M. H. A. De Groodt-Lasseel

Nitric oxide synthase immunoreactivity in the enteric nervous system of the developing human digestive tract.

We have investigated indirectly the presence of nitric oxide in the enteric nervous system of the digestive tract of human fetuses and newborns by nitric oxide synthase (NOS) immunocytochemistry and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) histochemistry. In the stomach, NOS immunoactivity was confined to the myenteric plexus and nerve fibres in the outer smooth musculature; few immunoreactive nerve cell bodies were found in ganglia of the outer submucous plexus. In the pyloric region, a few nitrergic perikarya were seen in the inner submucous plexus and some immunoreactive fibres were found in the muscularis mucosae. In the small intestine, nitrergic neurons clustered just underneath or above the topographical plane formed by the primary nerve strands of the myenteric plexus up to the 26th week of gestation, after which stage, they occurred throughout the ganglia. Many of their processes contributed to the dense fine-meshed tertiary nerve network of the myenteric plexus and the circular smooth muscle layer. NOS-immunoreactive fibres directed to the circular smooth muscle layer originated from a few NOS-containing perikarya located in the outer submucous plexus. In the colon, caecum and rectum, labelled nerve cells and fibres were numerous in the myenteric plexus; they were also found in the outer submucous plexus. The circular muscle layer had a much denser NOS-immunoreactive innervation than the longitudinally oriented taenia. The marked morphological differences observed between nitrergic neurons within the developing human gastrointestinal tract, together with the typical innervation pattern in the ganglionic and aganglionic nerve networks, support the existenc of distinct subpopulations of NOS-containing enterice neurons acting as interneurons or (inhibitory) motor neurons.

Myocardial ultrastructure and haemodynamic reactions during experimental subarachnoid haemorrhage

Abstract The mechanisms governing the haemodynamic, electrocardiographic and ultrastructural changes following subarachnoid haemorrhage are studied. The experiments on dogs show that an increased cellular metabolism due to noradrenaline and to adrenaline, is an essential factor in the haemodynamic reactions in subarachnoid haemorrhage. During the stimulation stage of these hormones, ultrastructural changes appear, indicating an important migration of calcium ions from their store to their point of action. In so far as they influence the action potentials, these movements initially create a “hyperkalaemic” type ECG probably in relation with an associated potassium ion displacement. In the second stage, an “ischaemic” type of ECG change is found which is totally independent of hypoxia, but also caused by the increase in cellular metabolism. This second stage is still reversible. The last or third stage occurs when the hormones deeply disturb the cellular metabolism because of their high dose or because of their prolonged action. At this point irreversible and destructive cellular alterations occur.

Pulmonary macrophages in birds (barn owl, Tyto tyto alba), domestic fowl (Gallus gallus f. domestica), Quail (Coturnix coturnix), and pigeons (Columbia livia)

Birds have a limited number of resident macrophages in the normal steady‐state respiratory tract. The discovery of phagocytes in lavages of lung from birds contrasts with findings that phagocytes are seldom seen in investigations in situ. An electron microscopic study was performed in the respiratory units, the parabronchi, and air capillaries in particular in several adult bird species to localize the seat of respiratory macrophages.

Calcitonin Gene-Related Peptide-Like Immunoreactivity in the Human Small Intestine

Calcitonin-gene-related-peptide (CGRP)-like immunoreactivity was localized in nerve fibres, neuronal somata and in mucosal endocrine cells of the human small intestine. Immunoreactive enteric neurons were more numerous in the submucous plexuses than in the myenteric plexus. Morphologically, they predominantly had the appearance of type II neurons. The majority of the CGRP-like immunoreactive nerve fibres ran within the ganglionic nerve plexuses. Only a small proportion could be observed in the lamina propria, the lamina muscularis mucosae, or the circular and longitudinal outer smooth muscle layer. These findings suggest that within the wall of the human small intestine neuronal CGRP of either extrinsic or intrinsic origin exerts its effect chiefly on other enteric neurons, and might be indirectly involved in the regulatory functions of the human small intestine.

