Schwann Cells Exhibit P2Y Purinergic Receptors that Regulate Intracellular Calcium and Are Up-Regulated by Cyclic AMP Analogues

Abstract: Schwann cells establish close contact with axons during development, and this is maintained throughout life. Signaling by neurotransmitters may play an important role in Schwann cell-axon interaction. Schwann cells were examined for the presence of neuroligand receptors that are linked to...
Ausführliche Beschreibung

Abstract: Schwann cells establish close contact with axons during development, and this is maintained throughout life. Signaling by neurotransmitters may play an important role in Schwann cell-axon interaction. Schwann cells were examined for the presence of neuroligand receptors that are linked to increases in levels of cytoplasmic calcium. Schwann cell cultures were prepared from neonatal rat sciatic nerve and, after 0.25, 1, 4, 7, and 14 days in vitro (DIV), loaded with the calcium indicator dye fura 2-AM. The influence of neuroligands on the cytosolic free calcium concentration ([Ca2+]i) was then examined at each time point using a video-based imaging system. Approximately 80–95% of all freshly isolated Schwann cells responded to 10 µM ATP with a three-fold rise in [Ca2+]i. Bradykinin, glutamate, and histamine had no or only partial and inconsistent responses. The ATP-induced calcium response disappeared within 4 DIV. Culturing cells in the presence of cyclic AMP (cAMP) analogues (which induce proliferation and differentiation in vitro) restored the ability of Schwann cells to respond to ATP with increased [Ca2+]i. In the presence of cAMP analogues the extent of recovery of ATP responsiveness was dependent on serum concentration. Fifty to ninety percent of cells regained calcium responsiveness to ATP when grown in medium containing cAMP analogues and 1% serum. These cells also exhibited immunoreactivity to P0 antibody, characteristic of the myelinating lineage. In contrast, only 15–30% of the Schwann cells regained calcium responsiveness when grown in medium containing cAMP analogues and 10% serum. Under these conditions all Schwann cells exhibited immunoreactivity to antibodies against nerve growth factor receptor, characteristic of the nonmyelinating lineage, although some also contained galactocerebroside immunoreactivity. The correlation between the recovery of the ATP response and the recovery of stage-specific markers suggests that Schwann cell ATP receptor expression may be a developmental process, preferentially associated with Schwann cells moving toward the myelinating lineage. Ausführliche Beschreibung