HETEROGENEOUS RESPONSES OF CELL Ca2+ IN HUMAN AIRWAY EPITHELIUM

• Published in: Experimental lung research 1999-06, Vol.25 (4), p.277-290
• Main Authors: PARADISO, A. M, BROWN, H. A, HONG YE, HARDEN, T. K, BOUCHER, R. C
• Format: Article
• Language: English
• Subjects: Adenosine 5'-triphosphate; Bradykinin; Cytokeratins; Fura-2; Histamine; Microfluorimetry; Thapsigargin; Adenosine Triphosphate - pharmacology; Adult; Air breathing; Biological and medical sciences; Bradykinin - pharmacology; Calcium - agonists; Calcium - metabolism; Calcium-Transporting ATPases - antagonists & inhibitors; Cells, Cultured; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Female; Fluorescent Antibody Technique, Indirect; Fundamental and applied biological sciences. Psychology; Fura-2 - metabolism; Histamine - pharmacology; Humans; Keratins - metabolism; Male; Nasal Mucosa - cytology; Nasal Mucosa - drug effects; Nasal Mucosa - metabolism; Receptors, Purinergic P2 - metabolism; Respiratory system: anatomy, metabolism, gas exchange, ventilatory mechanics, respiratory hemodynamics; Thapsigargin - pharmacology; Vertebrates: respiratory system
• ISSN: 0190-2148; 1521-0499
• DOI: 10.1080/019021499270196

The Ca2+-mobilizing actions of adenosine 5'-triphosphate (ATP), bradykinin, and histamine were compared in phenotypically distinct human nasal epithelial (HNE) cell types and as a function of time in cell culture. Single-cell measurements of intracellular free Ca2+ (Ca2+i, Fura-2 fluorescence) were recorded in ciliated cells 1-2 days in primary culture, and in nonciliated cells 1-2 days (keratin 14-positive) or 4-5 days (keratin 18-positive) after seeding. No difference in basal Ca2+i was noted between ciliated and nonciliated cell preparations. For ciliated and nonciliated cells studied 1-2 days in culture, ATP, bradykinin, and histamine elicited a cytosolic Ca2+ response in 100 % of the cells examined. For nonciliated HNE cells maintained 4-5 days in culture, ATP (10-4 M) increased cytosolic Ca2+ in all cells tested, but only 85 % of the cells responded to bradykinin (10-5 M) addition, and 65 % to histamine (10-4 M) stimulation. In terms of the absolute change of Ca2+i (Delta Ca2+i, peak-basal value), the efficacy was ATP > bradykinin > histamine for the 3 HNE cell preparations. However, the Delta Ca2+i in response to agonists was smaller in nonciliated HNE cells studied 1-2 days or 4-5 days in culture as compared to the ciliated cell preparation. Thapsigargin (300 nM), an agent that mobilizes Ca2+i, was equally effective in raising cytosolic Ca2+ in nonciliated (1 - 2 days and 4-5 days in culture) and ciliated HNE cells. These data show that ciliated cells consistently respond to all agonists, whereas the cytosolic Ca2+ response to ATP, bradykinin, and histamine in nonciliated cells was quantitatively reduced at a comparable time period (1-2 days) and became smaller and less frequent in nonciliated cell preparations maintained 4-5 days in culture. These results demonstrate time-dependent differences in the magnitude and frequency of cytosolic Ca2+responses to certain agonists, strongly indicating that measurements of Ca2+ in HNE cells must account for the heteri ogeneity of the cell types and the time cells are maintained in primary culture.