膜稳定剂对山羊精液冷冻保存过程中精液质量和精子膜蛋白表达的影响

IF 1 4区生物学 Q3 BIOLOGY

Cryo letters Pub Date : 2023-09-01 DOI:10.54680/fr23510110612

Mitali Dutta, G. Kadirvel, Probodh Borah, S. Sinha, Kutubuddin Ahmed, Girin Hazarika, Rajeev Sharma, Hitu Choudhury, Sourabh Deori, Mohua Das Gupta, R. Biswas, S. Tamuly, Prithviraj Mazinder Barua, Jakir Hussain

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MATERIALS AND METHODS A total of 36 semen ejaculates from nine Assam Hill Goat bucks aged 2 to 2.5 years was collected by artificial vagina method. Three membrane stabilizers, each at two different concentrations viz. 50 and 80 mM sucrose, 50 and 100 mM trehalose, and 100 and 150 ng per mL IGF-1 (insulin-like growth factor 1 protein) were added to Tris-citric acid fructose egg yolk glycerol (TCFEYG) extender and semen samples were cryopreserved. The sperm membrane protein profile was studied in fresh and cryopreserved semen by SDS-PAGE. RESULTS SDS- PAGE of sperm membrane extract of fresh semen revealed the presence of 24 protein bands with molecular weights ranging from 10 kDa to 240 kDa. Samples supplemented with 50 mM sucrose and 80 mM sucrose revealed 21 protein bands with molecular weights ranging from 10 kDa to 240 kDa. 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引用次数: 0

摘要

背景精液冷冻保存是一个复杂的过程，在这一过程中，精子和精浆蛋白的表达、蛋白的分子量或膜蛋白的损失都会发生变化。为了弥补这些变化，冷冻精液扩展剂中使用了不同的膜稳定剂。然而，关于山羊精液冷冻保存的此类研究却很少。目的研究山羊精液冷冻保存过程中膜稳定剂对精子膜蛋白表达的影响。材料与方法通过人工阴道法收集了 9 只年龄在 2 至 2.5 岁之间的阿萨姆山羊的精液，共 36 份。在三柠檬酸果糖蛋黄甘油（TCFEYG）扩展剂中添加了三种膜稳定剂，分别为 50 和 80 mM 蔗糖、50 和 100 mM 曲哈糖，以及每毫升 100 和 150 ng IGF-1（胰岛素样生长因子 1 蛋白），并对精液样本进行冷冻保存。通过 SDS-PAGE 对新鲜精液和冷冻保存精液中的精子膜蛋白进行研究。结果新鲜精液精子膜提取物的 SDS- PAGE 显示存在 24 条蛋白质条带，分子量从 10 kDa 到 240 kDa 不等。添加 50 mM 蔗糖和 80 mM 蔗糖的样品显示出 21 条蛋白质条带，分子量从 10 kDa 到 240 kDa 不等。除了 23 kDa、29 kDa 和 42 kDa 蛋白条带在冷冻精液中缺失外，所有 21 条蛋白条带都与在新鲜精子的精子膜中观察到的相同。同样，用 50 mM 曲哈糖和 100 mM 曲哈糖扩增的冷冻精液显示出 22 条分子量从 10 kDa 到 240 kDa 的蛋白质条带，但缺少 29 kDa 和 42 kDa 条带。在补充了每毫升 100 毫微克 IGF-1 和每毫升 150 毫微克 IGF-1 的冷冻精液中，没有分子量为 29 kDa、130 kDa 和 240 kDa 的蛋白质。结论本研究表明，在三碱式扩展剂中添加 100 毫摩尔的三卤糖和或 100 纳克/毫升或 150 纳克/毫升的 IGF-1 可改善解冻后精液的特性，并保护某些与生育相关的精子膜蛋白。Doi.org/10.54680/fr23510110612.

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Effect of membrane stabilizers on semen quality and sperm membrane protein expression during cryopreservation of goat semen.

BACKGROUND Semen cryopreservation is a complex process during which there is alteration in the expression of sperm and seminal plasma proteins, molecular weight of protein or loss of membrane proteins during the process. In order to compensate for these changes, different membrane stabilizers are used in freezing semen extenders. However, there is scarcity of such studies during cryopreservation of goat semen. OBJECTIVE To investigate the effect of membrane stabilizers on sperm membrane protein expression during cryopreservation of goat semen. MATERIALS AND METHODS A total of 36 semen ejaculates from nine Assam Hill Goat bucks aged 2 to 2.5 years was collected by artificial vagina method. Three membrane stabilizers, each at two different concentrations viz. 50 and 80 mM sucrose, 50 and 100 mM trehalose, and 100 and 150 ng per mL IGF-1 (insulin-like growth factor 1 protein) were added to Tris-citric acid fructose egg yolk glycerol (TCFEYG) extender and semen samples were cryopreserved. The sperm membrane protein profile was studied in fresh and cryopreserved semen by SDS-PAGE. RESULTS SDS- PAGE of sperm membrane extract of fresh semen revealed the presence of 24 protein bands with molecular weights ranging from 10 kDa to 240 kDa. Samples supplemented with 50 mM sucrose and 80 mM sucrose revealed 21 protein bands with molecular weights ranging from 10 kDa to 240 kDa. All the 21 protein bands were same as those observed in the sperm membrane of fresh spermatozoa, except that the 23 kDa, 29 kDa and 42 kDa bands were absent in frozen semen. Similarly, frozen semen extended with 50 mM trehalose and 100 mM trehalose revealed 22 protein bands with molecular weights ranging from 10 kDa to 240 kDa, but lacking the 29 kDa and 42 kDa bands. Proteins with molecular weights of 29 kDa, 130 kDa and 240 kDa were absent in frozen semen supplemented with 100 ng per mL IGF-1 and 150 ng per mL IGF-1. CONCLUSION The present study revealed that supplementation of tris basic extender with trehalose at 100 mM and or IGF-1 at 100 ng/mL or 150 ng per mL improves the post-thaw semen characteristics and protects certain fertility related sperm membrane proteins. Doi.org/10.54680/fr23510110612.

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来源期刊

Cryo letters 生物-生理学

CiteScore

1.80

自引率

10.00%

发文量

审稿时长

1 months

期刊介绍： A bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.