Dose-dependent effect of melatonin on postwarming development of vitrified ovine embryos

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• 作者：Sara Succu^a ; Valeria Pasciu^a ; Maria E. Manca^a ; Sara Chelucci^a ; Laura Torres-Rovira^a ; Giovanni G. Leoni^a ; Angelo Zinellu^b ; Ciriaco Carru^b ; Salvatore Naitana^a ; Fiammetta Berlinguer^a ; ^{berling@uniss.it" class="auth_mail}
• 关键词：In vitro embryo production ; Oxidative stress ; Apoptosis ; ATP ; Total antioxidant capacity
• 刊名：Theriogenology
• 出版年：May, 2014
• 年：2014
• 卷：81
• 期：8
• 页码：1058-1066
• 全文大小：444 K

文摘

After cryopreservation, embryos become sensitive to the oxidative stress, resulting in lipid peroxidation, membrane injury, and structural destruction. The present study aimed to assess the effect of increasing concentration of melatonin during postwarming culture on embryo's ability to restore its functions after cryopreservation. In聽vitro-produced blastocysts were vitrified, warmed, and cultured in聽vitro in TCM 199 with 5 different supplementations: control (CTR): 10% fetal calf serum; bovine serum albumin (BSA): 0.04% (wt/vol) BSA; and MEL^鈭?, MEL^鈭?, MEL^鈭?: BSA plus melatonin 10^鈭?, 10^鈭?, and 10^鈭?聽M. The medium with the highest melatonin concentration had the highest trolox equivalent antioxidant capacity, whose values were comparable with those determined in plasma sampled from adult ewes (8.7 卤 2.4聽mM). The other media had lower trolox equivalent antioxidant capacity values (P < 0.01), below the range of the plasma. At the same time, embryos cultured with the highest melatonin concentration reported a lower in聽vitro viability, as evaluated by lower re-expansion and hatching rates, and lower total cell number compared with the other groups (P < 0.05). Their metabolic status was also affected, as evidenced by higher oxidative and apoptotic index and lower ATP concentration. The beneficial effects of melatonin on embryo development during postwarming culture were observed only at low concentration (10^鈭?聽M). These results suggest that melatonin at high concentration may exert some degree of toxic activity on pre-implantation embryos. Thus, the dose at which the embryos are exposed is pivotal to obtain the desiderate effect.