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© The Rockefeller University Press, 0021-9525/1998//135 $5.00
The Journal of Cell Biology, Volume 141, Number 1, , 1998 135-142

Regular Articles

Apical Vesicles Bearing Inositol 1,4,5-trisphosphate Receptors in the Ca2+Initiation Site of Ductal Epithelium of Submandibular Gland



Miki Yamamoto-Hino*,{ddagger}, Atsushi Miyawaki{ddagger}, Akihisa Segawa§, Eijiro Adachi§, Shohei Yamashina§, Toyoshi Fujimoto||, Tomoyasu Sugiyama, Teiichi Furuichi{ddagger}, Mamoru Hasegawa, and Katsuhiko Mikoshiba*,{ddagger}

* Developmental Neurobiology Laboratory, RIKEN Brain Science Institute, Wako-City, Saitama 351, Japan; {ddagger} Department of Molecular Neurobiology, The Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108, Japan; § Department of Anatomy and Cell Biology, Faculty of Medicine, Kitasato University, Sagamihara-shi, Kanagawa 228, Japan; || Department of Anatomy, School of Medicine, Gunma University, Maebashi-shi, Gunma 371, Japan; and Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Machida-shi, Tokyo 194, Japan

In polarized epithelial cells, agonists trigger Ca2+ waves and oscillations. These patterns may be caused by the compartmentalization of inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ pools into specific regions. We have investigated the relationship between the distribution of IP3 receptors (IP3Rs) and the spatiotemporal pattern of Ca2+ signaling in the duct cells of the rat submandibular gland (SMG). Using immunofluorescence, although labeling was somewhat heterogeneous, the IP3Rs were colocalized to the apical pole of the duct cells. Immunoelectron microscopy identified small apical vesicles bearing IP3R2 in some types of duct cells. Real-time confocal imaging of intact ducts demonstrated that, after carbachol stimulation, an initial Ca2+ spike occurred in the apical region. Subsequently, repetitive Ca2+ spikes spread from the apical to the middle cytoplasm. These apical Ca2+ initiation sites were found only in some "pioneer cells," rather than in all duct cells. We performed both Ca2+ imaging and immunofluorescence on the same ducts and detected the strongest immunosignals of IP3R2 in the Ca2+ initiation sites of the pioneer cells. The subcellular localization and expression level of IP3Rs correlated strongly with the spatiotemporal nature of the intracellular Ca2+ signal and distinct Ca2+ responses among the rat SMG duct cells.

Abbreviations used in this paper: [Ca2+]i, intracellular calcium concentration; IP3, inositol 1,4,5-trisphosphate; IP3R, IP3 receptor; PSS, physiological salt solution; SMG, submandibular gland.

Address all correspondence to Miki Yamamoto-Hino, Department of Molecular Neurobiology, The Institute of Medical Science, The University of Tokyo, Tokyo 108, Japan. Tel.: (81) 3-5449-5319. Fax: (81) 3-5449-5420.


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