosphate (ATP) were measured by luminescent methods. Ca2+i, apoptotic rate, and mitochondrial membrane potential (MMP) were evaluated using flow cytometric methods. Data were compared using SPSS software, version 16.0 by ANOVA and Kruskal-Wallis test. PResults: Cryopreservation decreased sperm motility, viability, membrane integrity, Ca2+i, MMP, and induced cell apoptosis and ROS production. EGTA could not protect the cryopreserved sperm from cryoinjury. It was found to have destructive effects on fresh sperm motility and viability (P=0.009) relative to cryopreserved sperm. ATP was reduced (P=0.02) and ROS production (P=0.0001) was increased in the EGTA-treated F and CT sperms.Conclusion: Despite Ca2+i reduction by EGTA, it had no protective effects on fresh or cryopreserved sperm. We concluded that sperm cryoinjury was not dependent on calcium overload, and it was suggested that cryoinjury was mainly related to cell membranes damage. UR - https://ijms.sums.ac.ir/article_45787.html L1 - https://ijms.sums.ac.ir/article_45787_f707f03682a0d85aaea409a8d69e8805.pdf ER -