Biomedical subjects
Publications and source records attributed to S Matsuo.
Vasoactive intestinal polypeptide (VIP)- and substance P (SP)-like immunoreactivities were examined in the tongue of the guinea pig by using the double immunofluorescence method. Coexistence of VIP- and SP-like immunoreactivities was suggested in many nerve fibers innervating the lingual salivary glands, as well as in a few intralingual ganglionic cells.
The incorporation of 3H-proline into the secretory ameloblasts of rat molar tooth germs cultured with or without colchicine was studied by light and electron microscope radioautography to determine the function of microtubules in the transport of precursor enamel protein from the rough-surfaced endoplasmic reticulum (rER) to the Golgi cisternae. The grain counts over the transitional vesicles, which accumulated in various cellular regions with colchicine treatment, continued to increase with chase time, unlike in controls. At 30 and 90 min chase, these counts were significantly higher than in controls. Moreover, the total grain count over the organelles (rER, pale granules, and transitional vesicles), which are positioned before the Golgi cisternae in the synthetic pathway, maintained a significantly higher level at 90 min chase in colchicine-treated tooth germs than in controls. The transport of synthesized protein to the Golgi cisternae via transitional vesicles was suppressed in colchicine-treated tooth germs. Some grains appeared with time over pale granular materials that appeared in the intercellular spaces of secretory ameloblasts with colchicine treatment. However, at each chase period, the grain count over pale granular materials was not so high as the count over the enamel in control. The present results indicate that colchicine affects the transport of newly synthesized protein from the rER to the Golgi cisterna via transitional vesicles, probably by interfering with the oriented transport related to microtubular function. It is suggested that the microtubular system may be concerned with the movement of the transitional vesicles.
The budding of small vesicles from the rough-surfaced endoplasmic reticulum (rER) was examined in the secretory ameloblast of rat molar tooth germs by ordinary fixation or prolonged osmium fixation. The budding of small vesicles from the rER was observed not only at the special region (transitional region) of the rER system, which abutted on the cis-face of the Golgi apparatus, but also at other regions of the rER in the secretory ameloblast. Small vesicles (presumed to be transitional vesicles) were adjacent to the rER, which also showed budding of vesicles. After prolonged osmium treatment, osmium deposits appeared in small vesicles, as well as in the cisternae of the cis saccule of the Golgi apparatus. Small vesicles containing osmium deposits were located at various regions of the cell, including the cis-face of the Golgi apparatus. These findings indicate that the budding of small vesicles from the rER is not restricted to the transitional region of the rER system of the secretory ameloblast, but is found at various regions of the cell. This indicates that newly synthesized proteins may be transferred from the rER cisternae to the transitional vesicles not only in the transitional region of the rER system adjacent to the Golgi apparatus, but also in other regions of the secretory ameloblast.
The distribution and origin of neurokinin A (NKA)-like immunoreactivity were investigated in feline dental pulp by an indirect immunofluorescence method. NKA-containing nerve fibres with varicosities, which entered the dental pulp via apical foramen, were distributed throughout this tissue. Many NKA-containing nerve fibres were localized around blood vessels, but some were observed apart therefrom. At the odontoblastic layer, thin NKA-containing nerve fibres were observed running straight toward the pulp-predentinal border between odontoblasts. After inferior alveolar nerve section, all NKA-containing nerve fibres disappeared in the dental pulp, while the removal of the superior cervical ganglion resulted in no change in the distribution of these fibres. The correlation of NKA-like immunoreactivity and substance P (SP)-like immunoreactivity was also investigated by double-immunofluorescence technique. The distribution of NKA-containing nerve fibres was very similar to that of SP-containing nerve fibres; it appeared that all NKA-containing nerve fibres contained SP.
Staining patterns of the surface and interior of maturing enamel of these tooth germs were examined using the glyoxal bis(2-hydroxyanil) (GBHA) solution which chelates with calcium loosely bound to hydroxyapatite to form insoluble red precipitates. An intense red, band-like pattern of GBHA staining, 1-2 mm wide, first appeared on the incisal portion of lingual enamel surface as complete loops, concentrically arranged; these gradually increased in number. Most of the later-formed bands encircled the entire periphery of the maturing enamel surface. GBHA also revealed reactive areas on the labio-lingual, cut and ground surface of maturing enamel, corresponding exactly to the surface GBHA bands. GBHA did not stain EDTA-etched surface enamel, but did reveal regular staining patterns on the ground surface, disclosed after EDTA treatment. These observations suggest an intimate correlation between the properties of interior and surface enamel, at least with regard to the state of local calcium. The maturation of bovine incisor enamel may start at the lingual aspect of the tooth crown.
