Mendoza Noelia, Casao Adriana, Domingo Juan, Quintín Francisco, Laviña Adolfo, Fantova Enrique, Cebrián-Pérez José Álvaro, Muiño-Blanco Teresa, Pérez-Pe Rosaura
Grupo BIOFITER-Departamento de Bioquímica y Biología Molecular y Celular-Instituto Universitario de Investigación en Ciencias Ambientales de Aragón (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain.
Centro de Transferencia Agroalimentaria (Gobierno de Aragón), Zaragoza, Spain.
Front Vet Sci. 2021 May 7;8:650572. doi: 10.3389/fvets.2021.650572. eCollection 2021.
The prediction of the fertilizing ability of a seminal dose continues to be a primary aim in the field of artificial insemination (AI). To achieve this goal, in this study we have included the evaluation of some non-conventional sperm quality markers. A total of 3,906 ewes from 52 different farms were inseminated with 357 refrigerated seminal doses obtained from 45 mature rams. The same samples were used for sperm quality analysis including membrane integrity, capacitation status, oxygen consumption and apoptotic-like markers such as phosphatidylserine translocation (PS), plasmalemma disorganization/mitochondrial membrane potential, caspase activation and DNA damage. Seminal doses from the breeding (B) season presented higher percentages of intact membrane (IM), non permeant (NP) membrane with high mitochondrial membrane potential (ΔΨm) and IM without PS translocation spermatozoa than those from the non-breeding (NB) season. Therefore, we can conclude that there were less spermatozoa showing apoptotic-like features in the seminal doses from the B than the NB season, although these differences did not affect field fertility. Only the percentage of intact membrane, non-capacitated (IM-NC) spermatozoa showed a significant correlation with fertility ( = 0.005) and fecundity ( = 0.007) values obtained after cervical AI when all data were evaluated. When the data were sorted by season and distance to the farms where AI was performed, the correlation between the percentage of IM-NC spermatozoa and reproductive parameters increased in the NB season and progressively with remoteness from the farms. Some other sperm parameters, like NP with high ΔΨm, IM sperm without active caspases and DNA-intact spermatozoa, also showed significant correlations with the reproductive parameters in the sorted data. Moreover, the increment in both the percentage of IM-NC and DNA-intact spermatozoa would increase the probability of obtaining a fertility higher than the mean (>52%), as revealed by a multiple logistic regression analysis. In conclusion, we have identified two seminal markers-the percentage of intact membrane, non-capacitated spermatozoa, and DNA intact spermatozoa-which could be used as a test to discard males in AI programs, which is highly important from an economic point of view and can contribute to achieving satisfactory fertility rates.
精液剂量受精能力的预测仍然是人工授精(AI)领域的主要目标。为实现这一目标,在本研究中,我们纳入了一些非常规精子质量标志物的评估。来自52个不同农场的3906只母羊用从45只成年公羊获得的357份冷冻精液剂量进行了授精。相同的样本用于精子质量分析,包括膜完整性、获能状态、氧消耗以及凋亡样标志物,如磷脂酰丝氨酸易位(PS)、质膜解体/线粒体膜电位、半胱天冬酶激活和DNA损伤。繁殖(B)季节的精液剂量中,完整膜(IM)、具有高线粒体膜电位(ΔΨm)的非通透(NP)膜以及无PS易位精子的IM精子百分比高于非繁殖(NB)季节。因此,我们可以得出结论,B季节精液剂量中显示凋亡样特征的精子比NB季节少,尽管这些差异并未影响田间受精率。当评估所有数据时,仅完整膜、未获能(IM-NC)精子的百分比与经宫颈人工授精后获得的受精率(=0.005)和繁殖力(=0.007)值呈显著相关。当按季节和与进行人工授精的农场的距离对数据进行分类时,IM-NC精子百分比与生殖参数之间的相关性在NB季节增加,并随着与农场距离的增加而逐渐增加。在分类数据中,其他一些精子参数,如具有高ΔΨm的NP、无活性半胱天冬酶的IM精子和DNA完整的精子,也与生殖参数呈显著相关。此外,多元逻辑回归分析表明,IM-NC和DNA完整精子百分比的增加都会增加获得高于平均受精率(>52%)的概率。总之,我们确定了两种精液标志物——完整膜、未获能精子的百分比和DNA完整的精子——可用于在人工授精计划中筛选雄性,这从经济角度来看非常重要,并且有助于实现令人满意的受精率。