Department of Physics and Biophysics, Institute of Biology, Warsaw University of Life Sciences, 02-776 Warsaw, Poland.
Department of Physiological Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences, 02-776 Warsaw, Poland.
Int J Mol Sci. 2021 Mar 20;22(6):3177. doi: 10.3390/ijms22063177.
TRPV1, known as a capsaicin receptor, is the best-described transient receptor potential (TRP) ion channel. Recently, it was shown to be expressed by non-excitable cells such as lymphocytes. However, the data regarding the functional expression of the TRPV1 channel in the immune cells are often contradictory. In the present study, we performed a phylogenetical analysis of the canine TRP ion channels, we assessed the expression of TRPV1 in the canine peripheral blood mononuclear cells (PBMC) by qPCR and Western blot, and we determined the functionality of TRPV1 by whole-cell patch-clamp recordings and calcium assay. We found high expression of TRPV2, -M2, and -M7 in the canine PBMCs, while expression of TRPV1, -V4 and, -M5 was relatively low. We confirmed that TRPV1 is expressed on the protein level in the PBMC and it localizes in the plasma membrane. The whole-cell patch-clamp recording revealed that capsaicin application caused a significant increase in the current density. Similarly, the results from the calcium assay show a dose-dependent increase in intracellular calcium level in the presence of capsaicin that was partially abolished by capsazepine. Our study confirms the expression of TRPV1 ion channel on both mRNA and protein levels in the canine PBMC and indicates that the ion channel is functional.
瞬时受体电位香草酸受体 1(TRPV1),又称辣椒素受体,是研究最为透彻的瞬时受体电位(TRP)离子通道之一。最近的研究表明,它在淋巴细胞等非兴奋性细胞中表达。然而,关于 TRPV1 通道在免疫细胞中的功能表达的数据往往存在矛盾。在本研究中,我们对犬类 TRP 离子通道进行了系统发育分析,通过 qPCR 和 Western blot 评估了 TRPV1 在犬外周血单核细胞(PBMC)中的表达,通过全细胞膜片钳记录和钙测定来确定 TRPV1 通道的功能。我们发现 TRPV2、-M2 和 -M7 在犬 PBMC 中高表达,而 TRPV1、-V4 和 -M5 的表达相对较低。我们证实 TRPV1 在 PBMC 中表达于蛋白水平,并且位于质膜上。全细胞膜片钳记录显示,辣椒素的应用导致电流密度显著增加。同样,钙测定的结果表明,在存在辣椒素的情况下,细胞内钙水平呈剂量依赖性增加,而这种增加部分被辣椒素拮抗剂(capsazepine)所阻断。本研究证实 TRPV1 离子通道在犬 PBMC 中同时在 mRNA 和蛋白水平上表达,并表明该离子通道具有功能。