A correlative light-, fluorescence- and electron-microscopic study of neuroepithelial bodies in the lung of the red-eared turtle, Pseudemys scripta elegans

SummaryNeuroepithelial bodies (NEB) were identified for the first time in the respiratory tract of a reptile by the use of combined morphological and histochemical methods. In the red-eared turtle, Pseudemys scripta elegans, NEB were found within the trabecular epithelium of the respiratory tract, mainly in the branching regions of the trabeculae. An intracellular formaldehyde-induced fluorescent compound was identified as 5-hydroxytryptamine (5-HT) by means of microspectrofluorometry. Subsequent histochemical staining of the same fluorescent sections showed the 5-HT-containing cells to be argentaffin. In electron micrographs cell clusters characterized by the presence of distinctive cytoplasmic, membrane-bounded dense granules (±100 nm) were observed, correlating with the distribution of the yellow-fluorescent epithelial cells. The granules of the NEB are positive when the argentaffin technique is performed directly on ultrathin sections. Cells of the NEB extend into the lumen of the airway via apical microvilli and a single modified cilium displaying a 9×2+0 or 8×2+2 microtubular pattern. Unmyelinated axons containing mostly small, clear vesicles were seen in close association with NEB cells, often forming synaptic junctions. Occasionally, axons containing a few small dense-cored vesicles were found. The relationship between NEB cells and capillaries, the images of emiocytotic granule release and the occurrence of synaptic contacts between axons and granule-containing cells are indicative of endocrine secretion. These features and the presence of intracytoplasmic granules containing 5-HT may justify the inclusion of NEB-cells of the turtle lung into the diffuse neuroendocrine system. Furthermore, structurally these cells appear to represent sensory elements capable of an intrapulmonary recepto-secretory function.

Occurrence, distribution and neurochemical features of small intestinal neurons projecting to the cranial mesenteric ganglion in the pig

The small intestine of the pig has been investigated for its topographical distribution of enteric neurons projecting to the cranial mesenteric ganglion, by using Fast Blue or Fluorogold as a retrogradely transported neuronal tracer. Contrary to the situation in small laboratory animals such as rat and guinea-pig, the intestinofugally projecting neurons in the porcine small intestine were not restricted to the myenteric plexus, but were observed in greater numbers in ganglia of the outer submucous plexus. The inner submucous plexus was devoid of labelled neurons. Retrogradely labelled neurons were mostly found, either singly or in small aggregates, in ganglia located within a narrow border on either side of the mesenteric attachment. For both nerve networks, their number increased from duodenum to ileum. All the retrogradely labelled neurons exhibited a multidendritic uniaxonal appearance. Some of them displayed type-III morphology and stained for serotonin. This study indicates that, in the pig, not only the myenteric plexus but also one submucous nerve network is involved in the afferent component of intestino-sympathico-intestinal reflex pathways. The finding that some of the morphologically defined type-III neurons participate in these reflexes is in accord with the earlier proposal that type-III neurons are supposed to fulfill an interneuronal role, whether intra- or extramurally.

AN ELECTRON MICROSCOPIC STUDY OF THE PARABRONCHIAL EPITHELIUM IN THE MATURE LUNG OF FOUR BIRD SPECIES

No integrated comprehensive description of the ultrastructure of the parabronchial epithelium is available. The origin, discharge, and occurrence of the trilaminar substance have not yet been sufficiently studied. Therefore, the main objectives were to classify the cell types of the parabronchial epithelium and to describe their role in manufacturing the trilaminar substance.

Immunohistochemical localization of calbindin-D28K in the brain of a cartilaginous fish, the dogfish (Scyliorhinus canicula L.).

