Omeprazole, a substituted benzimidazole, is known to inhibit acid secretion from parietal cells in gastric glands, and is widely utilized as a drug for peptic ulcer. To clarify the ultrastructural changes in parietal cells from long-term treatment with a therapeutic dose of omeprazole, biopsy specimens of the gastric mucosa obtained from peptic ulcer patients were morphometrically analyzed before and after omeprazole treatment. Before treatment with omeprazole, parietal cells in both the stimulated and resting stages were observed; the stimulated cells possessed smaller amounts of tubulovesicles in the cytoplasm and numerous profiles of microvilli in the intracellular canaliculi, whereas the cells in the resting phase showed numerous profiles of tubulovesicles and poorly developed microvilli in the canaliculi. Eight weeks after the onset of omeprazole treatment, the amounts of both tubulovesicles in the cytoplasm and microvilli in the intracellular canaliculi drastically decreased. These decreases in the profiles of the membrane structures with a proton pump occurred concomitantly with a significant increase in autophagic vacuole/autolysosome-like structures. These results suggest that the membrane structures with proton pump are not recycled between tubulovesicles and microvilli of intracellular canaliculi in parietal cells after omeprazole treatment, but may be sequestrated into autophagosomes and degraded by lysosomal enzymes.

View full abstract

The enzyme complex 3β-hydroxysteroid dehydrogenase (3β-HSD) is involved in the biosynthesis of all classes of active steroids. It is known that the enzymatic activity of 3β-HSD is present not only in classical steroidogenic tissues, but also in many peripheral tissues including cardiac tissue. To determine whether 3β-HSD is present in rat non-steroidogenic tissues, we examined cardiovascular tissues including the ventricle, atrium, aortic arch, abdominal aorta, and inferior vena cava by immunohistochemistry and Western blotting using polyclonal antibody raised against a synthetic peptide of human placental 3β-HSD.
By Western blotting, protein bands immunoreactive for anti-3β-HSD were detected at molecular weights of 42 and 37kDa in both the ventricle and atrium, whereas only a 37kDa band was recognized in both the aortic arch and abdominal aorta. By immunohistochemistry, immunoreactivity for 3β-HSD was detected in both the ventricular and atrial cardiocytes, while immunostaining was also found, though faintly, in the smooth muscles of the aortic arch, abdominal aorta, and inferior vena cava.
These results suggest that cardiocytes may synthesize the steroidogenic 3β-HSD enzyme.

View full abstract

Melanin pigments in lower vertebrates are often found in locations other than the skin, thus forming an extracutaneous pigmentary system of unknown function. The cellular and biochemical structure of this system is still poorly characterized. This paper deals with the ultrastructural and biochemical features of the melanogenic system of Xenopus laevis. Melanin containing cells were identified in the dorsal and ventral skin, and in the lung, spleen, liver and connective tissue surrounding blood vessels. The pigment cells in the skin and the lungs appeared to be typical melanocytes. The spleen contained isolated melanocyte-like cells, but most of the pigment cells present in this organ were associated with melanomacrophage centers. Conversely, the liver appeared devoid of melanocytes and only displayed melanomacrophage centers. Tyrosinase activity was found in all pigment-containing organs except the liver. All organs containing tyrosinase activity also displayed melanin formation potential from L-tyrosine. Therefore, tyrosine hydroxylase and melanin formation activities could be detected only in those organs containing typical melanocytes but not in locations such as the liver, where only melanomacrophages centers were found.

