Changes in kinesin distribution and phosphorylation occur during regulated secretion in pancreatic acinar cells

Kimberly J. Marlowe, Parvaiz Farshori, Rochelle R. Torgerson, Karen L. Anderson, Laurence J Miller, Mark A Mc Niven

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

In secretory cells, microtubule- (Mt-) based motor enzymes are thought to support transport of secretory vesicles to the cell surface for subsequent release. At present, the role of Mts and kinesin in secretory vesicle transport in exocrine epithelial cells has not been defined. Furthermore, it is unclear whether an agonist-induced secretory event modifies kinesin function and distribution, thus altering vesicle transport. To this end, we utilized isolated rat pancreatic acini and cultured rat pancreatic acinar cells to examine the role of Mts and kinesin in regulated secretion. Exposure of cells to cytoskeletal antagonistic drugs demonstrated that the observed movements of apically clustered zymogen granules (ZGs) are supported by Mts, but not actin. Morphological studies of Mt organization in polarized acini show that Mt plus ends extend outward from the apical membrane toward the cell center. Immunofluorescence microscopy in both cell models revealed a clear association of kinesin with apical ZGs, while quantitative immunoblot analysis of pancreatic subcellular fractions confirmed kinesin enrichment on ZG membranes. In addition, microinjection of kinesin antibodies into cultured acinar cells inhibited ZG movements. Indirect immunofluorescence staining of isolated cells and quantitative Western blotting of isolated ZGs revealed that kinesin association with granule membranes increased up to 3-fold in response to a secretory stimulus. Autoradiographic studies of 32P-labeled acini showed up to a 6-fold increase in kinesin heavy chain (KHC) phosphorylation during stimulated secretion. These studies provide the first direct evidence that Mts and kinesin support ZG movements and that physiological agonists induce a marked phosphorylation of KHC while increasing the association of kinesin with ZG membranes. These changes during agonist stimulation suggest that the participation of kinesin in zymogen secretion is regulated.

Original languageEnglish (US)
Pages (from-to)140-152
Number of pages13
JournalEuropean Journal of Cell Biology
Volume75
Issue number2
StatePublished - 1998

Fingerprint

Kinesin
Acinar Cells
Secretory Vesicles
Phosphorylation
Transport Vesicles
Membranes
Enzyme Precursors
Subcellular Fractions
Microinjections
Indirect Fluorescent Antibody Technique
Fluorescence Microscopy
Microtubules
Actins
Cultured Cells

Keywords

  • Kinesin
  • Microtubules
  • Regulated secretion
  • Zymogen granule movement

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Changes in kinesin distribution and phosphorylation occur during regulated secretion in pancreatic acinar cells. / Marlowe, Kimberly J.; Farshori, Parvaiz; Torgerson, Rochelle R.; Anderson, Karen L.; Miller, Laurence J; Mc Niven, Mark A.

In: European Journal of Cell Biology, Vol. 75, No. 2, 1998, p. 140-152.

Research output: Contribution to journalArticle

Marlowe, Kimberly J. ; Farshori, Parvaiz ; Torgerson, Rochelle R. ; Anderson, Karen L. ; Miller, Laurence J ; Mc Niven, Mark A. / Changes in kinesin distribution and phosphorylation occur during regulated secretion in pancreatic acinar cells. In: European Journal of Cell Biology. 1998 ; Vol. 75, No. 2. pp. 140-152.
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