Table 2_Broiler breeder putative lipid biomarkers associated with sperm mobility.xlsx

IntroductionBiomarkers indicative of sperm mobility in broiler breeders would provide the ability to screen for fertility potential, with a positive correlation established between sperm mobility and fertilization potential. This study characterized the lipidome of seminal plasma (SP), sperm cell (SC), and whole semen (WS) isolated from broiler breeder roosters with different sperm mobility phenotypes across key timepoints of the semen production cycle.
MethodsWS samples were collected from five high mobility roosters and five low mobility roosters during early, mid, and late semen production, with SP separated from SC by centrifugation. Using multiple reaction monitoring (MRM) profiling, a total of 3241 lipid species were identified in rooster semen across ten lipid classes. Metaboanalyst 6.0 was used to analyze the relative ion intensity for each lipid species due to sperm mobility phenotype through a t-test and due to timepoint through a one-way ANOVA, with lipid ontology enrichment analysis performed using LION. Metaboanalyst 6.0 was also used to perform biomarker analysis for the sperm mobility phenotype in WS samples.
ResultsLipid class total abundance differed with sample type, sperm mobility phenotype, and timepoint. A total of 31, 99, and 112 lipid species were found to be different between low and high mobility males across timepoints in the SP, SC, and WS samples, respectively. Lipid ontology enrichment analysis revealed stark contrasts in lipid-based functions key to sperm survival, storage, and productivity between low and high sperm mobility phenotypes. Through biomarker analysis, 8 lipid species were identified as excellent sperm mobility biomarkers that could be detected in early and mid-semen production.
DiscussionTimepoint based changes in lipid species were unique to each sperm mobility phenotype, with low sperm mobility roosters exhibiting a larger number of lipid species changes over the semen production cycle in the SP and SC when compared to high sperm mobility roosters. This is the first study to characterize poultry semen lipidome using MRM profiling. The lipid species identified between low and high sperm mobility roosters could be utilized in the poultry industry as potential biomarkers of fertility potential, with the ability to screen for the economical trait of fertility potential early in semen production.

引言：可指示肉鸡种公鸡精子活力的生物标志物，将助力筛选受精潜力相关性状，目前已有研究证实精子活力与受精潜能呈正相关。本研究针对精液生产周期关键时间点、具有不同精子活力表型的肉鸡种公鸡，对其精浆（seminal plasma, SP）、精子细胞（sperm cell, SC）及全精液（whole semen, WS）的脂质组（lipidome）进行了表征分析。
方法：本研究于精液生产的早期、中期与晚期，分别从5只高活力公鸡与5只低活力公鸡中采集全精液（whole semen, WS）样本，并通过离心法分离精浆与精子细胞。采用多反应监测（multiple reaction monitoring, MRM）谱分析技术，本研究在公鸡精液的10个脂质类别中共鉴定出3241种脂质分子。使用Metaboanalyst 6.0软件，针对不同精子活力表型下各脂质分子的相对离子强度开展t检验，针对不同采集时间点开展单因素方差分析（one-way ANOVA）；同时借助LION工具完成脂质本体富集分析。此外，本研究还通过Metaboanalyst 6.0对WS样本中的精子活力表型进行生物标志物筛选分析。
结果：脂质类别的总丰度会随样本类型、精子活力表型及采集时间点发生变化。在不同时间点的精浆、精子细胞及全精液样本中，分别鉴定出31、99和112种在高低活力公鸡间存在显著差异的脂质分子。脂质本体富集分析结果显示，在精子存活、储存及生产性能相关的脂质功能层面，高低精子活力表型组间存在显著差异。通过生物标志物筛选分析，共鉴定出8种优质精子活力生物标志物，可在精液生产早期与中期被检测到。
讨论：脂质分子随采集时间点的变化模式因精子活力表型而异；与高活力公鸡相比，低活力公鸡的精浆与精子细胞中，在精液生产周期内出现丰度变化的脂质分子数量更多。本研究是首次采用MRM谱分析技术表征家禽精液脂质组的相关研究。本研究鉴定出的高低活力公鸡间差异脂质分子，可在家禽养殖业中作为受精潜力的潜在生物标志物，实现在精液生产早期就对该经济性状进行筛选。