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Biophysical Journal 84:4127-4134 (2003)
© 2003 The Biophysical Society

Unexpected Mobility Variation among Individual Secretory Vesicles Produces an Apparent Refractory Neuropeptide Pool

Yuen-Keng Ng *, Xinghua Lu *, Alexandra Gulacsi *, Weiping Han *, Michael J. Saxton {dagger} and Edwin S. Levitan *

* Department of Pharmacology, E1351 Biomedical Science Tower, University of Pittsburgh, Pittsburgh, Pennsylvania 15261; and {dagger} Institute of Theoretical Dynamics, University of California, Davis, California

Correspondence: Address reprint requests to Edwin S. Levitan, Tel.: 412-648-9486; Fax: 412-648-1945; E-mail: Levitan{at}server.pharm.pitt.edu.

Most stored neuropeptide cannot be released from nerve terminals suggesting the existence of a refractory pool of dense core vesicles (DCVs). Past fluorescence photobleaching recovery, single particle tracking and release experiments suggested that the refractory neuropeptide pool corresponds to a distinct immobile fraction of cytoplasmic DCVs. However, tracking of hundreds of individual green fluorescent protein-labeled neuropeptidergic vesicles by wide-field or evanescent-wave microscopy shows that a separate immobile fraction is not evident. Instead, the DCV diffusion coefficient (D) distribution is unusually broad and asymmetric. Furthermore, the distribution shifts with a release facilitator. This unexpected variation, which could reflect heterogeneity among vesicles or in their medium, is shown to generate the appearance of a regulated refractory neuropeptide pool.




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