Exocytosis in alveolar type II cells revealed by cell capacitance and fluorescence measurements

Norbert Mair, Thomas Haller, Paul Dietl, P. W. Shaul

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Measurement of lamellar body (LB) exocytosis at high spatial and temporal resolution was recently enabled by fluorescence of the dye FM 1-43 (F(FM1-43)). Here, the capabilities of this method were further examined and extended by simultaneous measurement of the cell membrane capacitance (C(m)) and laser-scanning confocal microscopy. Step increases in C(m) were evoked by extracellular ATP (20 μM) or an elevated pipette Ca2+ concentration (≥3 μM). The delay between the first C(m) step and the increase in F(FM1-43) was <1 s, indicating ready access of FM 1-43 to exocytosed LB contents. A specific C(m) of 0.88 μF/cm2 for the membrane of an exocytosed LB was calculated. Compound exocytosis was occasionally observed. Decreases in C(m), indicative of transient fusion or endocytosis, did not occur within 20 min of stimulation. Exocytosis was stimulated by 160 μM guanosine 5'-O-(3- thiotriphosphate) in the pipette, but compound exocytosis was unaffected. The comparison of methods revealed that FM 1-43 is ideally suited to measure the onset of exocytosis and amount of secretion. Patch clamp is superior in resolving fusion events with the plasma membrane.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume276
Issue number2 20-2
StatePublished - Feb 1999

Fingerprint

Alveolar Epithelial Cells
Exocytosis
Fluorescence
Cell Membrane
Coloring Agents
Guanosine 5'-O-(3-Thiotriphosphate)
Endocytosis
Confocal Microscopy
Adenosine Triphosphate
Membranes
FM1 43

Keywords

  • Compound exocytosis
  • Endocytosis
  • Patch clamp
  • Surfactant

ASJC Scopus subject areas

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

Cite this

Exocytosis in alveolar type II cells revealed by cell capacitance and fluorescence measurements. / Mair, Norbert; Haller, Thomas; Dietl, Paul; Shaul, P. W.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 276, No. 2 20-2, 02.1999.

Research output: Contribution to journalArticle

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