Heterogeneity of Clara cell glutathione

A possible basis for differences in cellular responses to pulmonary cytotoxicants

Jay A A West, Christine H. Chichester, Alan R Buckpitt, Nancy K. Tyler, Patricia Brennan, Cimity Helton, Charles Plopper

Research output: Contribution to journalArticle

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Abstract

Clara-cell populations show a high degree of variation in susceptibility to injury by bioactivated cytotoxicants. Because glutathione (GSH) is critical for detoxification of electrophilic metabolites, heterogeneity in Clara cell GSH levels may lead to a wide range of cytotoxic responses. This study was designed to define the distinct GSH pools within Clara cells, characterize heterogeneity within the population, and examine whether heterogeneity contributes to susceptibility. Using fluorescent imaging combined with high-performance liquid chromatography analysis, semiquantitative measurements were obtained by evaluation of GSH using monochlorobimane and monobromobimane. In steady-state conditions, the GSH measured in isolated cells was in the femtomole range, but varied 4-fold between individual cells. Clara cells analyzed in situ and in vitro confirmed this heterogeneity. The response of these cells to compounds that modulate GSH was also variable. Diethylmaleate depleted GSH, whereas GSH monoethylester augmented it. However, both acted nonuniformly in isolated Clara cells. The depletion of intracellular GSH caused a striking decrease in cell viability upon incubation with naphthalene (NA). The sulfhydryl-binding fluorochrome BODIPY, which colocalized with tetramethylrosamine, a mitochondrial dye, demonstrated by confocal microscopy that cellular sulfhydryls are highest in the mitochondria, next-highest in cytoplasm, and lowest in the nucleus. These pools responded differently to modulators of GSH. We concluded that the steady-state intracellular GSH of Clara cells exists in distinct pools and is highly heterogeneous within the population, and that the heterogeneity of GSH levels corresponds closely to the response of Clara cells to injury by NA.

Original languageEnglish (US)
Pages (from-to)27-36
Number of pages10
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume23
Issue number1
StatePublished - 2000

Fingerprint

diethyl maleate
Glutathione
Cells
Lung
Detoxification
Mitochondria
Confocal microscopy
High performance liquid chromatography
Metabolites
Fluorescent Dyes
Modulators
Coloring Agents
Imaging techniques
Population Characteristics
naphthalene
Wounds and Injuries
Confocal Microscopy
Cell Survival
Cytoplasm
High Pressure Liquid Chromatography

ASJC Scopus subject areas

  • Cell Biology
  • Pulmonary and Respiratory Medicine
  • Molecular Biology

Cite this

Heterogeneity of Clara cell glutathione : A possible basis for differences in cellular responses to pulmonary cytotoxicants. / West, Jay A A; Chichester, Christine H.; Buckpitt, Alan R; Tyler, Nancy K.; Brennan, Patricia; Helton, Cimity; Plopper, Charles.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 23, No. 1, 2000, p. 27-36.

Research output: Contribution to journalArticle

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