Quantitative comparison of intracellular concentration and volume of Clara cell 10 KD protein in rat bronchi and bronchioles based on laser scanning confocal microscopy

D. E. Dodge, R. B. Rucker, G. Singh, C. G. Plopper

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

We used an antibody to rat Clara cell 10 KD secretory protein (CC10) to compare the abundance of CC10 in nonciliated epithelium of bronchioles and bronchi by immunohistochemistry and laser scanning confocal microscopy (LSCM) in the reflectance mode. Three zones of reflectance intensity (high, medium, low), directly related to CC10 content, were used to distinguish subcellular compartments of differing densities. Two major compartments contained CC10: granules and endoplasmic reticulum. Bronchiolar cell granules had relatively even reflectance, a high CC10 concentration (0.92 mg/ml), and occupied 7.5% of the cell volume. Bronchial cell granules were bi-zonal, lower in CC10 concentration (0.83 mg/ml), and occupied less cell volume (2.3%). Low- reflecting endoplasmic reticulum occupied a greater cell volume in bronchioles than in bronchi (32.8% and 22.1%, respectively). An intermediate- density zone consisted of endoplasmic reticulum close to granules. CC10 abundance, expressed as ng stored per unit surface area of epithelial basal lamina, was more than three times greater in bronchioles than in proximal bronchi (1.50 ng/mm2 vs 0.42 ng/mm2). These data demonstrate that the process of CC10 secretion differs markedly in non-ciliated epithelium of bronchi and bronchioles. Identifiable mucous-goblet cells did not contain CC10. The LSCM provides a method for quantifying intracellular CC10 pools in intact lung cells and has the potential for quantifying other proteins in subcellular compartments.

Original languageEnglish (US)
Pages (from-to)1171-1183
Number of pages13
JournalJournal of Histochemistry and Cytochemistry
Volume41
Issue number8
DOIs
StatePublished - Jan 1 1993
Externally publishedYes

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Bronchioles
Bronchi
Cell Size
Confocal Microscopy
Endoplasmic Reticulum
Proteins
Epithelium
Goblet Cells
Secretory Pathway
Basement Membrane
Immunohistochemistry
Lung
Antibodies

Keywords

  • Clara cell
  • Clara cell 10 KD protein
  • Laser scanning confocal microscopy
  • Non-ciliated epithelial secretory cell
  • Proximal bronchus
  • Rat lung
  • Serous cell
  • Terminal bronchiole

ASJC Scopus subject areas

  • Anatomy
  • Histology

Cite this

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title = "Quantitative comparison of intracellular concentration and volume of Clara cell 10 KD protein in rat bronchi and bronchioles based on laser scanning confocal microscopy",
abstract = "We used an antibody to rat Clara cell 10 KD secretory protein (CC10) to compare the abundance of CC10 in nonciliated epithelium of bronchioles and bronchi by immunohistochemistry and laser scanning confocal microscopy (LSCM) in the reflectance mode. Three zones of reflectance intensity (high, medium, low), directly related to CC10 content, were used to distinguish subcellular compartments of differing densities. Two major compartments contained CC10: granules and endoplasmic reticulum. Bronchiolar cell granules had relatively even reflectance, a high CC10 concentration (0.92 mg/ml), and occupied 7.5{\%} of the cell volume. Bronchial cell granules were bi-zonal, lower in CC10 concentration (0.83 mg/ml), and occupied less cell volume (2.3{\%}). Low- reflecting endoplasmic reticulum occupied a greater cell volume in bronchioles than in bronchi (32.8{\%} and 22.1{\%}, respectively). An intermediate- density zone consisted of endoplasmic reticulum close to granules. CC10 abundance, expressed as ng stored per unit surface area of epithelial basal lamina, was more than three times greater in bronchioles than in proximal bronchi (1.50 ng/mm2 vs 0.42 ng/mm2). These data demonstrate that the process of CC10 secretion differs markedly in non-ciliated epithelium of bronchi and bronchioles. Identifiable mucous-goblet cells did not contain CC10. The LSCM provides a method for quantifying intracellular CC10 pools in intact lung cells and has the potential for quantifying other proteins in subcellular compartments.",
keywords = "Clara cell, Clara cell 10 KD protein, Laser scanning confocal microscopy, Non-ciliated epithelial secretory cell, Proximal bronchus, Rat lung, Serous cell, Terminal bronchiole",
author = "Dodge, {D. E.} and Rucker, {R. B.} and G. Singh and Plopper, {C. G.}",
year = "1993",
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TY - JOUR

