Expression of functional gap junctions in cultured pulmonary alveolar epithelial cells

Yu Chen Lee, Clare E Yellowley-genetos, Zhongyong Li, Henry J. Donahue, D. Eugene Rannels

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Abstract

Recent observations suggest that cell-cell interactions may modulate the response of the alveolar epithelium to injury. Expression and function of gap junctions were thus evaluated in isolated alveolar type II cells. Freshly isolated (day 0) type II cells expressed mRNAs for gap junctional connexins 26, 32, and 43. Whereas connexin 26 mRNA declined ~40% in cultured cells, connexin 32 message decreased rapidly and was not detectable an day 1. In contrast, connexin 43 expression increased 10-fold by day 3 compared with day 0. Western blot confirmed a 30-fold elevation in connexin 43 protein. Connexin 45 mRNA was not detected. Functional gap junction-mediated calcium signal propagation was monitored using fura 2, a calcium-sensitive dye. Membrane deformation in a single type II cell increased intracellular calcium; the signal spread rapidly to neighboring cells by octanol-sensitive pathways. Transfer of Lucifer yellow between cells also was inhibited by octanol. These observations demonstrate functional gap junctions between cultured alveolar epithelial cells and suggest that gap junctional expression and gap junction-mediated cell-cell coupling are regulated with time in culture.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume272
Issue number6 16-6
StatePublished - Jun 1997
Externally publishedYes

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Keywords

  • Cell-cell communication
  • Communicating junctions
  • Connexins
  • Gene expression
  • Type II cells

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology
  • Physiology (medical)

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