The Golgi apparatus and Golgi-associated endoplasmic reticulum lysosome (GERL) were examined in the ameloblasts with a cytochemical marker, osmium impregnation, and two enzyme markers, thiamine pyrophosphatase (TPPase) and acid phosphatase (ACPase). In control cultured germs, osmium deposit appeared in one to two immature side cisternae of Golgi stacks; TPPase activity was restricted in a few mature side cisternae and condensing vacuoles. ACPase activity existed in the GERL and, sometimes, in the mature side-cisternae and condensing vacuoles. These findings show that Golgi stacks of ameloblasts consist of several distinct compartments. In colchicine-treated tooth germs, there were morphological and cytochemical changes in both Golgi stacks and GERL. The Golgi apparatus was fragmented and its stacks were scattered throughout the supranuclear region. In some stacks, the number of osmium-positive cisternae was greater than normal; in others they were absent. TPPase and ACPase activity was absent or diminished. These findings suggest the importance of microtubules in the organization of Golgi complex and GERL in the secretory ameloblast.
The glomerular distribution of type IV collagen and laminin, the major collagenous and noncollagenous components of the glomerular basement membrane, was studied by immunofluorescence microscopy in idiopathic and lupus membranous glomerulonephritis. Affinity-purified antibodies against type IV collagen reacted preferentially with the inner aspect and irregularly with the adjacent outer area of the thickened basement membrane. In contrast, laminin was detected along the inner aspect of the glomerular basement membrane, in subepithelial basement membrane protrusions ("spikes"), and in the newly formed basement membrane layer above the immune deposits. We conclude that type IV collagen and laminin do not codistribute in the newly formed matrix. This aberrant antigenic distribution may reflect a loss of coordinate biosynthesis or degradation of these matrix components by visceral epithelial cells.
Rabbits passively immunized with goat antibodies to rabbit angiotensin converting enzyme (ACE), an enzyme synthesized in the endoplasmic reticulum and mainly expressed on the apical membranes of the cells of proximal tubules, developed mild and transient immune deposits in this segment of the nephron. Granular deposits of goat IgG, rabbit ACE and C3 were found in the basolateral compartment and were maximal during the first week of immunization when the highest titers of anti-ACE antibodies were present. As the antibody titer fell to an undetectable level, the immune deposits were rapidly cleared and were virtually absent 21 days after the injections. Artificial increase of glomerular permeability allowed focal binding of ACE antibodies to the brush border of some tubules, but did not significantly alter the pattern of immune injury at the base of tubular cells. The data are consistent with the interpretation that the immune deposits result from in situ formation of immune complexes. This mechanism would involve passage of circulating antibodies across the tubular basement membrane and their combination with ACE associated with tubular cell surface membranes.
Psoriatic involved epidermis reveals variously altered receptor-adenylate cyclase responses; among them the most prominent is defective beta-adrenergic adenylate cyclase response, which is normally the major receptor-adenylate cyclase system of human epidermis. It is known that activation of hormone-stimulated adenylate cyclase, a membrane-bound enzyme complex, requires functional coupling of at least 3 distinct subunits: 1) receptor subunit (R), 2) guanine nucleotide binding protein (G), and 3) catalytic subunit (C). The precise nature of the beta-adrenergic defect in the psoriatic epidermis, however, remains to be determined, especially in terms of G and C function. Using the involved and uninvolved skin from psoriatic patients, we investigated effects of cholera toxin (which monitors G-C interaction) and forskolin (which monitors C function) on the adenylate cyclase system of epidermis, which were compared with those of normal human epidermis. Both agents increased cyclic AMP levels of involved, uninvolved, and normal human epidermis. Marked accumulations were observed in the presence of cyclic nucleotide phosphodiesterase inhibitor, isobutyl-methylxanthine (IBMX); without the phosphodiesterase inhibitor, the effect of each agent was minimal. Comparison of the effects of cholera toxin revealed that the psoriatic involved epidermis accumulates much more cyclic AMP than the uninvolved epidermis (involved: 193 +/- 65; uninvolved: 117 +/- 54 pmoles/mg protein/5 h). Similarly forskolin-induced cyclic AMP accumulations of the involved epidermis were much more than those of uninvolved epidermis (involved: 374 +/- 152; uninvolved: 101 +/- 41 pmoles/mg protein/2 h). Those of normal human epidermis were not significantly different from those of uninvolved epidermis (cholera toxin: 99 +/- 36 pmoles/mg protein/5 h; forskolin: 84 +/- 22 pmoles/mg protein/2 h). Our results indicate that G and C function and their interaction is not defective (but rather increased) in the psoriatic involved epidermis. This suggests that the defective beta-adrenergic response of psoriatic involved epidermis reflects defective R or R-G interaction of the epidermal adenylate cyclase system.
