Immunocytochemical localization of aromatic L-amino acid decarboxylase in human, rat, and mouse bronchopulmonary and gastrointestinal endocrine cells

Aromatic L-amino acid decarboxylase (AADC) catalyzes the cellular decarboxylation of L-aromatic amino acids and is therefore involved in the synthesis of several biogenic amines. Application of the indirect immunoperoxidase method on human, rat, and mouse tissues using specific antibodies to AADC revealed all AADC-containing cells. Besides mast cells and adrenergic nerve fibers, the following cells were immunostained: neuroendocrine cells in the tracheobronchial epithelium; neuroepithelial bodies in the bronchopulmonary epithelium; Kultschitzky cells in the small intestine and appendix as well as adrenal chromaffin cells. All the latter cells belong to the so-called APUD system, the "D" in the acronym standing for the activity of the enzyme aromatic L-amino acid decarboxylase. Immunocytochemistry for AADC may become an additional tool not only to highlight APUD cells in tissue sections but also to differentiate the sites of cellular amine synthesis from those of amine storage.     

Sarafotoxin expression in the bronchopulmonary tract: immunohistochemical occurrence and colocalization with endothelins

The immunohistochemical occurrence of sarafotoxin (SRTX), a snake venom peptide under strong evolutionary control, was investigated in the pulmonary diffuse neuroendocrine system (PDNES) of newborn cats and rats. By applying the avidin-biotin-peroxidase complex method on serial lung sections, we have demonstrated its distribution and colocalization with different endothelin (ET) isoforms. A light microscopic study revealed apparent immunostaining for SRTX in neuronal components and smooth muscle tissue and in neuroepithelial bodies (NEB), while isolated neuroendocrine cells (NEC) remain unlabelled. Comparison of the SRTX reactivity pattern with that of different ET peptides on adjacent lung sections showed colocalization of SRTX-b with ET-3 in NEB, intrapulmonary ganglion cells and nerve fibres, on the one hand, and with ET-1 in airway and vascular smooth muscle cells, on the other. These findings, in addition to the remarkable functional and structural similarities between SRTX and ET peptides, suggest a common evolutionary origin and biological significance of sarafotoxin and endothelins. Moreover, this is the first time that a toxic peptide has been demonstrated in the PDNES.     

Immunocytochemical Localization of Serotonin and Neuropeptides in the Neuroendocrine Paraneurons of Teleost and Lungfish Gills*

Neuroendocrine (NE) cells in the gills of six fish species were identified by serotonin and peptide immunohistochemistry. They occur either as single cells or as cell clusters within the filamental epithelium. Two populations of NE cells were identified that exhibit an immunoreactivity either for serotonin alone or for met-enkephalin and leu-enkephalin co-distributed in the same cells. We demonstrate for the first time the occurrence of open-type endocrine cells in the fish gill epithelium and confirm the presence of only serotonin-immunopositive cells of the closed type. The NE cells of the species studied do not react positively to other neuroendocrine markers occurring in the mammalian lung. Great differences probably exist in the expression of these epitopes on antigenic structures among various animal species. Although the physiological role of neuropeptides and serotonin in the fish gill NE cells is unknown, the presence of these molecular markers is of interest not only from an evolutionary aspect, but also in terms of a potential animal model to study the supposed functions of the airway neuroendocrine system in the vertebrates.     

The Meckel syndrome. Pathological and cytogenetic observations in eight cases

Summary Eight new cases of Meckel syndrome, two of them occurring in the same family, are presented. Occipital encephalocele of varying extent, multicystic renal dysplasia not associated with urinary tract obstruction, and postaxial hexadactyly comprise the three basic features of this lethal syndrome with autosomal recessive inheritance. From our observations it appears that congenital hepatic fibrosis, abnormal external genitalia in male infants and a malformed tongue with lipomatous excrescences are also frequently occurring anomalies with important diagnostic value.The statement that the majority of cases of Meckel syndrome can be detected prenatally is further supported by two cases in the present series. The incidence of this syndrome may be much higher than previously thought.     

Microspectrographic analysis of formaldehyde-induced fluorescence in the quail adenohypophysis after injection of l-dopa and 5-hydroxytryptophan

Summary The spectral distribution of the formaldehyde-induced fluorescence emitted by model solutions and by adenohypophyses after intraperitoneal injection of l-dopa or 5-hydroxytryptophan was analyzed microspectrographically. Based on previously reported studies and on present findings, it seems that dopamine is stored in the strongly fluorescent cells after injection of l-dopa, and that a compound closely related to 5-hydroxytryptophan or serotonin is present in most of the cells after injection of 5-hydroxytryptophan. A non-specific, granular fluorescence appeared after 5-hydroxytryptophan and, to a lesser extend, l-dopa treatment. It probably represents autofluorescence of lysosomes, which are numerous in these circumstances.     

Fine structure and serotonin immunohistochemistry of the neuroendocrine cells in the lungs of the bichirs Polypterus delhezi and P. ornatipinnis

Neuroendocrine cells (NE) occurring in the pulmonary epithelium of the fishes Polypterus delhezi and P. ornatipinnis are studied by electron microscopy and by immunostaining for serotonin which is often present in such cells in the mammalian lung. With the electron microscopy NE are found to occur single, resting upon the basement membrane and forming a narrow cytoplasmic extension towards the air lumen. They contain dense-cored vesicles of 80-165 nm which form exocytotic profiles at the level of the basal membrane. An immunoreactivity for serotonin is demonstrated for the first time in the NE of these species. The role of this mediator may involve a paracrine or endocrine function as postulated for the respiratory neuroendocrine mammalian cells. NE of the species studied are considered similar to those found within the wall of lung airways in mammals and submammalian vertebrates. Although much immunocytochemical investigations remain to be executed, they may also be included in the APUD (or DNES) cell system.     

The ultrastructure of lymphatic valves in the adult rabbit lung

Summary An electron microscopic investigation has revealed that the pulmonary lymphatic valves of adult rabbits are not simple duplicatures of the lymphatic vessel wall. They consist of an uninterrupted central connective tissue core, covered on both sides with a single layer of flattened endothelial cells. Near their insertion in the lymphatic vessel wall, the connective tissue core reveals a distinct thickening being composed mainly of collagen bundles. In the other parts it contains mainly elastic fibers and fine filaments, enclosing also some rather peculiar connective tissue cells. Nervous and muscular elements were not observed. The endothelium is continuous and exhibits no open junctions. The valvular basement membrane is better developed than in lymphatic capillaries. The endothelial cells contain numerous cytoplasmic filaments which might be endowed with contractile properties. The nuclei of the endothelial and the connective tissue cells are irregularly spaced and frequently clustered near the free edge of the valve.These ultrastructural features suggest that the function of the lymphatic valves is mainly passive. They are firmly inserted in the lymphatic vessel wall by collagen fibers and their moving parts are slender and elastic. Their endothelium appears relatively impermeable and is firmly attached to the subjacent connective tissue.This study has been supported by a grant from The Council for Tobacco Research—U.S.A.. We thank Professor Robert C. Rosan (Saint Louis University—U.S.A.) for expert advice, R. Janssens for technical, G. Pison and St. Ons for photographic and N. Tyberghien for secretarial assistance.