Regulator of G-Protein Signaling Protein 2 Modulates Purinergic Calcium and Ciliary Beat Frequency Responses in Airway Epithelia

In ciliated airway epithelial cells, purinergic stimulation increases both intracellular calcium ([Ca(2+)](i)) and ciliary beat frequency (CBF). Because regulator of G-protein signaling protein 2 (RGS2) terminates Galphaq-mediated phospholipase C activation, we examined its role in regulating purinergic signaling in human and ovine airway epithelial cells. RT-PCR of both human and ovine epithelial cell RNA yielded fragments of expected size ( approximately 491 bp) and sequence, confirming RGS2 message. Immunofluorescence demonstrated RGS2 protein expression in cultured airway epithelial cells of both species. Overexpression of an EGFP-RGS2 fusion protein (increasing RGS2 protein levels 1.8 times control, n = 28 cells) resulted in a reduced [Ca(2+)](i) and CBF response to 10 micro M ATP (human: 58 +/- 9% and 49 +/- 8% lower, respectively; n = 8 measurements, 4 cells; ovine: 56 +/- 12% and 53 +/- 16% lower, respectively; n = 5 measurements, 4 cells). Reducing RGS2 protein levels using antisense oligonucleotides increased the response of both [Ca(2+)](i) and CBF to ATP in human cells by 57 +/- 10% and 47 +/- 11%, respectively (n = 10 measurements, 6 cells), and in ovine cells by 88 +/- 13% and 48 +/- 9%, respectively (n = 10 measurements, 5 cells). These data provide functional evidence that RGS2 modulates purinergic signaling in human and ovine ciliated airway epithelial cells.