Effect of exogenous lactoferrin and phosphoglycerate mutase 2 on the cryopreservation of wild ruminant epididymal/ejaculated sperm and testicular tissue

IF 2.3 3区生物学 Q2 BIOLOGY

Cryobiology Pub Date : 2024-12-01 DOI:10.1016/j.cryobiol.2024.105182

Julián Santiago-Moreno , Patricia Peris-Frau , Adolfo Toledano-Díaz , Cristina Castaño , Rosario Velázquez , Esther Alba , Félix Gómez-Guillamón , Leonor Camacho , Paloma Prieto , Belén Martínez-Madrid

{"title":"Effect of exogenous lactoferrin and phosphoglycerate mutase 2 on the cryopreservation of wild ruminant epididymal/ejaculated sperm and testicular tissue","authors":"Julián Santiago-Moreno , Patricia Peris-Frau , Adolfo Toledano-Díaz , Cristina Castaño , Rosario Velázquez , Esther Alba , Félix Gómez-Guillamón , Leonor Camacho , Paloma Prieto , Belén Martínez-Madrid","doi":"10.1016/j.cryobiol.2024.105182","DOIUrl":null,"url":null,"abstract":"<div><div>Spermatozoa collected from the cauda epididymis of wild ruminants are more cryoresistant than are ejaculated spermatozoa. This work examines the effects of lactoferrin (LF) and phosphoglycerate mutase 2 (PGAM2), which are abundant in the epididymal sperm of wild ruminants, as additives in Iberian ibex and mouflon sperm extenders. In addition, LF was added to a vitrification medium to determine whether it also provided protection during the cryopreservation of testicular tissue. Epididymal sperm samples and testes were recovered post-mortem from ibexes (n = 13) and mouflons (n = 8) during the breeding season. Ejaculates were collected from both species (n = 10 each) using the transrectal ultrasound-guided massage technique. Four aliquots were taken from each sample, diluted in the appropriate freezing extender for each species, and subjected to the following treatments: control, LF (100 μg/ml), PGAM2 (25 μg/ml), and LF + PGAM2. Testicular tissue was cut into small pieces and cryopreserved by needle-immersed vitrification in a medium with or without LF. <em>In vivo</em> fertilization capacity was assessed using frozen-thawed ejaculated sperm from the ibexes. Supplementation of extenders with LF or PGAM2, and their combination, had no beneficial or harmful effect on any sperm variable after freezing-thawing. Artificial insemination of ibexes showed that the fertility rate in controls was 62.5 %, but this fell to 20 % in females inseminated with sperm treated with LF (p = 0.06), suggesting a putative negative effect of LF on fertility. The viability of elongated spermatids and spermatozoa from mouflon testes was greater (p < 0.05) in samples that were vitrified-warmed with LF. Thus, the hypothesis that supplementing ejaculated sperm with LF and/or PGAM2 improves sperm quality after freeze-thawing was not upheld. However, LF would seem an appropriate additive when vitrifying mouflon testicular tissue.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105182"},"PeriodicalIF":2.3000,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cryobiology","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0011224024003377","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Spermatozoa collected from the cauda epididymis of wild ruminants are more cryoresistant than are ejaculated spermatozoa. This work examines the effects of lactoferrin (LF) and phosphoglycerate mutase 2 (PGAM2), which are abundant in the epididymal sperm of wild ruminants, as additives in Iberian ibex and mouflon sperm extenders. In addition, LF was added to a vitrification medium to determine whether it also provided protection during the cryopreservation of testicular tissue. Epididymal sperm samples and testes were recovered post-mortem from ibexes (n = 13) and mouflons (n = 8) during the breeding season. Ejaculates were collected from both species (n = 10 each) using the transrectal ultrasound-guided massage technique. Four aliquots were taken from each sample, diluted in the appropriate freezing extender for each species, and subjected to the following treatments: control, LF (100 μg/ml), PGAM2 (25 μg/ml), and LF + PGAM2. Testicular tissue was cut into small pieces and cryopreserved by needle-immersed vitrification in a medium with or without LF. In vivo fertilization capacity was assessed using frozen-thawed ejaculated sperm from the ibexes. Supplementation of extenders with LF or PGAM2, and their combination, had no beneficial or harmful effect on any sperm variable after freezing-thawing. Artificial insemination of ibexes showed that the fertility rate in controls was 62.5 %, but this fell to 20 % in females inseminated with sperm treated with LF (p = 0.06), suggesting a putative negative effect of LF on fertility. The viability of elongated spermatids and spermatozoa from mouflon testes was greater (p < 0.05) in samples that were vitrified-warmed with LF. Thus, the hypothesis that supplementing ejaculated sperm with LF and/or PGAM2 improves sperm quality after freeze-thawing was not upheld. However, LF would seem an appropriate additive when vitrifying mouflon testicular tissue.

查看原文本刊更多论文

外源性乳铁蛋白和磷酸甘油酸突变酶2对野生反刍动物附睾/射精精子和睾丸组织冷冻保存的影响。

从野生反刍动物附睾尾部采集的精子比射精精子具有更强的抗冻性。本研究考察了野生反刍动物附睾精子中丰富的乳铁蛋白（LF）和磷酸甘油酸突变酶2 （PGAM2）作为伊比利亚野山羊和摩富龙精子扩展剂添加剂的作用。此外，将LF添加到玻璃化培养基中，以确定其在睾丸组织冷冻保存期间是否也提供保护。在繁殖季节，从野山羊（n=13）和麋鹿（n=8）的死后提取附睾精子和睾丸样本。采用经直肠超声引导按摩技术收集两种动物的射精（各10只）。每个样品取四等分，用相应的冷冻扩展剂稀释，分别进行对照、LF （100 μg/ml）、PGAM2 （25 μg/ml）和LF+PGAM2处理。将睾丸组织切成小块，在含或不含LF的培养基中通过针浸玻璃化冷冻保存。体内受精能力评估使用冻融射精的精子从ibexx。添加LF或PGAM2及其组合对冻融后精子的任何变量均无有益或有害影响。人工授精的结果表明，对照ibex的受精率为62.5%，而经LF处理的精子受精率为20% (p = 0.06)，表明LF可能对其受精率有负面影响。用LF玻璃化加热的样品中，长形精子和摩弗龙睾丸精子的存活率更高（p < 0.05）。因此，用LF和/或PGAM2补充射精精子可以提高冷冻解冻后精子质量的假设不成立。然而，在玻璃化摩弗龙睾丸组织时，LF似乎是一种合适的添加剂。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Cryobiology 生物-生理学

CiteScore

5.40

自引率

7.40%

发文量

审稿时长

56 days

期刊介绍： Cryobiology: International Journal of Low Temperature Biology and Medicine publishes research articles on all aspects of low temperature biology and medicine. Research Areas include: • Cryoprotective additives and their pharmacological actions • Cryosurgery • Freeze-drying • Freezing • Frost hardiness in plants • Hibernation • Hypothermia • Medical applications of reduced temperature • Perfusion of organs • All pertinent methodologies Cryobiology is the official journal of the Society for Cryobiology.