Seminal plasma Alters surface Glycoprofile of dromedary camel cryopreserved epididymal spermatozoa
Seminal plasma Alters surface Glycoprofile of dromedary camel cryopreserved epididymal spermatozoa
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DOI：
10.1016/j.theriogenology.2021.03.008
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发表时间：
2021-03-25
期刊：
影响因子：
2.8
通讯作者：
Monaco, Davide
中科院分区：
文献类型：
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作者：
Desantis, Salvatore;Lacalandra, Giovanni Michele;Monaco, Davide
骆驼精液的高粘度仍然是其在辅助生殖技术中应用的主要障碍。附睾精(ES)暴露在精浆(SP)中可能为促进这些重要家养物种的辅助生殖技术的发展提供了一种途径。由于精子糖萼在生殖中起着关键作用，我们旨在评估SP暴露是否改变了冷冻保存的独角兽ES的表面糖基化模式。取自去的成年单峰公牛的附睾尾，逆行冲洗附睾尾，收集附睾精。收集的样品在稀释后用三柠檬酸澄清的蛋黄延长剂进行冷冻保存，同时添加和不添加15%的SP。用15种荧光凝集素分析了对照组和SP处理组精子解冻后的碳水化合物表面特征。通过计算机辅助精子分析，还研究了形态功能特性。对照精子的糖萼凝集素结合分析表明：(1)在顶体和尾部都存在以乳糖胺为末端的N-糖链(ConA、PHA-L和RCA120)。而(2)在顶体帽中观察到α-2，3-/α2，6-连接唾液酸(MALII，SNA)和以单一N-乙酰半乳糖胺残基(TN抗原)(HPA，SBA)和半乳糖B1，3N-乙酰半乳糖胺(T抗原)(PNA)终止的O-连接糖链。N-乙酰氨基葡萄糖(sWGA和GSA II)和末端半乳糖(GSA I-B-4)残基在对照精子顶体帽区也有表达。与对照组相比，SP处理的样品表现为：1)在顶体和尾部出现以乳糖胺终止的二三天线复合型N-糖链(PHA-E)，以及以TN和T抗原终止的糖链增加；2)在尾部区域含有α-2，6-连接唾液酸、N-乙酰氨基葡萄糖和无乳糖的糖链增加。对照组和精浆处理组精子胞质液滴均与ConA、PHA-E、PHA-L、RCA120、HPA、PNA、sWGA、GSA I-B-4和GSA II结合。需要进行更全面的研究，以评估SP处理的胚胎的受精能力，以便促进其在单峰骆驼辅助生殖技术中的应用。(C)2021 Elsevier Inc.保留所有权利。
The high viscosity of Camelidae semen continues to present a major impediment for its application in assisted reproduction technology. The exposure of epididymal spermatozoa (ES) to seminal plasma (SP) may provide an approach to enhance the development of assisted reproductive techniques in these important domestic species. Since the sperm glycocalyx plays a key role in reproduction we aimed to evaluate whether SP exposure modifies the surface glycosylation patterns of cryopreserved dromedary ES. Epididymal sperm was collected through retrograde flushing of the cauda epididymidis that were obtained from orchidectomized mature dromedary bulls. The collected samples were then cryopreserved after dilution with a tris citrate clarified egg yolk extender, with and without the supplementation of 15% SP. Post-thaw carbohydrate surface profiles of both control and SP-treated spermatozoa were analyzed using 15 fluorescent lectins. Morpho-functional properties were also investigated via computer assisted sperm analysis. Lectin-binding analysis of the glycocalyx in control sperm revealed the presence of (1) N-glycans terminating with lactosamine (Con A, PHA-L, and RCA(120)), in both acrosomal and tail regions. Whilst (2) alpha 2,3-/alpha 2,6-linked sialic acids (MALII, SNA), and O-linked glycans terminating with a single N-acetylgalactosamine residue (Tn antigen) (HPA, SBA) along with galactoseb1,3N-acetylgalactosamine (T antigen) (PNA) were observed in the acrosomal cap. The expression of both N-acetylglucosamine (sWGA and GSA II) and terminalagalactose (GSA I-B-4) residues was also noted in the acrosomal cap region of control sperm. Compared with controls, SP treated samples displayed: 1) the appearance of bisected ditriantennary complex-type N-glycans (PHA-E), terminating with lactosamine, as well as an increase of Oglycans terminating with Tn and T antigens in both the acrosomal and tail regions; 2) an increase in glycans containing alpha 2,6-linked sialic acid, N-acetylglucosamine, and agalactose in the tail region. The cytoplasmic droplets of both control and seminal plasma-treated sperm bound Con A, PHA-E, PHA-L, RCA(120), HPA, PNA, sWGA, GSA I-B-4, and GSA II. These results indicate that SP treatment affects the glycan composition of the dromedary camel ES glycocalyx. More comprehensive studies are required in order to evaluate the fertilization capacity of SP-treated ES in order to facilitate its application in dromedary camel assisted reproduction technology. (C) 2021 Elsevier Inc. All rights reserved.