Research Paper Volume 12, Issue 10 pp 8923—8938

Influence of human amylin on the membrane stability of rat primary hippocampal neurons

Figure 5. Scanning electron microscopy images of primary cultured hippocampal cells with or without hAmylin (10 μM, 1 h). (A) The plasma membrane was smooth and integral for primary cultured neurons and astrocytes without amylin incubation. After the cells had been treated with hAmylin (10 μM), significant plasma membrane damage was observed on the surface of primary cultured neurons and astrocytes. (B) Intracellular ROS generation induced by 10 μM hAmylin was measured in hippocampal neurons labeled with DCFH-DA dye. Representative traces are shown of the effects of 1 μM hAmylin, 10 μM hAmylin, 10 μM hAmylin + 1 μM CsA and 10 μM hAmylin + free Ca2+ on ROS generation. (C) Significant ROS generation was induced by 10 μM hAmylin, and was not inhibited by 1 μM CsA or free extracellular Ca2+. (D) Changes in neuronal DCFH-DA fluorescence before and after 10 μM hAmylin incubation. (E) The reduction in the mtΔΨ induced by 10 μM hAmylin was measured in hippocampal neurons labeled with JC-1 dye. Representative traces are shown of the effects of 1 μM hAmylin, 10 μM hAmylin, 10 μM hAmylin + 1 μM CsA and 10 μM hAmylin + free Ca2+ on the mtΔΨ. (F) Significant mtΔΨ reduction was induced by 10 μM hAmylin and inhibited by 1 μM CsA, but not by free extracellular Ca2+. (G) Changes in neuronal JC-1 fluorescence before and after 10 μM hAmylin incubation. ***p < 0.001 versus 10 μM hAmylin (one-way ANOVA followed by Bonferroni’s post hoc test).