Disruption of the Actin Cytoskeleton Induces Fluorescent Glucose Accumulation on the Rat Hepatocytes Clone 9

Abstract

Background: Glucose transport and metabolism are highly specialized in hepatocytes. Actin cytoskeleton is fundamental to the maintenance of their morphology as well as to ensure their functionality. Here we study the effect of the actin disrupting natural compounds cytochalasin B and latrunculin A on the glucose metabolism of the Clone 9 rat hepatocytes once the glucose molecule is inside them and the effects of two hormones which main function is regulating the glucose metabolism on the actin cytoskeleton of Clone 9 cells. Methods: F-actin was labeled by using Oregon Green 514 ® phalloidin and glucose inside cells was monitored with the fluorescent D-glucose derivative; 2-NBDG. Observations and measurements were carried out by using a confocal microscope. Results: Nor insulin neither glucagon was able to induce any significant effect in the quantity of F-actin present on Clone 9 cells. But insulin triggers a strong reorganization on the pattern of distribution of F-actin. However, the actin cytoskeleton disruption induced by CB and more efficiently by Lat A caused accumulation of 2-NBDG in cells. Conclusions: These results state that disruption of the actin cytoskeleton induces fluorescent glucose accumulation on the rat hepatocytes Clone 9 suggesting that actin disrupting agents cause a blockage in the glycolytic pathway of Clone 9 hepatocytes.

Extracellular Matrix Stiffness: New Areas Affecting Cell Metabolism

“…ECM stiffness can influence glucose uptake by regulating translocation of the glucose transport protein GLUT4 to the cytomembrane ( 49 – 51 ). Besides, most glycolytic enzymes, except hexokinase that bind to mitochondria, are thought to bind to the cytoskeleton, which regulates activity of glycolytic enzymes ( 52 ). Park et al demonstrated that the actin cytoskeleton regulates glycolytic enzyme phosphofructo kinase (PFK) by limiting migration and radius of action of the E3 ubiquitin ligase tripartite motif (TRIM)-containing protein 21 (TRIM21) ( 48 ).…”

Cross-talks between c-Kit and PKC isoforms in HMC-1560 and HMC-1560,816 cells. Different role of PKCδ in each cellular line

“…In order to determine if actin cytoskeleton participates in STI571-induced nuclear PKCd translocation, latrunculin effect was studied in cytosolic and nuclear fractions. First, latrunculin, which affects actin cytoskeleton by sequestering (monomeric) G-actin [40], effect was checked in the presence of STI571. As Fig.…”