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Vol. 17, Issue 5, 2424-2438, May 2006

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Motion Matters: Secretory Granule Motion Adjacent to the Plasma Membrane and Exocytosis

Miriam W. Allersma ^* , Mary A. Bittner ^*, Daniel Axelrod , and Ronald W. Holz ^*

^* Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109-0632; Biophysics Research Division, Department of Physics, University of Michigan, Ann Arbor, MI 48109-0632

Submitted October 11, 2005; Revised February 15, 2006; Accepted February 22, 2006
Monitoring Editor: Jennifer Lippincott-Schwartz

Total internal reflection fluorescence microscopy was used to monitor changes in individual granule motions related to the secretory response in chromaffin cells. Because the motions of granules are very small (tens of nanometers), instrumental noise in the quantitation of granule motion was taken into account. ATP and Ca²⁺, both of which prime secretion before fusion, also affect granule motion. Removal of ATP in permeabilized cells causes average granule motion to decrease. Nicotinic stimulation causes a calcium-dependent increase in average granule motion. This effect is more pronounced for granules that undergo exocytosis than for those that do not. Fusion is not preceded by a reduction in mobility. Granules sometimes move 100 nm or more up to and within a tenth of a second before fusion. Thus, the jittering motion of granules adjacent to the plasma membrane is regulated by factors that regulate secretion and may play a role in secretion. Motion continues until shortly before fusion, suggesting that interaction of granule and plasma membrane proteins is transient. Disruption of actin dynamics did not significantly alter granule motion.

Abbreviations used: DMPP, 1,1 dimethyl-4-phenylpiperzinium; GFP, green fluorescent protein; NaGEP, sodium glutamate solution with EGTA and PIPES; PSS, physiological salt solution; R motion, motion in the plane of the coverslip (x,y); TIRFM, total internal reflection fluorescence microscopy.

Address correspondence to: Miriam W. Allersma (allersma{at}umich.edu).

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