Dratwa-Chałupnik Alicja, Wojdyła Katarzyna, Ożgo Małgorzata, Lepczyński Adam, Michałek Katarzyna, Herosimczyk Agnieszka, Rogowska-Wrzesińska Adelina
Department of Physiology, Cytobiology and Proteomics, Faculty of Biotechnology and Animal Husbandry, West Pomeranian University of Technology Szczecin, Klemensa Janickiego 29, 71-270 Szczecin, Poland.
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, M-DK-5230 Odense, Denmark.
Animals (Basel). 2020 Jul 24;10(8):1257. doi: 10.3390/ani10081257.
Urine is a biological diagnostic material suitable not only for the analysis of kidney and urinary tract functions but also the function of other tissues and organs. The urine proteome of adult mammals differs from the urine proteome of neonatal ones. The establishment of urinary protein maps of healthy newborn calves is important for diagnosing and monitoring the progression of various diseases. The experiment was carried out on a Polish-Friesian var. of Black-and-White male calves in the sixth day of postnatal life. The two proteomics approaches used for separation and identification of urinary proteins were: 2-DE with MALDI-TOF-TOF-MS/MS and 1-DE with LC-MS/MS. This resulted in the identification of 692 urinary proteins. The majority of them were classified as extracellular proteins (40.32%), as well as proteins involved in regulation of major cellular processes (31.07%). We have observed the presence of unique proteins associated with embryonic (ameloblastin, alpha-fetoprotein, Delta-like protein, embryo-specific fibronectin 1 transcript variant, Indian hedgehog homolog) and kidney development (angiotensin-converting enzyme, angiotensinogen, aquaporin-1, calbindin, glypican 3, nidogen 1, pro-cathepsin H). Additionally, proteins involved in the renal regulation of water and electrolyte balance (angiotensinogen, angiotensin-converting enzyme, aquaporin-1, ezrin, uromodulin) were detected. Presented in the current study 1-D and 2-D urinary proteomic maps are the basis for the identification and detection of prognostic biomarkers important for defining a calf's health status.
尿液是一种生物诊断材料,不仅适用于分析肾脏和泌尿系统的功能,还适用于分析其他组织和器官的功能。成年哺乳动物的尿液蛋白质组与新生哺乳动物的尿液蛋白质组不同。建立健康新生犊牛的尿蛋白图谱对于诊断和监测各种疾病的进展具有重要意义。该实验在出生后第六天的波兰黑白花雄性犊牛品种上进行。用于分离和鉴定尿蛋白的两种蛋白质组学方法是:二维凝胶电泳结合基质辅助激光解吸电离飞行时间串联质谱(2-DE 与 MALDI-TOF-TOF-MS/MS)和一维凝胶电泳结合液相色谱-质谱联用(1-DE 与 LC-MS/MS)。这导致鉴定出692种尿蛋白。其中大多数被归类为细胞外蛋白(40.32%),以及参与主要细胞过程调节的蛋白(31.07%)。我们观察到存在与胚胎(成釉蛋白、甲胎蛋白、Delta样蛋白、胚胎特异性纤连蛋白1转录变体、印度刺猬同源物)和肾脏发育(血管紧张素转换酶、血管紧张素原、水通道蛋白-1、钙结合蛋白、磷脂酰肌醇蛋白聚糖3、巢蛋白1、组织蛋白酶H前体)相关的独特蛋白质。此外,还检测到参与肾脏水和电解质平衡调节的蛋白质(血管紧张素原、血管紧张素转换酶、水通道蛋白-1、埃兹蛋白、尿调节蛋白)。本研究中呈现的一维和二维尿蛋白质组图谱是鉴定和检测对确定犊牛健康状况重要的预后生物标志物的基础。