T1 - Quantitative comparison of intracellular concentration and volume of Clara cell 10 KD protein in rat bronchi and bronchioles based on laser scanning confocal microscopy

AU - Dodge, D. E.

AU - Rucker, R. B.

AU - Singh, G.

AU - Plopper, C. G.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - We used an antibody to rat Clara cell 10 KD secretory protein (CC10) to compare the abundance of CC10 in nonciliated epithelium of bronchioles and bronchi by immunohistochemistry and laser scanning confocal microscopy (LSCM) in the reflectance mode. Three zones of reflectance intensity (high, medium, low), directly related to CC10 content, were used to distinguish subcellular compartments of differing densities. Two major compartments contained CC10: granules and endoplasmic reticulum. Bronchiolar cell granules had relatively even reflectance, a high CC10 concentration (0.92 mg/ml), and occupied 7.5% of the cell volume. Bronchial cell granules were bi-zonal, lower in CC10 concentration (0.83 mg/ml), and occupied less cell volume (2.3%). Low- reflecting endoplasmic reticulum occupied a greater cell volume in bronchioles than in bronchi (32.8% and 22.1%, respectively). An intermediate- density zone consisted of endoplasmic reticulum close to granules. CC10 abundance, expressed as ng stored per unit surface area of epithelial basal lamina, was more than three times greater in bronchioles than in proximal bronchi (1.50 ng/mm2 vs 0.42 ng/mm2). These data demonstrate that the process of CC10 secretion differs markedly in non-ciliated epithelium of bronchi and bronchioles. Identifiable mucous-goblet cells did not contain CC10. The LSCM provides a method for quantifying intracellular CC10 pools in intact lung cells and has the potential for quantifying other proteins in subcellular compartments.

AB - We used an antibody to rat Clara cell 10 KD secretory protein (CC10) to compare the abundance of CC10 in nonciliated epithelium of bronchioles and bronchi by immunohistochemistry and laser scanning confocal microscopy (LSCM) in the reflectance mode. Three zones of reflectance intensity (high, medium, low), directly related to CC10 content, were used to distinguish subcellular compartments of differing densities. Two major compartments contained CC10: granules and endoplasmic reticulum. Bronchiolar cell granules had relatively even reflectance, a high CC10 concentration (0.92 mg/ml), and occupied 7.5% of the cell volume. Bronchial cell granules were bi-zonal, lower in CC10 concentration (0.83 mg/ml), and occupied less cell volume (2.3%). Low- reflecting endoplasmic reticulum occupied a greater cell volume in bronchioles than in bronchi (32.8% and 22.1%, respectively). An intermediate- density zone consisted of endoplasmic reticulum close to granules. CC10 abundance, expressed as ng stored per unit surface area of epithelial basal lamina, was more than three times greater in bronchioles than in proximal bronchi (1.50 ng/mm2 vs 0.42 ng/mm2). These data demonstrate that the process of CC10 secretion differs markedly in non-ciliated epithelium of bronchi and bronchioles. Identifiable mucous-goblet cells did not contain CC10. The LSCM provides a method for quantifying intracellular CC10 pools in intact lung cells and has the potential for quantifying other proteins in subcellular compartments.

KW - Clara cell

KW - Clara cell 10 KD protein

KW - Laser scanning confocal microscopy

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KW - Serous cell

KW - Terminal bronchiole

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