Am. J. Respir. Cell Mol. Biol.,
Volume 19, Number 2, August, 1998 278-285
Calcitonin Gene-related Peptide Expression Is Altered in Pulmonary
Neuroendocrine Cells in Developing Lungs of Rats with Congenital
Diaphragmatic Hernia
Hanneke
IJsselstijn,
Noelyn
Hung,
Johan C.
de Jongste,
Dick
Tibboel,
and
Ernest
Cutz
Departments of Pediatric Surgery and Pediatrics, Erasmus University and University Hospital/Sophia Children's Hospital,
Rotterdam, The Netherlands; and Department of Pathology and The Research Institute, The Hospital for Sick Children,
Toronto, Ontario, Canada
Congenital diaphragmatic hernia (CDH) is associated with high neonatal mortality from lung hypoplasia
and persistent pulmonary hypertension. Pulmonary neuroendocrine cells (PNEC) produce calcitonin gene-related peptide (CGRP), a potent vasodilator. We previously reported altered distribution of CGRP-positive PNEC in full-term rats with CDH, that may lead to an imbalance in vasoactive mediators. In the
present study we examined the expression of CGRP-positive PNEC during lung development in rats with
CDH induced by 2,4-dichlorophenyl-p-nitrophenylether (Nitrofen). Cesarean sections were performed on
Days 16, 18, 20, or 22, and the lungs were immunostained for CGRP and immunoreactive cells were quantitated through image analysis. On Day 16, CGRP-immunoreactive staining was negative; on Day 18, CGRP-immunoreactive cells were found in all controls (not exposed to Nitrofen), whereas in CDH pups,
CGRP-positive cells were present in only four of six cases. On Day 20, CGRP immunoreactivity was similar in CDH pups, Nitrofen-exposed pups without CDH, and controls. On Day 22 (term), significantly more CGRP-positive cells (i.e., number of positive cells per surface area [mm2] or lung volume [mm3])
were found in ipsilateral lungs of CDH pups than in controls (P < 0.05). The difference was even more
striking in contralateral lungs of CDH pups (P < 0.001), ruling out nonspecific effects of Nitrofen. In
CDH lungs, the proportion of immunostained epithelium and the size of the neuroendocrine cell clusters
(neuroepithelial bodies [NEB]) were not significantly different from those of controls. On Day 22, supraoptimal dilution immunocytochemistry yielded similar results in CDH pups and controls. We conclude
that in CDH, CGRP expression in PNEC and NEB is delayed during early stages of lung development. Because CGRP also exhibits growth factor-like properties for endothelium and epithelial cells, the lack of
this factor during a crucial developmental stage (canalicular period) may be causally related to lung hypoplasia.
