Torao Yamamoto

Monoamine-containing granulated cells in the frog lung

SummaryThe epithelium of the primary bronchus of the frog lung has been studied by fluorescence and electron microscopy. Clusters of five to ten, ovoid, brilliantly yellow fluorescent cells were observed in the basal portion of the epithelium. These cells contained numerous electron-dense granules of variable shape and size. The granules gave a positive argentaffin reaction at the ultrastructural level, suggesting a possible existence of monoamines in the granules. In addition, synaptic contact between the intraepithelial nerves and the cells, which was characterized by the aggregation of the granules toward the presynaptic membrane thickening of the cell, was also noted. These data are discussed in relation to similar studies in birds and mammals, and a possible function of these cells suggested.

Intracellular transport of horseradish peroxidase in the absorptive cells of goldfish hindgut in vitro, with special reference to the cytoplasmic tubules

SummaryThis study was undertaken to determine whether the numerous cytoplasmic tubules (CT) in the apical cytoplasm of goldfish hindgut absorptive cells are directly involved in the endocytotic transport of macromolecules into the cells, or whether they are derived from the intracellular membrane components. The absorptive cells were exposed to horseradish peroxidase (HRP)-containing medium in organ culture and subsequently fixed and prepared for electron microscopy. Analysis revealed that 5 sec after exposure, many vesicular structures, including coated vesicles, were labelled with reaction product whereas almost all CT were negative. After a 1-min exposure, reaction product was detected in about 11 % of the CT, and thereafter, the percentage increased to about 95% after 15 min exposure. As labelled CT increased in number, the number of densely labelled vacuoles with attached CT also increased. CT connected to vacuoles with a peripheral margin of dense reaction product were always HRP-positive, whereas those connected to vacuoles which were not distinctly labelled were themselves also devoid of HRP reaction product. This indicated that the labelling of CT was closely associated with the labelling of the inner surface of the vacuolar membrane. These results indicate that CT are probably formed by a budding off from these vacuoles, rather than being directly involved in endocytosis.

Immunohistochemical localization of 14 kDa β-galactoside-binding lectin in various organs of rat

SummaryImmunohistochemical localization of 14 kDa β-galactoside-binding lectin in various organs of adult rat was achieved using a monospecific antibody raised against lectin purified from rat lung. The antibody-stained cells were formed into small aggregates, thin fascicles, or thick bundles in the walls of blood vessels, gastrointestinal tracts and urogenital organs. From the patterns of distribution, as well as their organization, these immunoreactive cells were regarded as smooth muscle cells. This was confirmed by a double immunofluorescence study using a mixture of anti 14 kDa lectin and anti α-smooth muscle-specific actin antibodies. Strong 14 kDa lectin immunoreactivity was seen in the pericellular matrix of smooth muscle cells in intact organs as well as in detergent-treated organs from which all cellular components were extracted. From these findings, it is suggested that the 14 kDa lectin may be externalized by smooth muscle cells into their pericellular matrix and participate in the crosslinking of the complementary glycoconjugate(s) localized at that site. The macromolecular complex of glycoconjugates thus formed around smooth muscle cells may play a role in anchoring smooth muscle cells to the pericellular connective tissue thereby permitting the force of muscle contraction to be efficiently transmitted to the surrounding connective tissue proper.

APUD-type recepto-secretory cells in the chicken lung

SummaryThe epithelium of the intrapulmonary airways of the chicken lung has been studied by fluorescence and electron microscopy. Numerous intensely yellow-fluorescent cells occur in the epithelium of the primary and secondary bronchi. The cell cytoplasm contains characteristic granular vesicles with an electron-dense central core. The vesicles react positively to chromaffm and argentaffin treatment, indicating that they are possible storage sites for amines. Synapse-like junctions occur between the granular cells and the intraepithelial nerve endings, filled with numerous mitochondria, suggesting that these granular cells may have a dual function as both receptor and endocrine cell.

Freeze-fracture studies of gap junctions in vertebrate cardiac muscle cells

The fine structure of gap junctions in the cardiac muscle cells of the adult rat, sparrow, snake, and lizard was examined by using freeze-fracture technique. Quantitative measurements revealed that the average areas of gap junction maculae were as small as 0.015 μm2 in the sparrow and 0.023 μm2 in the lizard, whereas a wide range of size variations from 0.0014 to 7.7 μm2 was observed in the rat. Atypical gap junctions which were composed of membrane particles arranged in a line or circle were frequently encountered in the snake, and occasionally in the lizard and sparrow. Furthermore, solitary particles showing a central dot similar to the gap junctional particles could be observed in the lower vertebrate hearts. It could be speculated that these various configurations of gap junction might represent different stages of development in the vertebrate myocardium. The functional significance of such gap junction organizations was also discussed.

