PDF(Pubmed)
Abstract：
OBJECTIVE: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers.
METHODS: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate.
RESULTS: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality.
CONCLUSIONS: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.
METHODS: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate.
RESULTS: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality.
CONCLUSIONS: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.
摘要：
■本研究旨在鉴定马杜拉公牛精子中的HSP70-2和PRM1mRNA和蛋白，并证明它们作为公牛生育力生物标志物的关系。
■根据首次服务受胎率（％FSCR）的百分比将马都拉公牛的生育率分组为高生育率（HF）（79.04％；n=4），和低生育率（LF）（65.84％；n=4）。通过RT-qPCR测定了以肽基丙氨酰基异构酶A（PPIA）为管家基因的HSP70-2和PRM1的mRNA，而ELISA用于测量蛋白质丰度。在解冻后的精液样本中，精子运动性,生存能力，顶体完整性,并对精子DNA碎片指数进行分析。对精液质量的测量参数进行数据分析，相对mRNA表达，在单向方差分析中，在具有不同生育力水平（HF和LF）的公牛中，HSP70-2和PRM1的蛋白质丰度。采用Pearson相关性分析精液质量与精液质量、mRNA蛋白质，和生育率。
■检测HSP70-2和PRM1的相对mRNA表达和蛋白丰度，发现在高生育力的公牛中高表达（p<0.05），并且与精液质量的几个参数有关。
■HSP70-2和PRM1mRNA和蛋白质分子具有很大的潜力，可作为确定公牛生育力的分子标记。
■根据首次服务受胎率（％FSCR）的百分比将马都拉公牛的生育率分组为高生育率（HF）（79.04％；n=4），和低生育率（LF）（65.84％；n=4）。通过RT-qPCR测定了以肽基丙氨酰基异构酶A（PPIA）为管家基因的HSP70-2和PRM1的mRNA，而ELISA用于测量蛋白质丰度。在解冻后的精液样本中，精子运动性,生存能力，顶体完整性,并对精子DNA碎片指数进行分析。对精液质量的测量参数进行数据分析，相对mRNA表达，在单向方差分析中，在具有不同生育力水平（HF和LF）的公牛中，HSP70-2和PRM1的蛋白质丰度。采用Pearson相关性分析精液质量与精液质量、mRNA蛋白质，和生育率。
■检测HSP70-2和PRM1的相对mRNA表达和蛋白丰度，发现在高生育力的公牛中高表达（p<0.05），并且与精液质量的几个参数有关。
■HSP70-2和PRM1mRNA和蛋白质分子具有很大的潜力，可作为确定公牛生育力的分子标记。
