Vicente-Carrillo Alejandro, Álvarez-Rodríguez Manuel, Castaño Cristina, Toledano-Díaz Adolfo, Martínez-Nevado Eva, Rodríguez-Martínez Heriberto, Santiago-Moreno Julián
Department of Biomedical and Clinical Sciences (BKV), Linköping University, Linköping, Sweden.
Current affiliation: Department of Animal Production, Veterinary Faculty, Complutense University of Madrid, Madrid, Spain.
Vet Med Sci. 2025 Jul;11(4):e70459. doi: 10.1002/vms3.70459.
Despite the apparent progress in reproductive technologies in wild ruminant species, healthy live births have been limited. Acquiring a sound knowledge of the molecular basis of most functional aspects of spermatozoa will improve the effectiveness of reproductive techniques and optimise conservation programs for threatened species. CatSper channels, opioid receptors and CD44 are involved in sperm capacitation of humans and domestic animals, but their presence in wild ruminants is yet undisclosed. The aim of this study was to determine the presence and localisation of CatSper 1-4, μ, δ and κ-opioid receptors and CD44 in three wild ruminant species spermatozoa (aoudad [n = 5], Iberian ibex [n = 5], mouflon [n = 5]), which show different resistance to freezing-thawing processes. Western blotting (WB) and immunocytochemistry (ICC) performed with commercially available antibodies revealed that aoudad, Iberian ibex and mouflon are equipped with the aforementioned channels and receptors, sharing localisation with other domestic animals' spermatozoa but presenting species-particularities. WB revealed homogeneous results in CatSper 1, Catsper 2, Catsper 3 and CatSper 4 among the spermatozoa of the three species, unlike μ, δ and κ-opioid and CD44 receptors that showed substantial inter-species differences in the number of bands. ICC showed inter-species differences in the location of CatSper 1-4, μ, δ and κ-opioid and CD44 receptors. Data confirmed their presence and putative role on sperm function in wild ruminant species. Inter-species differences in the location of CatSper 1-4, μ, δ and κ-opioid and CD44 receptors might underlie the variable response to reproductive technologies in these species.
尽管野生反刍动物的生殖技术取得了明显进展,但健康活产数量仍然有限。深入了解精子大多数功能方面的分子基础,将提高生殖技术的有效性,并优化濒危物种的保护计划。CatSper通道、阿片受体和CD44参与人类和家畜的精子获能,但它们在野生反刍动物中的存在尚未被揭示。本研究的目的是确定三种野生反刍动物(旋角羚[n = 5]、西班牙羱羊[n = 5]、摩弗伦羊[n = 5])精子中CatSper 1 - 4、μ、δ和κ阿片受体以及CD44的存在和定位,这三种动物对冻融过程表现出不同的抗性。使用市售抗体进行的蛋白质印迹法(WB)和免疫细胞化学法(ICC)显示,旋角羚、西班牙羱羊和摩弗伦羊具备上述通道和受体,其定位与其他家畜的精子相同,但存在物种特异性。WB显示,三种动物精子中的CatSper 1、CatSper 2、CatSper 3和CatSper 4结果一致,而μ、δ和κ阿片受体以及CD44受体在条带数量上表现出显著的种间差异。ICC显示,CatSper 1 - 4、μ、δ和κ阿片受体以及CD44受体的定位存在种间差异。数据证实了它们在野生反刍动物精子功能中的存在和假定作用。CatSper 1 - 4、μ、δ和κ阿片受体以及CD44受体定位的种间差异可能是这些物种对生殖技术反应不同的原因。