HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplemental Videos All Versions of this Article: 289/4/C1052    most recent 00546.2004v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (10) Citing Articles via Google Scholar Google Scholar Articles by Jerdeva, G. V. Articles by Hamm-Alvarez, S. F. Search for Related Content PubMed PubMed Citation Articles by Jerdeva, G. V. Articles by Hamm-Alvarez, S. F.

CALL FOR PAPERS
Protein and Vesicle Trafficking, Cytoskeleton

Dominant-negative PKC- impairs apical actin remodeling in parallel with inhibition of carbachol-stimulated secretion in rabbit lacrimal acini

Departments of ¹Pharmaceutical Sciences, ²Ophthalmology, and ³Physiology and Biophysics, University of Southern California, Los Angeles, California; ⁴Pacific Northwest Research Institute, Seattle, Washington; and ⁵Schepens Eye Research Institute, Boston, Massachusetts

Submitted 9 November 2004 ; accepted in final form 25 May 2005

We investigated the involvement of PKC- in apical actin remodeling in carbachol-stimulated exocytosis in reconstituted rabbit lacrimal acinar cells. Lacrimal acinar PKC- cosedimented with actin filaments in an actin filament binding assay. Stimulation of acini with carbachol (100 µM, 2–15 min) significantly (P 0.05) increased PKC- recovery with actin filaments in two distinct biochemical assays, and confocal fluorescence microscopy showed a significant increase in PKC- association with apical actin in stimulated acini as evidenced by quantitative colocalization analysis. Overexpression of dominant-negative (DN) PKC- in lacrimal acini with replication-defective adenovirus (Ad) resulted in profound alterations in apical and basolateral actin filaments while significantly inhibiting carbachol-stimulated secretion of bulk protein and -hexosaminidase. The chemical inhibitor GF-109203X (10 µM, 3 h), which inhibits PKC-, -, -, and -, also elicited more potent inhibition of carbachol-stimulated secretion relative to Gö-6976 (10 µM, 3 h), which inhibits only PKC- and -. Transduction of lacrimal acini with Ad encoding syncollin-green fluorescent protein (GFP) resulted in labeling of secretory vesicles that were discharged in response to carbachol stimulation, whereas cotransduction of acini with Ad-DN-PKC- significantly inhibited carbachol-stimulated release of syncollin-GFP. Carbachol also increased the recovery of secretory component in culture medium, whereas Ad-DN-PKC- transduction suppressed its carbachol-stimulated release. We propose that DN-PKC- alters lacrimal acinar apical actin remodeling, leading to inhibition of stimulated exocytosis and transcytosis.

lacrimal gland; acinar epithelial cell; exocytosis; polymeric immunoglobulin A receptor

This article has been cited by other articles:

				M. L. McDonald, Y. Wang, S. Selvam, T. Nakamura, R. H. Chow, J. E. Schechter, S. C. Yiu, and A. K. Mircheff Cytopathology and Exocrine Dysfunction Induced in Ex Vivo Rabbit Lacrimal Gland Acinar Cell Models by Chronic Exposure to Histamine or Serotonin Invest. Ophthalmol. Vis. Sci., July 1, 2009; 50(7): 3164 - 3175. [Abstract] [Full Text] [PDF]

				R. R. Marchelletta, D. T. Jacobs, J. E. Schechter, R. E. Cheney, and S. F. Hamm-Alvarez The class V myosin motor, myosin 5c, localizes to mature secretory vesicles and facilitates exocytosis in lacrimal acini Am J Physiol Cell Physiol, July 1, 2008; 295(1): C13 - C28. [Abstract] [Full Text] [PDF]

				E. Evans, W. Zhang, G. Jerdeva, C.-Y. Chen, X. Chen, S. F. Hamm-Alvarez, and C. T. Okamoto Direct interaction between Rab3D and the polymeric immunoglobulin receptor and trafficking through regulated secretory vesicles in lacrimal gland acinar cells Am J Physiol Cell Physiol, March 1, 2008; 294(3): C662 - C674. [Abstract] [Full Text] [PDF]

				C. Ehre, Y. Zhu, L. H. Abdullah, J. Olsen, K. I. Nakayama, K. Nakayama, R. O. Messing, and C. W. Davis nPKC{varepsilon}, a P2Y2-R downstream effector in regulated mucin secretion from airway goblet cells Am J Physiol Cell Physiol, November 1, 2007; 293(5): C1445 - C1454. [Abstract] [Full Text] [PDF]

				Y. Wang, C. T. Chiu, T. Nakamura, A. M. Walker, B. Petridou, M. D. Trousdale, S. F. Hamm-Alvarez, J. E. Schechter, and A. K. Mircheff Elevated prolactin redirects secretory vesicle traffic in rabbit lacrimal acinar cells Am J Physiol Endocrinol Metab, April 1, 2007; 292(4): E1122 - E1134. [Abstract] [Full Text] [PDF]

				J. Xie, L. Chiang, J. Contreras, K. Wu, J. A. Garner, L. Medina-Kauwe, and S. F. Hamm-Alvarez Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in Lacrimal Acini J. Virol., December 1, 2006; 80(23): 11833 - 11851. [Abstract] [Full Text] [PDF]