Laboratório de Biologia Celular e do Desenvolvimento (LABID), Departamento de Biologia Molecular, Universidade Federal da Paraíba, João Pessoa, Paraíba, Brazil.
Laboratoire des Sciences de l'Environnement Marin (LEMAR), UMR 6539 CNRS UBO IRD IFREMER, Institut Universitaire Européen de la Mer, Technopôle Brest-Iroise, Rue Dumont d'Urville, 29280 Plouzané, France.
Fish Shellfish Immunol. 2017 Nov;70:195-203. doi: 10.1016/j.fsi.2017.09.014. Epub 2017 Sep 5.
ABC transporters activity and expression have been associated with the multixenobiotic resistance phenotype (MXR). The activity of these proteins leads to a reduction in the intracellular concentration of several xenobiotics, thus reducing their toxicity. However, little attention has been given to the expression of ABC transporters in marine invertebrates and few studies have investigated their role in immune system cells of sea urchins and shellfish bivalves. The aim of the present study was to investigate the activity of the ABC transporters ABCB1 and ABCC1 in immune system cells of sea urchins (coelomocytes) and oysters (hemocytes) from different climatic regions (Brazil and France). Sea urchins and oysters were collected at Paraíba coast; Brazil (Echinometra lucunter and Crassostrea gasar) and Rade of Brest; France (Echinus esculentus and Crassostrea gigas). Coelomocytes and hemocytes were stained with the ABC transporter substrate calcein-AM and dye accumulation analyzed under flow cytometry. Reversin 205 (ABCB1 transporter blocker) and MK571 (ABCC1 transporter blocker) were used as pharmacological tools to investigate ABC transporter activity. A different pattern of calcein accumulation was observed in coelomocytes: phagocytes > colorless spherulocytes > vibrate cells > red spherulocytes. The treatment with MK571 increased calcein fluorescence levels in coelomocytes from both species. However, reversin 205 treatment was not able to increase calcein fluorescence in E. esculentus coelomocytes. These data suggest that ABCC1-like transporter activity is present in both sea urchin species, but ABCB1-like transporter activity might only be present in E. lucunter coelomocytes. The activity of ABCC1-like transporter was observed in all cell types from both bivalve species. However, reversin 205 only increased calcein accumulation in hyalinocytes of the oyster C. gasar, suggesting the absence of ABCB1-like transporter activity in all other cell types, including hyalinocytes from the oyster C. gigas. Additionally, our results showed that C. gigas exhibited higher activity of ABCC1-like transporter in all hemocyte types than C. gasar. The present work is the first to characterize ABCB1 and ABCC1-like transporter activity in the immune system cells of sea urchins E. lucunter and E. esculentus and oysters. Our findings encourage the performing studies regarding ABC transporters activity/expression in immune system cells form marine invertebrates under stress conditions and the possible use of ABC transporters as biomarkers.
ABC 转运蛋白的活性和表达与多药耐药表型(MXR)有关。这些蛋白质的活性导致几种外源化学物质的细胞内浓度降低,从而降低其毒性。然而,人们对海洋无脊椎动物中 ABC 转运蛋白的表达关注甚少,很少有研究调查它们在海胆和贝类双壳类动物的免疫系统细胞中的作用。本研究旨在研究来自不同气候区(巴西和法国)的海胆(体腔细胞)和牡蛎(血细胞)的 ABC 转运蛋白 ABCB1 和 ABCC1 的活性。在巴西 Paraíba 海岸(Echinometra lucunter 和 Crassostrea gasar)和法国 Rade of Brest 收集了海胆和牡蛎。用 ABC 转运体底物 calcein-AM 染色体腔细胞和血细胞,并通过流式细胞术分析染料积累。使用 Reversin 205(ABCB1 转运体阻断剂)和 MK571(ABCC1 转运体阻断剂)作为药理学工具来研究 ABC 转运体的活性。在体腔细胞中观察到不同的 calcein 积累模式:吞噬细胞>无色球形细胞>振动细胞>红色球形细胞。MK571 处理增加了两种物种的体腔细胞中的 calcein 荧光水平。然而,reversin 205 处理不能增加 E. esculentus 体腔细胞中的 calcein 荧光。这些数据表明,ABCC1 样转运体活性存在于两种海胆物种中,但 ABCB1 样转运体活性可能仅存在于 E. lucunter 体腔细胞中。在两种贝类的所有细胞类型中均观察到 ABCC1 样转运体的活性。然而,reversin 205 仅增加了牡蛎 C. gasar 透明细胞中的 calcein 积累,表明 ABCB1 样转运体活性不存在于所有其他细胞类型中,包括牡蛎 C. gigas 的透明细胞。此外,我们的结果表明,C. gigas 在所有血细胞类型中的 ABCC1 样转运体活性均高于 C. gasar。本研究首次描述了海胆 E. lucunter 和 E. esculentus 以及牡蛎的免疫系统细胞中 ABCB1 和 ABCC1 样转运体的活性。我们的发现鼓励在应激条件下对海洋无脊椎动物的免疫系统细胞中的 ABC 转运体活性/表达进行研究,并可能将 ABC 转运体用作生物标志物。