Kasica Natalia, Jakubowski Piotr, Kaleczyc Jerzy
Department of Animal Anatomy, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Oczapowskiego 13 Street, Box 105J, 10-719 Olsztyn, Poland.
Department of Pharmacology and Toxicology, School of Medicine, University of Warmia and Mazury in Olsztyn, Warszawska 30 Street, 11-041 Olsztyn, Poland.
Cells. 2021 Mar 20;10(3):690. doi: 10.3390/cells10030690.
Zebrafish has emerged as a powerful model in studies dealing with pigment development and pathobiology of pigment diseases. Due to its conserved pigment pattern with established genetic background, the zebrafish is used for screening of active compounds influencing melanophore, iridophore, and xanthophore development and differentiation. In our study, zebrafish embryos and larvae were used to investigate the influence of third-generation noncompetitive P-glycoprotein inhibitor, tariquidar (TQR), on pigmentation, including phenotype effects and changes in gene expression of chosen chromatophore differentiation markers. Five-day exposure to increasing TQR concentrations (1 µM, 10 µM, and 50 µM) resulted in a dose-dependent augmentation of the area covered with melanophores but a reduction in the area covered by iridophores. The observations were performed in three distinct regions-the eye, dorsal head, and tail. Moreover, TQR enhanced melanophore renewal after depigmentation caused by 0.2 mM 1-phenyl-2-thiourea (PTU) treatment. qPCR analysis performed in 56-h post-fertilization (hpf) embryos demonstrated differential expression patterns of genes related to pigment development and differentiation. The most substantial findings include those indicating that TQR had no significant influence on leukocyte tyrosine kinase, GTP cyclohydrolase 2, tyrosinase-related protein 1, and forkhead box D3, however, markedly upregulated tyrosinase, dopachrome tautomerase and melanocyte inducing transcription factor, and downregulated purine nucleoside phosphorylase 4a. The present study suggests that TQR is an agent with multidirectional properties toward pigment cell formation and distribution in the zebrafish larvae and therefore points to the involvement of P-glycoprotein in this process.
斑马鱼已成为研究色素发育和色素疾病病理生物学的强大模型。由于其具有保守的色素模式和既定的遗传背景，斑马鱼被用于筛选影响黑素细胞、虹彩细胞和黄色素细胞发育及分化的活性化合物。在我们的研究中，斑马鱼胚胎和幼体被用于研究第三代非竞争性P-糖蛋白抑制剂他林洛尔（TQR）对色素沉着的影响，包括表型效应以及所选色素细胞分化标志物基因表达的变化。将斑马鱼暴露于浓度递增的TQR（1 µM、10 µM和50 µM）中5天，结果显示黑素细胞覆盖面积呈剂量依赖性增加，而虹彩细胞覆盖面积减少。这些观察在三个不同区域——眼睛、头部背面和尾部进行。此外，TQR增强了由0.2 mM 1-苯基-2-硫脲（PTU）处理引起的色素脱失后的黑素细胞更新。对受精后56小时（hpf）胚胎进行的qPCR分析表明，与色素发育和分化相关的基因存在差异表达模式。最显著的发现包括，TQR对白细胞酪氨酸激酶、GTP环化水解酶2、酪氨酸酶相关蛋白1和叉头框D3没有显著影响，然而，它显著上调了酪氨酸酶、多巴色素互变酶和黑素细胞诱导转录因子，并下调了嘌呤核苷磷酸化酶4a。本研究表明，TQR是一种对斑马鱼幼体色素细胞形成和分布具有多向性作用的物质，因此表明P-糖蛋白参与了这一过程。