View full abstract

An isomer of ouabain, the ouabain-like compound (OLC), may participate in the regulation of body fluid volume and vascular tone. Forebrain regions, especially the hypothalamus, are reported to be sites of OLC action in the central nervous system. The medulla oblongata is another critical area involved in central cardiovascular regulation. We reported that the microinjection of either monoclonal antibody to ouabain T8B11 or Fab fragment of digoxin-specific antibody into the rostral ventrolateral medulla significantly decreased mean arterial pressure and renal sympathetic nerve activity in anesthetized normotensive rats (TERUYA et al.: J. Clin. Invest. 99: 2791-2798, 1997). Using T8B11, we examined the ouabain-like immunoreactivity in the medulla oblongata of normotensive rats. In periodate-lysine-paraformaldehyde fixed tissues, ouabain-like immunoreactive neurons were detected in the nuclei and regions in the medulla oblongata including the ventrolateral medulla, ventromedial medulla, nucleus ambiguus, caudal raphe nuclei, nucleus of solitary tract, and dorsal motor nucleus of the vagus. When an Fab fragment of digoxin-specific antibody was used as a first antibody, the digoxin-like immunoreactive neurons were distributed in almost the same pattern as those observed with the use of T8B11. In the brain fixed with the “three-step” procedure developed by YAMADA et al. (1987), which was used in a previous ouabain immunohistochemical study of the hypothalamus, ouabain-like immunoreactivity in the medulla oblongata was much weaker in intensity and less restricted in distribution than that in the hypothalamus. These findings suggest that ouabain-like immunoreactivities are present in the medulla oblongata with a manner of distribution different from that seen in the hypothalamus. Some ouabain-immunopositive nuclei and regions in the medulla oblongata, especially the rostral ventrolateral medulla, may be other OLC action sites.

View full abstract

The taste buds in the soft palate and the circumvallate papillae of the rat were investigated by the zinc iodide-osmium tetroxide technique. In addition, electron micrographs of taste buds stained with this method were presented for the first time. Differences in taste bud structures were found between the examined regions. The taste buds of the soft palate showed a complicated plexus of intragemmal nerve fibers. Some fibers exhibited terminal polymorphic swellings. Single branches could be traced close to the space of the taste pore. In the soft palate, the taste bud cells remained unstained, whereas in the circumvallate papillae of the tongue, a subpopulation of taste bud cells could be selectively stained and the intragemmal nerve fibers were characterized by large varicosities. The morphological dissimilarities between the taste buds of the investigated regions might be explained by their functional characteristics, or possibly their varying affinities to the taste qualities. Electron microscopic investigation of the stained circumvallate papillae revealed that the electron-dense reaction product had primarily accumulated in a subpopulation of light cells. Dark cells exhibited only a slight labelling. In detail, the precipitate was found loosely distributed in the cytoplasm as well as the nuclei of the cells, and particularly concentrated at the membranes of light vacuoles, this probably being profiles of dilated endoplasmic reticulum. A few roundish accumulations of precipitate were seen in the cytoplasm of taste bud cells, which showed no intensive light microscopic staining. Labelled material was also found within the taste pores outside the apical processes of the cells. The present findings indicate that the zinc iodide-osmium tetroxide technique is applicable to neuroanatomical studies of taste buds.

View full abstract

Chinese hamster metaphase chromosomes were stained with platinum (Pt) blue and observed in the hydrous state with low vacuum scanning electron microscopes (LVSEM). The coiled structure of the chromatin fibers in the chromosomes was well recognized through the surrounding perichromosomal substances in the backscattered electron (BSE) mode at accelerating voltages of over 20kV. Findings indicated that chromatin fibers in native chromosomes have a structure similar to the hierarchic coiled model. The present study also demonstrated that not only surface structures but also subsurface structures can be studied in the BSE mode of LVSEM, when the subsurface structures have been stained with heavy metal salts such as Pt blue.

View full abstract

Nerve fibers and class II major histocompatibility complex (MHC) antigen-expressing dendritic cells have been known to gather in the dental pulp beneath carious lesions. Significant functional interactions presumably occur between the neural and immune elements. The present study analyzed the morphological relationship between class II-expressing cells and nerve fibers in fuman carious teeth, visualized by a HLA-DR monoclonal antibody and a protein gene product 9.5 (PGP 9.5) polyclonal antibody; a confocal laser scanning microscope (CLSM) and an electron microscope were used. In pulps affected by early caries, HLA-DR-positive dendritic cells aggregated mainly in the cell-free zone associated with bundles of PGP 9.5-immunoreactive nerve fibers. In pulps affected by advanced caries, the accumulated HLA-DR-positive cells and PGP 9.5-immunoreactive nerve fibers showed close association with each other especially beneath the odontoblast layer: the cells even embraced the nerve fibers. Intriguingly, class II molecules were recognized not only in dendritic cells but also in the Schwann cells of non-myelinated nerves in the pulp. Using immunoelectron microscopy, class II molecules were localized on the surface of the non-myelinating Schwann cells and also within some vesicles, whereas myelinating Schwann cells lacked this immunoreactivity. PGP 9.5-immunoreactive nerve fibers were also observed densely in the odontoblast layer, and CLSM revealed an intimate association of the nerve fibers and dendritic cells. The immunoreactivity for HLA-DR in Schwann cells depended upon the severity of the carious lesion. Class II-expressing Schwann cells are suggested to function as antigen-presenting cells in addition to dendritic cells.

