D-Glucose uptake in fish hepatocytes: mediated by transporter in rainbow trout ( Oncorhynchus mykiss), but only by diffusion in prespawning lamprey ( Lampetra fluviatilis) and in RTH-149 cell line

Abstract

The transport of d-glucose into rainbow trout ( Oncorhynchus mykiss) and river lamprey ( Lampetra fluviatilis) hepatocytes, as well as into rainbow trout hepatoblastoma cell line RTH-149 was studied using tracer methods. The half-time for d-glucose equilibration was 15 s for rainbow trout. The half-times for the non-metabolizable d-glucose analog, 3- O-methyl- d-glucose equilibration were 8 s, 37 s and 38 s for rainbow trout, lamprey and RTH-149 cells, respectively. The 3- O-methyl- d-glucose was taken up by rainbow trout hepatocytes by facilitated diffusion in addition to simple diffusion. The uptake showed saturation kinetics with the K m of 37 mM and V max of 62 mmol kg −1 cells min −1. The uptake was sensitive to phloretin and cytochalasin B, but not affected by ouabain. The 3- O-methyl- d-glucose uptake by lamprey hepatocytes and RTH-149 cells showed no indication of saturation up to 160 mM, and was not affected by phloretin, cytochalasin B or ouabain, which suggests the mode of transport to be by passive diffusion. However, immunocytochemical stainings revealed the existence of mammalian type GLUT1 and GLUT2 transporters in all cells studied. The lack of a functioning carrier-mediated glucose uptake in lamprey hepatocytes might be due to its physiological state (prespawning starvation). The minor 3- O-methyl- d-glucose uptake into RTH-149 cells compared to freshly isolated rainbow trout hepatocytes might reflect low metabolic activity of the cell lines. Under the conditions applied the RTH-149 cell line is no suitable in vitro model for glucose transport in fish cells.