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The Journal of Neuroscience, May 15, 2002, 22(10):3890-3897

Nerve Growth Factor-Induced Differentiation Changes the Cellular Organization of Regulated Peptide Release by PC12 Cells

Yuen-Keng Ng^{1, *}, Xinghua Lu^{1, *}, Simon C. Watkins², Graham C. R. Ellis-Davies³, and Edwin S. Levitan¹

Departments of ¹ Pharmacology and ² Cell Biology and Physiology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, and ³ Department of Pharmacology and Physiology, MCP/Hahnemann University, Philadelphia, Pennsylvania 19102

PC12 cells, like endocrine chromaffin cells, undergo neuronal-like differentiation in response to nerve growth factor (NGF). Here we report that this phenotype conversion produces major changes in release of a green fluorescent protein-tagged neuropeptide-hormone. First, the spatial distribution of the releasable pool is altered; peptide release from untreated cells is supported predominantly by membrane-proximal vesicles, whereas a diffuse pool at the ends of processes is used by NGF-treated cells. Second, the time course of release evoked by photolysis of caged Ca²⁺ is faster after differentiation. High-resolution measurements suggest that a slow step before membrane fusion dominates the kinetics of release in untreated cells. Finally, the effect of actin microfilament depolymerization on total release is altered by NGF treatment. This implies that the mechanism that limits the size of the releasable pool is altered by phenotype conversion. Therefore, the cellular organization of peptide release is plastic and changes in response to NGF. This flexibility may be used to generate cell-specific release properties.

Key words: neuropeptide release; hormone release; GFP; caged calcium; actin; secretory vesicle; neuronal differentiation; releasable pool

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^* Y.-K.N. and X.L. contributed equally to this work.

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Copyright © 2002 Society for Neuroscience  0270-6474/02/22103890-08$05.00/0

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