TY - JOUR
T1 - Membrane-specific expression of functional purinergic receptors in normal human nasal epithelial cells
AU - Kim, Chang Hoon
AU - Kim, Sung Shik
AU - Choi, Jae Young
AU - Shin, Ji Hyun
AU - Kim, Jin Young
AU - Namkung, Wan
AU - Lee, Jeung Gweon
AU - Lee, Min Goo
AU - Yoon, Joo Heon
PY - 2004/10/1
Y1 - 2004/10/1
N2 - Extracellular purines and pyrimidines regulate various physiological responses via the cell surface receptors known as purinoreceptors and may exert autocrine or paracrine effects on ion transport, fluid transport, ciliary beat frequency, and mucin secretion. Therefore, this study aims to investigate the expression patterns of the purinoreceptors in normal human nasal epithelial (NHNE) cells, In RT-PCR, the mRNAs for several P2X (P2X3, P2X 4, P2X7) and P2Y (P2Y1, P2Y2, (P2Y4, P2Y6, P2Y11, P2Y12) receptors were identified in NHNE cells. Functional localizations of P2 receptors were investigated by measuring intracellular calcium concentration ([Ca 2+]i) increases in membrane-specific manner using a double-perfusion chamber. Absence of the responses of αβ-methylene ATP and 2-methylthio-ATP excluded functionally active P2X3, P2X4, and P2Y1 receptors as far as [Ca2+]i increase is concerned. Applications with ATP and UTP revealed that luminal membranes of NHNE cells express P2Y2 and P2Y6 receptors and basolateral membranes express P2Y2 receptor. Expressions of P2Y2 and P2Y6 receptors in NHNE cells were further verified by immunoblotting using specific antibodies. In addition, the results with 2,3-O-(4-benzoyl)- benzoyl-ATP indicate that the P2Y11 receptor may be present on the luminal side. In conclusion, the NHNE cells express functionally active P2Y 2, P2Y6, and P2Y11 receptors in a membrane-specific pattern, which may play an important role in the control of mucin and fluid secretion in NHNE cells.
AB - Extracellular purines and pyrimidines regulate various physiological responses via the cell surface receptors known as purinoreceptors and may exert autocrine or paracrine effects on ion transport, fluid transport, ciliary beat frequency, and mucin secretion. Therefore, this study aims to investigate the expression patterns of the purinoreceptors in normal human nasal epithelial (NHNE) cells, In RT-PCR, the mRNAs for several P2X (P2X3, P2X 4, P2X7) and P2Y (P2Y1, P2Y2, (P2Y4, P2Y6, P2Y11, P2Y12) receptors were identified in NHNE cells. Functional localizations of P2 receptors were investigated by measuring intracellular calcium concentration ([Ca 2+]i) increases in membrane-specific manner using a double-perfusion chamber. Absence of the responses of αβ-methylene ATP and 2-methylthio-ATP excluded functionally active P2X3, P2X4, and P2Y1 receptors as far as [Ca2+]i increase is concerned. Applications with ATP and UTP revealed that luminal membranes of NHNE cells express P2Y2 and P2Y6 receptors and basolateral membranes express P2Y2 receptor. Expressions of P2Y2 and P2Y6 receptors in NHNE cells were further verified by immunoblotting using specific antibodies. In addition, the results with 2,3-O-(4-benzoyl)- benzoyl-ATP indicate that the P2Y11 receptor may be present on the luminal side. In conclusion, the NHNE cells express functionally active P2Y 2, P2Y6, and P2Y11 receptors in a membrane-specific pattern, which may play an important role in the control of mucin and fluid secretion in NHNE cells.
UR - http://www.scopus.com/inward/record.url?scp=4544377752&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=4544377752&partnerID=8YFLogxK
U2 - 10.1152/ajplung.00285.2003
DO - 10.1152/ajplung.00285.2003
M3 - Article
C2 - 15208093
AN - SCOPUS:4544377752
VL - 287
SP - L835-L842
JO - American Journal of Physiology - Lung Cellular and Molecular Physiology
JF - American Journal of Physiology - Lung Cellular and Molecular Physiology
SN - 1040-0605
IS - 4 31-4
ER -