Fish Innate Immune System Group, Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100, Murcia, Spain.
Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, 60064, USA.
BMC Genomics. 2017 Dec 2;18(1):939. doi: 10.1186/s12864-017-4349-3.
Skin and its mucus are known to be the first barrier of defence against any external stressors. In fish, skin wounds frequently appear as a result of intensive culture and also some diseases have skin ulcers as external clinical signs. However, there is no information about the changes produced by the wounds in the mucosae. In the present paper, we have studied the alterations in the proteome map of skin mucus of gilthead seabream during healing of experimentally produced chronic wounds by 2-DE followed by LC-MS/MS. The corresponding gene expression changes of some identified skin proteins were also investigated through qPCR.
Our study has successfully identified 21 differentially expressed proteins involved in immunity and stress processes as well as other metabolic and structural proteins and revealed, for the first time, that all are downregulated in the skin mucus of wounded seabream specimens. At transcript level, we found that four of nine markers (ighm, gst3, actb and krt1) were downregulated after causing the wounds while the rest of them remained unaltered in the wounded fish. Finally, ELISA analysis revealed that IgM levels were significantly lower in wounded fish compared to the control fish.
Our study revealed a decreased-expression at protein and for some transcripts at mRNA levels in wounded fish, which could affect the functionality of these molecules, and therefore, delay the wound healing process and increase the susceptibility to any infection after wounds in the skin of gilthead seabream.
众所周知,皮肤及其黏液是抵御任何外部应激源的第一道防线。在鱼类中,由于密集养殖,皮肤经常受伤,而且有些疾病的外部临床症状是皮肤溃疡。然而,关于伤口对黏液产生的变化,目前还没有信息。在本文中,我们通过 2-DE 结合 LC-MS/MS 研究了在实验性慢性伤口愈合过程中,金头鲷皮肤黏液的蛋白质组图谱的变化。还通过 qPCR 研究了一些鉴定出的皮肤蛋白的相应基因表达变化。
我们的研究成功地鉴定了 21 种差异表达蛋白,这些蛋白参与免疫和应激过程以及其他代谢和结构蛋白的过程,并首次发现所有这些蛋白在受伤的金头鲷标本的皮肤黏液中均下调。在转录水平上,我们发现 9 个标志物中的 4 个(ighm、gst3、actb 和 krt1)在造成伤口后下调,而其余标志物在受伤的鱼中保持不变。最后,ELISA 分析显示,受伤鱼的 IgM 水平明显低于对照组。
我们的研究表明,受伤鱼的蛋白质表达和一些转录本的 mRNA 水平下调,这可能会影响这些分子的功能,从而延迟金头鲷皮肤伤口的愈合过程,并增加受伤后感染的易感性。