View full abstract

The mature ovary contains a large number of macrophages. In the present study, the distribution of macrophages in murine ovaries at various developmental stages was immunohistochemically studied using a monoclonal antibody against F4/80, a highly specific antigen of murine macrophages. The results showed that definite F4/80-positive stains were hardly detectable in ovaries on day 0 after birth. On day 7, a few F4/80-positive cells could be identified between the developing follicles. The positive stains were irregular in shape and showed little physical contact with the primordial or primary follicles. By days 14 and 21, when the theca cell layers of growing follicles were developing, the positive cells had extended or elongated to surround the cell layer. On day 28, besides the presence of elongating positive cells surrounding the growing follicles, irregularly shaped F4/80-positive cells became apparent in the interstitium between the growing follicles and also in the capsular tissues. Thereafter, positive cells with stellate appearance were detected in the corpora lutea, which first developed around 6 weeks of age. Although the positive cells were homogenously distributed in the corpora lutea in virgin adults, only a few sporadic positive cells were found there in pregnant mice. However, the positive cells infiltrated into the corpora lutea again in the postpartum period. These results show that ovarian macrophages exhibit dramatical changes in their distribution from neonatal to postpartum periods.

View full abstract

The architecture of nitric oxide (NO)-producing nerves in the rat pylorus was studied in relation to the muscular structure. The musculature of the rat pylorus was observed to be composed of two discrete muscle loops (proximal and distal sphincters). Connective tissue septa containing neural elements divided the thick musculature of the distal sphincter into many bundles. The myenteric nerve plexus of the stomach with a subpopulation of NO-producing nerves was continuous with that of the duodenum. Nitrinergic nerve fibers which originated from the antral myenteric plexus ran through the connective tissue septa in the pyloric musculature and were densely distributed on the submucosal surface of the distal sphincter. The inner-most portion of the distal sphincter consisted of smooth muscle cells showing many cytoplasmic processes and abundant nitrinergic nerve terminals.
This particular architecture of the nitrinergic nerves in the sphincter would seem to account for the coordinate motor function of the rat pyloric sphincter.

View full abstract

To understand the developing processes of gamma-aminobutyric acid (GABA)-containing chromaffin cells and nerve fibers in the mouse adrenal gland, we examined the tissues in various postnatal stages by immunohistochemistry using a GABA antibody. From birth until postnatal week 1, GABA-immunoreactivity was seen in very few nerve fibers, and in none of the chromaffin cells. At postnatal week 2, GABA-immunoreactivity appeared weakly in clusters of chromaffin cells and strongly in relatively numerous varicose nerve fibers. The immunoreactive nerve fibers were densely distributed in the small immunonegative chromaffin cells and large ganglion cells, but only sparsely so in the weak immunoreactive chromaffin cells. At postnatal week 3, the number of the immunoreactive chromaffin cells and nerve fibers further increased compared to that at postnatal week 2. The staining pattern of GABA-immunoreactive nerve fibers in the medullas was similar to that at postnatal week 2. From postnatal week 4 until postnatal week 8, the distribution and frequency of the immunoreactive chromaffin cells and nerve fibers were also similar to those at postnatal week 3. These results suggest that the expression of GABA in the chromaffin cells and in the nerve fibers of the mouse adrenal gland may be completed by postnatal week 3.

View full abstract