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Rab3D regulates amylase levels, not agonist-induced amylase release, in AR42J cells

Cellular & Molecular Biology Letters volume 17, pages 258–273 (2012)Cite this article

Abstract

Rab3D is a low molecular weight GTP-binding protein that associates with secretory granules in exocrine cells. AR42J cells are derived from rat pancreatic exocrine tumor cells and develop an acinar cell-like phenotype when treated with dexamethasone (Dex). In the present study, we examined the role of Rab3D in Dex-treated AR42J cells. Rab3D expression and localization were analyzed by subcellular fractionation and immunoblotting. The role of Rab3D was examined by overexpressing myc-labeled wild-type-Rab3D and a constitutively active form of Rab3D (Rab3D-Q81L) in AR42J cells. We found that Rab3D is predominantly membrane-associated in AR42J cells and co-localizes with zymogen granules (ZG). Following CCK-8-induced exocytosis, amylase-positive ZGs appeared to move towards the periphery of the cell and co-localization between Rab3D and amylase was less complete when compared to basal conditions. Overexpression of WT, but not mutant Rab3D, resulted in an increase in cellular amylase levels. Overexpression of mutant and WT Rab3D did not affect granule morphology, CCK-8-induced secretion, long-term (48 hr) basal amylase release or granule density. We conclude that Rab3D is not involved in agonist-induced exocytosis in AR42J cells. Instead, Rab3D may regulate amylase content in these cells.

Abbreviations

Dex:

dexamethasone

CCK-8:

cholecystokinin octapeptide

TGN:

Trans Golgi network

ZG:

zymogen granules

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Authors and Affiliations

  1. Hunter College, School of Health Professions, Medical Laboratory Sciences Program, Box 617, 425 East 25th Street, New York, NY, 10010, USA

    Saima Limi & Robert Raffaniello

  2. Department of Cell Biology, SUNY-Downstate Medical Center, Brooklyn, New York, USA

    George Ojakian

Authors
  1. Saima Limi
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  2. George Ojakian
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  3. Robert Raffaniello
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Correspondence to Robert Raffaniello.

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Limi, S., Ojakian, G. & Raffaniello, R. Rab3D regulates amylase levels, not agonist-induced amylase release, in AR42J cells. Cell Mol Biol Lett 17, 258–273 (2012). https://doi.org/10.2478/s11658-012-0008-5

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Cellular & Molecular Biology Letters

ISSN: 1689-1392

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