Atikuzzaman Mohammad, Sanz Libia, Pla Davinia, Alvarez-Rodriguez Manuel, Rubér Marie, Wright Dominic, Calvete Juan J, Rodriguez-Martinez Heriberto
Department of Clinical and Experimental Medicine, University of Linköping, Linköping, Sweden.
Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain.
Comp Biochem Physiol Part D Genomics Proteomics. 2017 Mar;21:27-40. doi: 10.1016/j.cbd.2016.10.006. Epub 2016 Nov 3.
The high egg-laying capacity of the modern domestic chicken (i.e. White Leghorn, WL) has arisen from the low egg-laying ancestor Red Junglefowl (RJF) via continuous trait selection and breeding. To investigate whether this long-term selection impacted the seminal fluid (SF)-proteome, 2DE electrophoresis-based proteomic analyses and immunoassays were conducted to map SF-proteins/cytokines in RJF, WL and a 9th generation Advanced Intercross Line (AIL) of RJF/WL-L13, including individual SF (n=4, from each RJF, WL and AIL groups) and pools of the SF from 15 males of each group, analyzed by 2DE to determine their degree of intra-group (AIL, WL, and RJF) variability using Principal Component Analysis (PCA); respectively an inter-breed comparative analysis of intergroup fold change of specific SF protein spots intensity between breeds. The PCA clearly highlighted a clear intra-group similarity among individual roosters as well as a clear inter-group variability (e.g. between RJF, WL and AIL) validating the use of pools to minimize confounding individual variation. Protein expression varied considerably for processes related to sperm motility, nutrition, transport and survival in the female, including signaling towards immunomodulation. The major conserved SF-proteins were serum albumin and ovotransferrin. Aspartate aminotransferase, annexin A5, arginosuccinate synthase, glutathione S-transferase 2 and l-lactate dehydrogenase-A were RJF-specific. Glyceraldehyde-3-phosphate dehydrogenase appeared specific to the WL-SF while angiotensin-converting enzyme, γ-enolase, coagulation factor IX, fibrinogen α-chain, hemoglobin subunit α-D, lysozyme C, phosphoglycerate kinase, Src-substrate protein p85, tubulins and thioredoxin were AIL-specific. The RJF-SF contained fewer immune system process proteins and lower amounts of the anti-inflammatory/immunomodulatory TGF-β2 compared to WL and AIL, which had low levels- or lacked pro-inflammatory CXCL10 compared to RJF. The seminal fluid proteome differs between ancestor and modern chicken, with a clear enrichment of proteins and peptides related to immune-modulation for sperm survival in the female and fertility.
现代家鸡（即白来航鸡，WL）的高产蛋能力是通过持续的性状选择和育种，从低产蛋的祖先原鸡（RJF）进化而来的。为了研究这种长期选择是否影响了精液（SF）蛋白质组，我们进行了基于二维电泳的蛋白质组分析和免疫测定，以绘制RJF、WL以及RJF/WL-L13的第9代高级杂交系（AIL）中的SF蛋白质/细胞因子图谱，包括个体精液（每组n = 4，分别来自RJF、WL和AIL组）以及每组15只雄性的精液池，通过二维电泳进行分析，使用主成分分析（PCA）确定它们在组内（AIL、WL和RJF）的变异程度；分别对各品种间特定SF蛋白斑点强度的组间倍数变化进行品种间比较分析。PCA清楚地突出了个体公鸡之间明显的组内相似性以及明显的组间变异性（例如RJF、WL和AIL之间），验证了使用精液池以最小化个体差异的干扰。与精子活力、营养、运输以及在雌性体内的存活相关的过程中，蛋白质表达差异很大，包括免疫调节信号传导。主要的保守SF蛋白是血清白蛋白和卵转铁蛋白。天冬氨酸转氨酶、膜联蛋白A5、精氨琥珀酸合酶、谷胱甘肽S-转移酶2和L-乳酸脱氢酶-A是RJF特有的。甘油醛-3-磷酸脱氢酶似乎是WL精液特有的，而血管紧张素转换酶、γ-烯醇化酶、凝血因子IX、纤维蛋白原α链、血红蛋白亚基α-D、溶菌酶C、磷酸甘油酸激酶、Src底物蛋白p85、微管蛋白和硫氧还蛋白是AIL特有的。与WL和AIL相比，RJF精液中免疫系统相关蛋白较少，抗炎/免疫调节性TGF-β2含量较低，而WL和AIL与RJF相比，促炎CXCL10水平较低或缺乏。祖先鸡和现代鸡的精液蛋白质组不同，与免疫调节相关的蛋白质和肽明显富集，以促进精子在雌性体内的存活和生育能力。
