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© The Rockefeller University Press, 0021-9525/1997//497 $5.00
The Journal of Cell Biology, Volume 139, Number 2, , 1997 497-506

Article

ATP- and Gap Junction–dependent Intercellular Calcium Signaling in Osteoblastic Cells



Niklas R. Jørgensen*,{ddagger}, Steven T. Geist*, Roberto Civitelli*, and Thomas H. Steinberg*

* Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110; and {ddagger} Osteoporosis Research Center, DK-1399 Copenhagen, Denmark

Many cells coordinate their activities by transmitting rises in intracellular calcium from cell to cell. In nonexcitable cells, there are currently two models for intercellular calcium wave propagation, both of which involve release of inositol trisphosphate (IP3)- sensitive intracellular calcium stores. In one model, IP3 traverses gap junctions and initiates the release of intracellular calcium stores in neighboring cells. Alternatively, calcium waves may be mediated not by gap junctional communication, but rather by autocrine activity of secreted ATP on P2 purinergic receptors. We studied mechanically induced calcium waves in two rat osteosarcoma cell lines that differ in the gap junction proteins they express, in their ability to pass microinjected dye from cell to cell, and in their expression of P2Y2 (P2U) purinergic receptors. ROS 17/2.8 cells, which express the gap junction protein connexin43 (Cx43), are well dye coupled, and lack P2U receptors, transmitted slow gap junction-dependent calcium waves that did not require release of intracellular calcium stores. UMR 106-01 cells predominantly express the gap junction protein connexin 45 (Cx45), are poorly dye coupled, and express P2U receptors; they propagated fast calcium waves that required release of intracellular calcium stores and activation of P2U purinergic receptors, but not gap junctional communication. ROS/P2U transfectants and UMR/Cx43 transfectants expressed both types of calcium waves. Gap junction–independent, ATP-dependent intercellular calcium waves were also seen in hamster tracheal epithelia cells. These studies demonstrate that activation of P2U purinergic receptors can propagate intercellular calcium, and describe a novel Cx43-dependent mechanism for calcium wave propagation that does not require release of intracellular calcium stores by IP3. These studies suggest that gap junction communication mediated by either Cx43 or Cx45 does not allow passage of IP3 well enough to elicit release of intracellular calcium stores in neighboring cells.

Abbreviations used in this paper: Cx43 and Cx45, connexin 43 and 45; HTE, hamster tracheal epithelia; IP3, inositol trisphosphate.

Address all correspondence to Thomas H. Steinberg, Washington University School of Medicine, Infectious Diseases, Box 8051, 660 South Euclid Avenue, St. Louis, MO 63110. Tel.: (314) 362-9218. Fax: (314) 362-9218. E-mail: steinber{at}id.wustl.edu

N.R. Jørgensen's current address is Osteoporosis Research Center, Copenhagen Municipal Hospital, Oster Farimagsgade 5, DK-1399 Copenhagen K, Denmark.


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