Smooth muscle cell proliferation in the rat coronary artery induced by vitamin D

Ultrastructural changes induced in rat coronary arteries by oral administration of a large dose of vitamin D2 were examined by transmission and scanning electron microscopy. Our observations were that smooth muscle cell proliferation occurred without endothelial denudation and without exposure of the subendothelium. While the endothelial cells did show evidence of degeneration, the adjoining endothelial cells migrated beneath so that these degenerated cells could become detached without exposing the subendothelium. Therefore, factors other than platelets seem to participate in the phenotypic modulation and proliferation of the smooth muscle cell. Medial degeneration may be a prerequisite for formation of the lesion.

Fine structure and cytochemistry of specific granules in the lamprey atrium

SummaryFine structural and cytochemical studies were performed to examine the nature of three types of specific granules found in the atrium of lamprey; specific granules of the atrial muscle cell (ASG), interstitial cell granules (ICG) and endocardial endothelial granules (ESG).Ultrastructurally, ASG and ICG appeared quite similar in size, shape and electron opacity, while ESG were much larger and less dense in opacity than the other two.None of the granules showed positive DAB reaction or acid phosphatase reaction. Only ICG revealed positive chromaffin reaction, which agreed with formaldehyde induced green fluorescence along the atrial lumen. Phosphotungstic acid at low pH stained ICG and ASG strongly positive, and ESG weakly positive. Pronase treatment in Epon sections for 24h digested ASG alone, whereas in glycol-methacrylate embedded sections, ESG were digested first, ASG were digested thoroughly after 30 min, but ICG were not digested completely after 90 min.From these results it can be concluded that the three types of specific granules have different constituents. ESG consist of protein with some polysaccharides; ASG are composed of protein carbohydrate complexes and lack catecholamines; ICG contain catecholamine as well as protein carbohydrate complexes.

Three-dimensional architecture of elastic tissue in athero-arteriosclerotic lesions of the rat aorta☆

The overall three-dimensional architecture of elastic tissue in athero-arteriosclerotic lesions of the ascending aorta of the rat was studied, using scanning electron microscopy (SEM), after hot-formic acid extraction followed by a freeze-drying method. The lesions were induced by feeding the rats a diet containing 2% cholesterol for 6 weeks and administering massive doses of vitamin D2 for the first 4 days. SEM revealed the synthesis of elastins mainly in the intima and the degradation in the internal elastic lamina (IEL) and medial concentric elastic lamellae. Intimal elastic elements involved in atherosclerotic lesions were composed of an intricately tangled network of fibrous elastins or a row of elastic laminae that surrounded many rectangular compartments, which were separated by fenestrated septum-like elastins and were assumed to be the sites of smooth-muscle-cell proliferation. These observations indicated remodelling of the elastic laminae in the intima. Degradation in the IEL and media was attributed to either vitamin D2 intoxication, which was characterized by the formation of a large degraded portion, or an enhanced elastolytic process. These alterations of elastic elements probably reflect the exact frame of the modes of migration of aortic smooth muscle cells in the development of athero-arteriosclerotic lesions, thereby indicating the role of smooth muscle cells in the progress of atherosclerosis.

Double‐axis rotary replication for deep‐etching

We have developed a simple new method of rotary shadowing, double‐axis rotary replication, for observing three‐dimensional structures in deep‐etched, rapid‐frozen tissues. The technical details of this method are described and compared with the conventional fixed‐angle rotary shadowing procedure.

Cytoskeletal organization in the supporting cell of the guinea pig organ of Corti

The rapid-freeze, deep-etch method was used to visualize the three-dimensional organization of cytoskeletons in the supporting cells of the guinea pig organ of Corti. Deep-etched replicas showed that both the head and basal portions of the pillar cell were composed of a filamentous network consisting of several kinds of fibrous elements, into which numerous microtubules and actin filaments were tightly inserted. Myosin S1-decoration showed that the main constituent element in such filamentous networks in the basal portion of the pillar cell was the actin and tiny cross-bridges interconnected the randomly oriented adjacent actin filaments.