Blomqvist Maria, Osterbye Thomas, Månsson Jan-Eric, Horn Thomas, Buschard Karsten, Fredman Pam
Institute of Clinical Neuroscience, Experimental Neuroscience Section, The Sahlgrenska Academy at Göteborg University, Sahlgrenska University Hospital/Mölndal, SE-431 80 Mölndal, Sweden.
Glycoconj J. 2002 Jul;19(6):403-13. doi: 10.1023/B:GLYC.0000004012.14438.e6.
Previous studies using pancreas from various mammals and freshly isolated islets from rat pancreas have provided evidence supporting possible involvement of the glycosphingolipid sulfatide in insulin processing and secretion. In this study, sulfatide expression and metabolism in the beta cell line RINr1046-38 (RIN-38), commonly used as a model for beta cell functional studies, were investigated and compared with previous findings from freshly isolated islets. RIN-38 cells expressed similar amounts (2.7 +/- 1.1 nmol/mg protein, n = 19) of sulfatide as isolated rat islets and also followed the same metabolic pathway, mainly through recycling. Moreover, in agreement with findings in isolated islets, the major species of sulfatide isolated from RIN-38 cells contained C16:0 and C24:0 fatty acids. By applying subcellular isolations and electron microscopy and immunocytochemistry techniques, sulfatide was shown to be located to the secretory granules, the plasma membrane and enriched in detergent insoluble microdomains. In the electron microscopy studies, Sulph I staining was also associated with mitochondria and villi structures. In conclusion, RIN-38 cells might be an appropriate model, as a complement to isolated islets where the amount of material often limits the experiments, to further explore the role of sulfatide in insulin secretion and signal transduction of beta cells.
以往使用各种哺乳动物胰腺以及大鼠胰腺新鲜分离胰岛的研究提供了证据,支持糖鞘脂硫苷脂可能参与胰岛素加工和分泌。在本研究中,对常用于β细胞功能研究模型的β细胞系RINr1046 - 38(RIN - 38)中的硫苷脂表达和代谢进行了研究,并与新鲜分离胰岛的先前研究结果进行了比较。RIN - 38细胞表达的硫苷脂量与分离的大鼠胰岛相似(2.7±1.1 nmol/mg蛋白质,n = 19),并且也遵循相同的代谢途径,主要是通过循环利用。此外,与分离胰岛中的发现一致,从RIN - 38细胞中分离出的硫苷脂主要种类含有C16:0和C24:0脂肪酸。通过应用亚细胞分离、电子显微镜以及免疫细胞化学技术,显示硫苷脂定位于分泌颗粒、质膜,并富集于去污剂不溶性微区。在电子显微镜研究中,Sulph I染色也与线粒体和绒毛结构相关。总之,RIN - 38细胞可能是一个合适的模型,作为对分离胰岛的补充,因为分离胰岛的材料量常常限制实验,以进一步探索硫苷脂在β细胞胰岛素分泌和信号转导中的作用。