The occurrence and distribution of the vitamin-D-induced calcium-binding protein, calbindin-D28K, has been studied in the brain of a cartilaginous fish using immunohistochemical techniques. A strong immunoreactivity was found in the perikarya, dendrites and axons of neurons located in the nucleus interstitialis commissurae anterioris, the nucleus medialis of the left habenula, the thalamus dorsalis, the thalamus ventralis, the nucleus lobi lateralis, the nucleus interpeduncularis, the lobus vagi and the medial reticular zone. Fibre tracts associated with some of these neuronal groups, such as the fasciculus retroflexus, the stria medullaris and the commissura habenulae, also contained immunopositive fibres. Only a minor immunoreactivity could be detected in other brain areas such as the tectum mesencephali and some telencephalic zones. Interestingly, the cerebellum did not show any immunoreactivity in Purkinje cells nor in other neurons. The distribution of calbindin-D28K in the dogfish brain appears to be mainly related to the viscerosensory centres.

Anchoring and support system of pulmonary gas-exchange tissue in four bird species.

Avian air capillaries are delicate structures compared to the mammalian pulmonary alveolus. A transmission and scanning electron microscopic study was carried out on several species of birds with the aim of determining the support structures of the avian gas-exchange mantle. Lung tissue of two bird species belonging to strong flying birds (pigeon and barn owl) and two relatively flightless species (domestic fowl and quail) was subjected to standard processing for transmission and scanning electron microscopy after intratracheal inflation. Twisted profiles of lipoproteinaceous trilaminar substance as specific secretory product of avian squamous respiratory cells can be seen in the cell body and cytoplasmic extensions that are wedged between the blood capillaries, partly surrounding them. The intracytoplasmatically located trilaminar complexes form a three-dimensional intricate spiderweb-like system between the blood capillaries and air capillaries, which presumably function as an anchoring and support structure of the gas-exchange tissue. This system is strengthened by retinacula--pairs of attenuated parallel processes of squamous respiratory cells that project to the airway lumen--expanding and bridging the opposite side of air capillaries. The trilaminar substance is discharged in the form of a 15-nm-thick acellular lining layer which is uniquely adapted to the extremely thin respiratory epithelium. The trilaminar substance arises in the cytoplasm of squamous respiratory cells from profiles of granular and smooth endoplasmic reticulum. The integrity and stability of the gas-exchange tissue is likely to be guaranteed by a specific arrangement of the squamous respiratory cells, in which the trilaminar substance plays a paramount role. This general pattern can be observed in strong flying bird species as in the relatively flightless birds.

Neuroepithelial endocrine cells in the lung of the lungfish Protopterus aethiopicus : an electron- and fluorescence-microscopical investigation

The occurrence and distribution of neuroepithelial endocrine (NEE) cells was demonstrated electron- and fluorescence-microscopically in the lungfish Protopterus aethiopicus. They were only found to occur solitarily in the basal part of the cilio-mucous epithelium which is restricted to the pneumatic duct and adjacent parts of the common anterior chamber. The NEE cells show a yellow, formaldehyde-induced fluorescence. Electron-microscopically, all the NEE cells are characterized by membrane-bound electron-dense secretory granules with varying diameters, ranging from 75 to 150 nm. These granules are distributed throughout the cytoplasm with a higher concentration in the basal region. The NEE cells were regularly found to contain paracrystalline inclusions with a tubule-like substructural arrangement. A small part of the NEE cells appeared to reach the luminal surface by means of a long slender process bearing specialized beaded microvilli on its apical pole. Intraepithelial nerve fibres, with the ultrastructural characteristics of afferent fibres, were found running parallel to the airway surface. Nerve profiles, largely resembling the latter, can be seen in the proximity of the basolateral plasma membrane of the NEE cells. In addition, nerve terminals containing an aggregation of small clear vesicles are in close contact with the NEE cells. In conclusion, it appears that, as has so far been assumed in higher vertebrates, the NEE cells in the lung of Protopterus may perceive changes in the airway gases whereupon they could respond by releasing a chemical modulator, influencing contacting afferent nerve terminals or nearby smooth muscle bundles. Furthermore, intraepithelial nerve fibres or NEE cells might be stretch-sensitive.