Human Reproduction, Vol. 11, No. 1, pp. 212-217, 1996
© 1996 European Society of Human Reproduction and Embryology
research-article |
Human Fallopian tubal epithelial cells in vitro:establishment of polarity and potential role of intracellular calcium and extracellular ATP in fluid secretion
Department of Biology, University of York PO Box 373, York YO1 5YW 1ICRF Cancer Medicine Research Unit, St Jamess University Hospital Beckett Street, Leeds LS9 7TF, UK
Correspondence: 2To whom correspondence should be addressed
A pure population of human Fallopian tubal epithelial cells has been isolated by enzyme digestion, grown in primary culture and used to explore the biochemical basis of oviduct fluid secretion. Confluence was achieved in 37 days. Immunocytochemical labelling for cytokeratins indicated that the cells were epithelial in nature and formed extensive desmosomal contacts, producinga polarized layer in culture. By growing the cells on collagen-impregnated filters, a small transepithelial electrical potential difference could be recorded, with the apical side of the cells negative with respect to the basal side. In addition, the consumption of glucose and the appearance of lactate were greater on the basal than on the apical side of the cells. Because intracellular Ca2+ ([Ca2+]1) is well established as a signal transduction agent in epithelial fluid secretion, the effect of a wide range of agonists on [Ca2+]1 in isolated tubal epithelial cells was studied using Fura-2. The only agent which induced a change in [Ca2+]1 was extracellular ATP. The transients induced were dependent on both intracellular and extracellular calcium. ATP added to the basal side of the cells of the polarized layer induced a transient increase in the potential difference. The data are consistent with a potential role for extracellular ATP in the regulation of human tubal fluid formation.
Key words: epithelia/Fallopian tube/human tubal fluid/intracellular calcium/purinergic receptors
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