Receptors for neuroligands in the paranodal Schwann cell region of a myelinated nerve fiber could have important functions. We have used confocal laser scanning microscopy in combination with Ca(2+)-sensitive fluorescent dyes to study the possible effects of purinergic agonists on the free intracellular calcium concentration ([Ca2+]i) in paranodes of isolated rat spinal roots. Application of ATP in concentrations of 100 and 300 microM resulted in a transient rise in [Ca2+]i in about 57% of the paranodal Schwann cell regions studied. UTP was equipotent to ATP whereas adenosine, beta,gamma-methylene ATP, and elevation of the extracellular K+ concentration by 10 mM had no effect on [Ca2+]i. These data indicate the presence of the P2Y2 (previously termed P2U) subtype of P2 receptors in the paranodal Schwann cell membrane of rat spinal root myelinated nerve fibers.