Cytoplasmic dynein participates in apically targeted stimulated secretory traffic in primary rabbit lacrimal acinar epithelial cells.
A major function of the acinar cells of the lacrimal gland is the production and stimulated release of tear proteins into ocular surface fluid. We investigate the participation of cytoplasmic dynein in carbachol-stimulated traffic to the apical plasma membrane in primary rabbit lacrimal acinar epithelial cells. Confocal fluorescence microscopy revealed a major carbachol-induced, microtubule-dependent recruitment of cytoplasmic dynein and the dynactin complex into the subapical region. Colocalization studies, sorbitol density gradient/phase partitioning analysis and microtubule-affinity purification of membranes showed that some dynein and dynactin complex were associated with VAMP2-enriched membranes. Adenovirus-mediated overexpression of p50/dynamitin inhibited the recruitment and colocalization of dynein, the dynactin complex and VAMP2 in the subapical region. Nocodazole treatment and p50/dynamitin overexpression also depleted subapical stores of rab3D in resting acini, suggesting that dynein activity was also involved in maintenance of rab3D-enriched secretory vesicles. These data implicate cytoplasmic dynein in stimulated traffic to the apical plasma membrane in these secretory epithelial cells.
Wang
Yanru
Y
Department of Pharmaceutical Sciences, University of Southern California, 1985 Zonal Avenue, Los Angeles, CA 90033, USA.
Jerdeva
Galina
G
Yarber
Francie A
FA
da Costa
Silvia R
SR
Xie
Jiansong
J
Qian
Limin
L
Rose
Chadron M
CM
Mazurek
Constance
C
Kasahara
Noriyuki
N
Mircheff
Austin K
AK
Hamm-Alvarez
Sarah F
SF
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002217:Carbachol / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D000072159:Dynactin Complex; D004398:Dyneins / Q000737:chemistry / Q000502:physiology*; D004847:Epithelial Cells / Q000378:metabolism*; D005089:Exocytosis; D005260:Female; D007765:Lacrimal Apparatus / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000737:chemistry; D008870:Microtubules / Q000378:metabolism; D015739:Nocodazole / Q000494:pharmacology; D050683:R-SNARE Proteins; D011817:Rabbits; D013347:Subcellular Fractions / Q000378:metabolism; D020692:rab3 GTP-Binding Proteins / Q000378:metabolism
D003902:Detergents; D000072159:Dynactin Complex; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D050683:R-SNARE Proteins; D002217:Carbachol; D004398:Dyneins; D020692:rab3 GTP-Binding Proteins; D015739:Nocodazole