Effect of isolation protocols and cryoprotectants on freezing of stallion epididymal spermatozoa.

IF 1 4区 生物学 Q3 BIOLOGY
T. Talluri, D. Jhamb, N. Paul, J. Singh, R. Dedar, S. C. Mehta, R. Legha, Y. Pal
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引用次数: 0

Abstract

BACKGROUND The recovery of spermatozoa from the cauda epididymis may be the only option to obtain genetic material from elite stallions that had undergone castration or sudden death due to colic or severe injury. OBJECTIVE To evaluate two different protocols for retrieval of stallion epididymal spermatozoa and to evaluate different cryoprotectants on the freezability of the epididymal spermatozoa. MATERIALS AND METHODS Six epididymides from three stallions were collected immediately after routine castration under general anesthesia. In the first experiment, each epididymis (of two testes) of the same stallion were processed using different methods for retrieval of the epididymal spermatozoa and were pooled and cryopreserved either using 5% glycerol or 5% dimethyl formamide (DMF) as cryoprotectant. The semen quality parameters viz., progressive motility, HOST, viability and acrosome integrity were evaluated at the fresh, pre-freeze and post-thaw stages. RESULTS Retrograde method of flushing of epididymis yielded significantly (p < 0.05) higher concentration of the stallion sperm than that of the floating method. The qualitative semen parameters i.e., viability, plasma membrane integrity and acrosome integrity were found to be significantly restored using 5% DMF as cryoprotectant in comparison to when 5% glycerol was used. CONCLUSION Retrograde flushing method of epididymis yielded significantly higher sperm concentration to that of the floating method, and 5% DMF as cryoprotectant provided acceptable freezability of stallion epididymal spermatozoa. DOI: 10.54680/fr23310110312.
分离方案和冷冻保护剂对种马附睾精子冷冻的影响。
背景从附睾尾提取精子可能是从被阉割或因绞痛或严重损伤而猝死的优秀种马获得遗传物质的唯一选择。目的评价两种不同的提取种马附睾精子的方法,并评价不同的冷冻保护剂对附睾精子冷冻性的影响。材料与方法在全身麻醉下常规去势后立即采集3匹公马的6块附睾。在第一个实验中,对同一种马的两个睾丸的每个附睾采用不同的方法提取附睾精子,并使用5%甘油或5%二甲基甲酰胺(DMF)作为冷冻保护剂进行低温保存。在新鲜、冷冻前和解冻后阶段对精液质量参数,即进行活力、寄主、活力和顶体完整性进行了评估。结果逆行法冲洗附睾可显著提高雄马精子浓度(p < 0.05)。与使用5%甘油相比,使用5% DMF作为冷冻保护剂可显著恢复精液的定性参数,即活力、质膜完整性和顶体完整性。结论附睾逆行冲洗法精子浓度明显高于浮法,5% DMF作为冷冻保护剂可使种马附睾精子具有较好的冷冻性。DOI: 10.54680 / fr23310110312。
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来源期刊
Cryo letters
Cryo letters 生物-生理学
CiteScore
1.80
自引率
10.00%
发文量
50
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.
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