Schwann cell ATP‐mediated calcium increases in vitro and in situ are dependent on contact with neurons

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Abstract
Schwann cells freshly isolated from the sciatic nerves of neonatal rats respond to exogenously applied ATP with a rapid increase in cytosolic calcium. This increase in [Ca2+]i is mediated by a P2Y‐purinergic pathway (Lyons et al.: J. Neurochem. 63:552–560, 1994) and was measured using the calcium indicator dye, fura‐2/AM, and a video‐enhanced calcium imaging system. The ability to respond to ATP with increases in intracellular calcium is lost over a period of several days in culture; this loss can be prevented or reversed by application of cAMP analogs in a defined medium. We now demonstrate that the direct contact of Schwann cells with neurons also induces and stabilizes this ATP responsiveness. The induction of ATP responsiveness was observed among all Schwann cells contacting neurites, including those forming myelin, and regardless of whether the source of neurons was dorsal root ganglion neurons or superior cervical ganglion neurons. Approximately 85% of Schwann cells responded to ATP over the time studied (72 d in coculture). Addition of axolemma to Schwann cell cultures did not induce ATP responsiveness. We also examined the ATP responsiveness of Schwann cells in situ (excised nerves) using laser‐scanning confocal microscopy and the calcium indicator dye, fluo‐3/AM. Schwann cells in intact sciatic nerve segments isolated from neonatal and 16–day‐old rats exhibited ATP‐mediated [Ca2+]i increases. We conclude that neuronal contact is necessary for the expression of the ATP‐mediated calcium responses in Schwann cells and that these responses are independent of myelin formation or maintenance. Copy 1995 Wiley‐Liss, Inc.