Insights into the Nanobiology of Growth Hormone Secretion

The fact that partially empty vesicles are generated following cell secretion suggested that secretory vesicles do not collapse at the cell plasma membrane but, rather, transiently dock and fuse at the plasma membrane to expel a portion of their contents before retracting or undergoing endocytosis into the cell. Such a process has also been referred to in the literature as a "kiss-and-run" mechanism. This mechanism of cell secretion was conclusively demonstrated following the discovery of permanent cup-shaped lipoprotein structures at the cell plasma membrane, called "porosomes", where secretory vesicles transiently dock and fuse to expel intravesicular contents from the cell. Porosomes are present in all secretory cells, from the digestive enzyme-secreting pancreatic acinar cells, to the hormone-releasing growth hormone cells, mast cells, chromaffin cells, hair cells of the inner ear, to neurons secreting neurotransmitters. Hence, it can be asserted that porosomes are the universal secretory machinery in the plasma membrane of secretory cells. Therefore, the discovery of the porosome has resulted in a paradigm shift in our understanding of cell secretion. Rapid transport of secretory vesicles containing hormones to the plasma membrane is powered by high-energy molecules such as ATP, GTP or NADH. Immunogold labeled transmission electron microscopy (TEM) was used to determine the total number of secretory vesicles in resting and in GH-stimulated porcine pituitary cells. We identified three categories of vesicles: filled, empty, and partly empty. Resting GH cells contained more than twice as many filled vesicles than did the stimulated ones. However, stimulated cells contained nearly twice as many empty vesicles and 2.5 times more partly empty vesicles than did resting cells. Secretory vesicles in GH cells further revealed the localization of GH only in electron dense vesicles in both resting and stimulated cells. No change in the total number of secretory vesicles following secretion was observed. These results are consistent with a mechanism that, after stimulation of secretion, vesicles transiently dock and fuse at the porosome to establish a fusion pore, through which intravesicular contents are released.