鸡弱精症相关精浆蛋白组分析

Seminal Plasma Proteomics Analysis Associated with Asthenospermia in Rooster

旨在对鸡精浆蛋白组进行分析,筛选与弱精症相关的蛋白。本试验选择同批次孵化的宁都黄鸡公鸡进行精液品质检测,每隔3 d进行1次镜检,共进行3次。选择前向精子比例<20%,且直线前向精子比例<10%的个体作为试验样本,前向精子比例>80%,且直线前向精子比例>40%的个体为对照样本,每组各4只。利用液相色谱-质谱(LC-MS/MS)技术开展鸡精浆蛋白组学的研究,筛选与弱精症相关的差异蛋白。结果显示,宁都黄鸡弱精组的精子活力和密度极显著低于对照组(P<0.01)。两组间共筛选到差异表达蛋白78个,5个差异蛋白在弱精组中表现为上调,其余差异蛋白表现为下调。角蛋白9(KRT 9)和硒蛋白P-2(SEPP 2)仅在弱精组中检测到表达;4-羟基-2-氧戊二酸醛缩酶(HOGA 1)、苯丙氨酸羟化酶(PAH)和卵磷脂胆固醇酰基转移酶(LCAT)仅在对照组中检测到表达。经GO注释和KEGG通路分析,差异蛋白功能集中于新陈代谢、糖酵解、碳代谢和氨基酸合成等生物途径。本研究筛选的宁都黄鸡弱精症和正常公鸡精浆差异表达蛋白可作为今后鸡弱精症筛查和治疗的候选靶点,通过进一步功能分析与验证为阐明弱精症的发病机理提供依据。

The study aimed to analyze the proteomics of chicken seminal plasma and screen the differential proteins related to asthenospermia.The experimental roosters were selected from Ningdu yellow chicken hatched in the same batch,and semen quality was examined every 3 days,3 times in total.Four individuals with forward sperm ratio<20%and straight forward sperm ratio<10%were selected as asthenospermia group.Four individuals with forward sperm ratio>80%and straight forward sperm ratio>40%were selected as control group.Using liquid chromatography-mass spectrometry(LC-MS/MS)technology,the proteomics of chicken seminal plasma was studied and differential proteins related to asthenospermia were preliminary screened.The results showed that the sperm motility and density of roosters in asthenospermia group were extremely significantly lower than those in control group(P<0.01).A total of 78 differentially expressed proteins were screened out between the two groups.Compared with control group,5 differential proteins were up-regulated and the others were down-regulated in the asthenospermia group.It is important to note that the expression of keratin 9(KRT 9)and selenoprotein P-2(SEPP 2)were only detected in asthenospermia group,and the expression of 4-hydroxy-2-oxoglutaracturonase(HOGA 1),phenylalanine hydroxylase(PAH)and lecithin cholesterol acyltransferase(LCAT)were only detected in the control group.According to GO and KEGG analysis,the functions of differential proteins were mainly focused on metabolism,glycolysis,carbon metabolism,amino acid synthesis pathways,and so on.The differentially expressed proteins in seminal plasma of Ningdu yellow chickens in asthenospermia and control groups screened in this study could be used as candidate targets for screening and treatment of asthenospermia in chickens.The further functional analysis and verification will provide a basis for elucidating the pathogenesis of asthenospermia.