Ontogeny of neuroendocrine cells in human fetal lung. III. An electron microscopic immunohistochemical study

Immunocytochemistry at the transmission electron microscopic level utilizing colloidal gold spheres conjugated with secondary antisera was performed on lungs of 22 human fetuses and newborn infants of 13 to 38 weeks gestation and from birth to 5 1/2 months of postnatal life. Tissue was stained for the peptide hormones, immunoreactive (IR) bombesin, and IR calcitonin. In addition to unmatched neuroendocrine (NE) cells identified for these peptides, matched cells were identified in near-serial ribbons, each stained for an individual peptide. Based on morphology, five subtypes of NE cells were examined for these two peptides. We have confirmed the previously demonstrated developmental appearances of these peptides in human fetal lung. We also have found many cells containing both peptides in the lungs of live-born infants of 25 weeks gestation or more who survived long enough to develop chronic lung disease. The percentage of neurosecretory granules labeled for IR bombesin which overlapped with the percentage of granules labeled for IR calcitonin in cells of several dysplastic lung suggested that both peptides could be contained within a single granule. This was confirmed in NE cells of four such infants in preliminary studies utilizing double labeling immunocytochemistry at the electron microscopic level. At least two subpopulations of NE cells were not labeled for either peptide, suggesting that as yet-unidentified peptides and/or amines are contained in their granules. The possibility that large nonlabeled granules contain hormone precursors is also raised.