Vesicular roundness and compound release in PC-12 cells

The principal goals of this study were to establish a quantitative morphological analysis of spatial and regional properties of dense core vesicles, and to use this analysis to assess whether homotypic fusion is prominent in chronically treated PC-12 cells at elevated release levels. Simple computerized image processing of electron-micrographs provided the binary images of vesicular dense cores, whilst the artificial intelligence methods were needed to determine the vesicular membranes. As in the past, the presence of large, highly irregular vesicles, provided the morphological evidence of fused vesicles, but the irregularity of vesicular shape was assessed quantitatively-from its roundness. Free space of each vesicle was determined from the distance to its nearest-neighbor, or from the size of its Voronoi polygon. Within a Voronoi polygon, each point is closer to that vesicle than to any other vesicle. Large vesicles were not less round and did not have larger free space, as expected if they result from fusion of several smaller vesicles. In conclusion, we present a novel and rigorous morphological analysis of spatial and regional properties of dense core vesicles. The results demonstrate that the homotypic fusion is not prominent in PC-12 cells, before or following a chronic treatment that enhances release.