3C). The data also suggest that cationic dyes (YO-PRO-1 and PI) do not Selleckchem Ponatinib pass through plasma

membranes even after ATP stimulation. Given the presence of separate P2X7R-mediated permeation pathways for cationic and anionic dyes [18], it still remains unclear whether the anionic dyes are taken up on ATP stimulation in LPS-primed swine kidney macrophages. We further tested the effect of extracellular ATP in swine liver-derived macrophages. ATP-induced P2X7R-mediated sustained Ca2+ influx (Fig. 4A), and the maturation and release of IL-1β (B) were also not observed in swine liver macrophages despite the presence of P2X7R mRNA (C) and protein (D). ATP-induced P2X7R-mediated cellular events, such as sustained Ca2+ influx, the maturation and release of IL-1β, and membrane pore formation, were not observed in swine kidney and liver-derived macrophages stimulated with ATP despite the fact that they express P2X7R mRNA and protein. The data raise the possibility that P2X7R is non-functional in these swine macrophages at least under the culture conditions used in the present study, although amino acid sequence of swine P2X7R is 88% and 79% similar

to the sequences of human and mouse P2X7R, respectively. The precise LEE011 molecular weight mechanisms explaining the non-responsiveness of P2X7R to ATP in swine macrophages remain to be elucidated. Since the functional expression of P2X7R has been reported in swine ovary theca cells [9], this may be due to differences in tissue and/or cell types. It is also likely that the presence of alternative splice variants of P2X7R may be related to the non-functional expression of P2X7R in swine macrophages. In fact, several P2X7R splice variants have been defined in human and rodents [19] and [20]. Although we have confirmed the expression of full length of swine P2X7R by immunoblotting (around 75-kDa band in Fig. 1 and Fig. 4) and DNA sequence analysis of 1785 bp amplified product obtained by RT-PCR (data

not shown), future experiments will be required to determine whether other P2X7R splice variants are expressed in 2-hydroxyphytanoyl-CoA lyase swine macrophages. Given the importance of P2X7R in innate immune defense in various animal species, our findings might provide new insights into the regulation of the innate immune system in swine. The authors would like to thank the staff of the Swine Management Section of the National Institute of Livestock and Grassland Science for taking care of the pigs. This study was supported by a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan (Genomic-based Technology for Agricultural Improvement, AGB-1004) and a Grant-in-Aid for Scientific Research (Category C: Grant# 25450521) from the Japan Society for the Promotion of Science (JSPS). “
“Liver-macrophages known as Kupffer cells are the resident macrophages in the liver [1].