Quantitative analysis of exocytosis directly visualized in living chromaffin cells

Abstract

Chromaffin cells isolated from the bovine adrenal medulla were observed under a Nomarski microscope through a CCD camera and an image processor. Exocytotic events of individual granules including fusion, extrusion, swelling, omega-figure formation, and membrane retrieval were visualized in individual cells stimulated by acetylcholine or K-rich solution. Initial steps were quicker than 16 ms, and the membrane retrieval was slower than 1-60 s. These findings provided a light microscopic proof for the exocytosis hypothesis as well as a basis for quantification of hormonal release. The technique was used to demonstrate significant secretion induced by a muscarinic agonist.