Bromodeoxyuridine (BrdU)-label-retaining cells in mouse terminal bronchioles

Histol Histopathol. 2014 May;29(5):659-68. doi: 10.14670/HH-29.10.659. Epub 2013 Dec 4.

Abstract

Adult male mice were continuously treated with bromodeoxyuridine (BrdU) for 1, 2, or 4 weeks by an osmotic pump. To detect BrdU-label-retaining cells (LRCs), putative progenitor/stem cells, other animals were continuously treated with BrdU for 2 weeks, and were then kept without any treatments for 2, 6, or 18 months. The lungs were fixed with 4% paraformaldehyde, and were paraffin-embedded. We observed terminal bronchioles with BrdU immunostaining alone or with BrdU immunostaining accompanying immunostaining for Clara cell secretory protein (CCSP), forkhead box protein J1 (FoxJ1), or calcitonin gene-related peptide (CGRP). The average incidences of BrdU-incorporated cells in the terminal bronchioles after 1, 2, and 4 weeks of continuous BrdU infusion were 6.2%, 11.9%, and 23.1%, respectively. Most BrdU-incorporated cells in these periods were CCSP-immunoreactive (91.7%, 91.3%, and 88.2%, respectively), which means progenitor function of Clara cells. FoxJ1-immunoreactive BrdU-incorporated cells were fewer (5.4%, 3.0%, 2.7%, respectively). The average incidences of BrdU-LRCs in the terminal bronchioles after 2, 6, and 18 months were 7.2%, 4.3, and 2.7%, respectively. Most BrdU-LRCs were CCSP-immunoreactive (91.0%, 92.7%, and 89.6%, respectively), and FoxJ1-immunoreactive BrdU-LRCs were fewer (6.0%, 5.7%, and 2.1%, respectively). CGRP-positive BrdU-incorporated cells were occasional. CGRP-positive BrdU-LRCs were detected in 17.6% of neuroepithelial bodies (NEBs) at 2 months, but disappeared at 6 months. BrdU-positive stem cell candidates, which locate at the brochiolo-alveolar duct junction or cover NEB, were few throughout this study. In conclusion, in the lungs treated only with BrdU, CCSP-immunoreactive cells are important to maintain homeostasis in the terminal bronchiolar epithelium.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells / cytology*
  • Adult Stem Cells / metabolism*
  • Animals
  • Bromodeoxyuridine / administration & dosage
  • Bromodeoxyuridine / pharmacokinetics*
  • Bronchioles / cytology*
  • Bronchioles / metabolism*
  • Calcitonin Gene-Related Peptide / metabolism
  • Forkhead Transcription Factors / metabolism
  • Immunohistochemistry
  • Male
  • Mice
  • Mice, Inbred ICR
  • Neuroepithelial Bodies / cytology
  • Neuroepithelial Bodies / metabolism
  • Pulmonary Alveoli / cytology
  • Pulmonary Alveoli / metabolism
  • Respiratory Mucosa / cytology
  • Respiratory Mucosa / metabolism
  • Time Factors
  • Uteroglobin / metabolism

Substances

  • FOXJ1 protein, mouse
  • Forkhead Transcription Factors
  • Uteroglobin
  • Bromodeoxyuridine
  • Calcitonin Gene-Related Peptide