Attenuation of pulmonary neuroendocrine differentiation in mice lacking Clara cell secretory protein.
During development and injury, pulmonary neuroendocrine (NE) cells may transiently express Clara cell 10 kD protein (CC10), a major product of the nonciliated progenitor cells for normal and neoplastic airway epithelia suggesting a close relationship between the cells. To assess the role of CC10 during NE differentiation, we studied CC10-deficient mouse lungs by immunohistochemistry and digital imaging. The knockout model revealed a lack of the disrupted gene product in the lung. Because NE cells, which occur as solitary cells or in neuroepithelial bodies (NEBS), comprise less than 1% of airway epithelia, we counted foci positive for each of the three NE markers--synaptophysin, calcitonin gene-related peptide (CGRP), and protein gene product (PGP) 9.5--and developed a method to analyze numerous airways in serial sections. Digitized images of slides were segmented with Photoshop imaging software. The length of airway epithelium and total section areas were then measured using MetaMorph image analysis software. A comparable range of NE foci was observed regardless of CC10 expression patterns with all three markers, suggesting that CC10 is not critical for NE ontogenesis. However, discrimination according to size revealed that wild-type lungs harbored 30% to 40% greater synaptophysin- and CGRP-containing NEBs relative to CC10 deficient lungs. We posit that an attenuation of pulmonary NE differentiation afflicts the CC10-deficient state. Our imaging application greatly facilitates the acquisition and analysis of complex structures such as the lung and promises to be a widely applicable technique for assessments of tissue sections.
Castro
C M
CM
Medicine Branch, National Cancer Institute, National Institutes of Health, Rockville, Maryland, USA.
Yang
Y
Y
Zhang
Z
Z
Linnoila
R I
RI
D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis; D002454:Cell Differentiation; D004847:Epithelial Cells / Q000502:physiology; D005838:Genotype; D008168:Lung / Q000166:cytology*; D051379:Mice; D018345:Mice, Knockout; D009490:Neurosecretory Systems / Q000166:cytology*; D011506:Proteins / Q000032:analysis / Q000502:physiology*; D014598:Uteroglobin
D011506:Proteins; C556390:Scgb1a1 protein, mouse; D014598:Uteroglobin; D015740:Calcitonin Gene-Related Peptide