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

IF 1 4区生物学 Q3 BIOLOGY Cryo letters Pub Date : 2023-09-01

M Dutta, G Kadirvel, P Borah, S Sinha, K Ahmed, G Hazarika, R Sharma, H Choudhury, S Deori, M Das Gupta, R K Biswas, S Tamuly, P M Barua, J Hussain

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Abstract

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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膜稳定剂对山羊精液冷冻保存过程中精液质量及精子膜蛋白表达的影响。

背景:精液冷冻保存是一个复杂的过程，在此过程中精子和精浆蛋白的表达、蛋白分子量或膜蛋白的丢失都会发生改变。为了弥补这些变化，不同的膜稳定剂被用于冷冻精液填充剂。然而，这方面的研究在山羊精液冷冻保存方面还很缺乏。目的:探讨膜稳定剂对山羊精液冷冻保存过程中精子膜蛋白表达的影响。材料与方法:采用人工阴道法采集9只2 ~ 2.5岁阿萨姆山公山羊36枚精液。在三柠檬酸果糖蛋黄甘油(TCFEYG)填充剂中分别添加50和80 mM蔗糖、50和100 mM海藻糖、100和150 ng / mL IGF-1(胰岛素样生长因子1蛋白)3种不同浓度的膜稳定剂，冷冻保存精液样品。采用SDS-PAGE技术对新鲜和低温保存的精子进行了膜蛋白谱分析。结果:新鲜精液精膜提取物的SDS- PAGE显示有24条分子量在10 ~ 240 kDa之间的蛋白条带。添加50 mM蔗糖和80 mM蔗糖的样品显示21条分子量在10 ~ 240 kDa之间的蛋白质条带。21条蛋白带均与新鲜精子精子膜上的蛋白带相同，但冷冻精子中不存在23 kDa、29 kDa和42 kDa。同样，在50 mM海藻糖和100 mM海藻糖的冷冻精液中，发现了22条分子量在10 ~ 240 kDa之间的蛋白带，但缺乏29 kDa和42 kDa的蛋白带。在添加100 ng / mL IGF-1和150 ng / mL 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，添加海藻糖(100 mM)和IGF-1 (100 ng/mL或150 ng/mL)可以改善解冻后精液的特性，并保护某些与生育有关的精子膜蛋白。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.