Dimethyl tyrosine conjugated cell-penetrating peptides improved bovine blastocyst formation in vitro

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science Pub Date : 2025-11-01 Epub Date: 2025-10-06 DOI:10.1016/j.anireprosci.2025.108015

Muhammad Anzar , Kosala Rajapaksha , Jaswant Singh , François Eudes

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Abstract

In vitro embryo production and cryopreservation are assisted reproductive technologies to propagate genetically superior cattle. The objective of this study was to improve blastocyst formation in fresh and vitrified D5 (IVF day = D0) bovine morulae exposed to dimethyl tyrosine (DMT) conjugated cell-penetrating peptides (CPPs; DMT-SS31 and DMT-mTP4, antioxidants). In Experiment 1, D3 and D5 embryos were exposed to either 0.5 µM Alexa 635 labeled-SS31 or -mTP4 in CR1aa medium, at 37 °C for 30 min. Half of the morulae were examined immediately, whereas the remainder were cultured to D8 (blastocyst stage). Localization of SS31 and mTP4 were observed in D3, D5 and D8 embryos. In Experiment 2, ROS generation in the presence of SS31 or mTP4 was determined in D5 morulae using H₂FFDA marker; both CPPs reduced ROS generation in morulae (P < 0.03). In Experiment 3, fresh and vitrified morulae were treated with SS31 or mTP4 (0.5 µM each) or nothing (control) in CR1aa medium, at 37 °C for 30 min. Formation of D8 blastocysts was reduced (P < 0.001) in vitrified versus fresh embryos and both SS31 and mTP4 produced more blastocysts than control (P < 0.05). In Experiment 4, concentration-dependent effects of SS31 (0, 0.5, 1 or 2 µM) on the blastocyst formation were assessed. In fresh and vitrified morulae, there were linear relationships between SS31 concentration and blastocyst formation, with 2 µM SS31 maximizing blastocyst formation (P < 0.05). In conclusion, cell-penetrating peptides in culture medium improved blastocyst formation of fresh or vitrified bovine morulae.

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二甲基酪氨酸偶联细胞穿透肽促进体外牛囊胚形成。

体外胚胎生产和低温保存是培育优良遗传牛的辅助生殖技术。本研究的目的是改善暴露于二甲基酪氨酸（DMT）结合的细胞穿透肽（CPPs； DMT- ss31和DMT- mtp4，抗氧化剂）的新鲜和玻璃化的D5（试管婴儿日= D0）牛胚泡的形成。在实验1中，D3和D5胚胎在CR1aa培养基中分别暴露于0.5 µM Alexa 635标记的ss31或-mTP4中，37°C, 30 min。立即检查半数桑葚胚，其余桑葚胚培养至D8（囊胚期）。在D3、D5和D8胚胎中观察到SS31和mTP4的定位。实验2采用H2FFDA标记测定SS31或mTP4存在时D5样品的ROS生成；两种CPPs均能减少桑椹胚中ROS的生成(P

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来源期刊

Animal Reproduction Science 农林科学-奶制品与动物科学

CiteScore

4.50

自引率

9.10%

发文量

136

审稿时长

54 days

期刊介绍： Animal Reproduction Science publishes results from studies relating to reproduction and fertility in animals. This includes both fundamental research and applied studies, including management practices that increase our understanding of the biology and manipulation of reproduction. Manuscripts should go into depth in the mechanisms involved in the research reported, rather than a give a mere description of findings. The focus is on animals that are useful to humans including food- and fibre-producing; companion/recreational; captive; and endangered species including zoo animals, but excluding laboratory animals unless the results of the study provide new information that impacts the basic understanding of the biology or manipulation of reproduction. The journal''s scope includes the study of reproductive physiology and endocrinology, reproductive cycles, natural and artificial control of reproduction, preservation and use of gametes and embryos, pregnancy and parturition, infertility and sterility, diagnostic and therapeutic techniques. The Editorial Board of Animal Reproduction Science has decided not to publish papers in which there is an exclusive examination of the in vitro development of oocytes and embryos; however, there will be consideration of papers that include in vitro studies where the source of the oocytes and/or development of the embryos beyond the blastocyst stage is part of the experimental design.