Abstract
In ciliated airway epithelial cells, purinergic stimulation increases both intracellular calcium ([Ca2+]i) and ciliary beat frequency (CBF). Because regulator of G-protein signaling protein 2 (RGS2) terminates Gαq-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 (∼ 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 [Ca2+]i and CBF response to 10 μM 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 [Ca2+]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.
| Original language | English |
|---|---|
| Pages (from-to) | 436-445 |
| Number of pages | 10 |
| Journal | American Journal of Respiratory Cell and Molecular Biology |
| Volume | 27 |
| Issue number | 4 |
| State | Published - Oct 1 2002 |
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ASJC Scopus subject areas
- Cell Biology
- Pulmonary and Respiratory Medicine
- Molecular Biology
Cite this
Regulator of G-protein signaling protein 2 modulates purinergic calcium and ciliary beat frequency responses in airway epithelia. / Nlend, Marie Christine; Bookman, Richard J.; Conner, Gregory E; Salathe, Matthias A.
In: American Journal of Respiratory Cell and Molecular Biology, Vol. 27, No. 4, 01.10.2002, p. 436-445.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Regulator of G-protein signaling protein 2 modulates purinergic calcium and ciliary beat frequency responses in airway epithelia
AU - Nlend, Marie Christine
AU - Bookman, Richard J.
AU - Conner, Gregory E
AU - Salathe, Matthias A
PY - 2002/10/1
Y1 - 2002/10/1
N2 - In ciliated airway epithelial cells, purinergic stimulation increases both intracellular calcium ([Ca2+]i) and ciliary beat frequency (CBF). Because regulator of G-protein signaling protein 2 (RGS2) terminates Gαq-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 (∼ 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 [Ca2+]i and CBF response to 10 μM 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 [Ca2+]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.
AB - In ciliated airway epithelial cells, purinergic stimulation increases both intracellular calcium ([Ca2+]i) and ciliary beat frequency (CBF). Because regulator of G-protein signaling protein 2 (RGS2) terminates Gαq-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 (∼ 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 [Ca2+]i and CBF response to 10 μM 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 [Ca2+]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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0036788754&partnerID=8YFLogxK
M3 - Article
C2 - 12356577
AN - SCOPUS:0036788754
VL - 27
SP - 436
EP - 445
JO - American Journal of Respiratory Cell and Molecular Biology
JF - American Journal of Respiratory Cell and Molecular Biology
SN - 1044-1549
IS - 4
ER -