Cytodifferentiation of the chick pineal gland throughout the embryonic development was investigated with light and electron microscopy. The chick pineal anlage appears first as a small evagination in the diencephalic roof at 60 h of incubation (27-30 somites). Until day 5 of incubation, pineal anlage cells are undifferentiated and appear similar to ventricular ependymal cells. Subsequently, pinealocytes and supporting cells are first distinguishable at 7-8 days, and parafollicular cells are distinguishable at 12 days of incubation. Pigment-containing cells after 6 days and nerve cells after 17 days of incubation gradually increase, especially in the posterior wall of the pineal recess. During embryonic development, the chick pineal gland has both photosensory and secretory elements: viz. the former, mitochondria-laden apical protrusions, synaptic ribbons, lamellar whorl-like cilia of the pinealocytes, and adjacent appearance of the pigment-containing cells and the nerve cells; and the latter, dense-cored vesicles of the pinealocytes and dense bodies of the supporting cells. Moreover, nuclear invaginations having a large lipid droplet nearby and some aggregations of glycogen are found in the pinealocytes and are transitory changes in structure restricted to certain days of incubation.
Substance P-like immunoreactive (SPLI) nerve fibers were demonstrated in the Krause corpuscles of the dog's tongue using the indirect immunofluorescence method and cholinesterase histochemistry. SPLI nerve fibers were often in contact with Krause end bulbs and occasionally entered them. From this result it was suggested that substance P might be involved in sensory mechanism of the Krause apparatus.
We have identified a monoclonal antibody (ERY-1), which reacts with erythrocytes, erythroid precursor cells, and with embryonal yolk sac, and normal liver and kidney. The antibody also decorates the neoplastic cells of hepatocellular, renal, and yolk sac carcinomas. No reactivity was seen in a variety of other epithelial or mesenchymal neoplasms. It is possible that ERY-1 recognizes an erythropoiesis-associated antigen present in yolk sac, kidney, liver, and bone marrow, all of which are involved in erythropoiesis in various stages of human development. Furthermore, ERY-1 has proved to be extremely useful in the histopathologic diagnosis of hepatocellular and renal cell carcinomas.
BALB/c mice were exposed simultaneously to three nonrelated immunogens, myelin basic protein, uridyl-galactosyl transferase, and tissue obtained from a formalin-fixed, paraffin-embedded block containing a pilomatrixoma. Standard hybridoma techniques were used and antibody generation assayed using an unlabelled antibody biotin-avidin method with sections of human cerebellum, liver, and pilomatrixoma as the substrates. Using the above assay, clones were selected that secreted antibodies with selective specificities for each of the immunogens. By ELISA assay, the monoclonal antibodies reacting with cerebellum also reacted to the myelin basic protein preparation used as immunogen and the monoclonal antibody reacting to hepatocytes bound to the preparation of uridyl-galactosyl transferase used as immunogen. Our data suggests that the generation of monoclonal antibodies with a variety of diagnostic applications can be obtained from a single fusion following immunization with multiple nonrelated antigens, requiring considerably less laboratory cost and effort than would be required to obtain similar monoclonal antibodies in separate fusions.
Electrophysiologic study of a 55-year-old patient with Lown-Ganong-Levine syndrome associated with apical hypertrophic cardiomyopathy is reported. The patient had a history of recurrent attacks of tachyarrhythmia and his electrocardiogram showed a short P-R interval (0.10 sec) with narrow QRS complex and left ventricular hypertrophy with giant negative T waves. His cineangiogram showed severe apical hypertrophy. An electrophysiologic study was performed. The results of programmed atrial pacing show the existence of the dual A-V nodal pathways. The A-H interval at rapid atrial pacing increased maximally by 103 msec. Atrial stimulation could depolarize parts of the atrium without altering the supraventricular tachycardia. These findings suggested that preferential rapidly conducting A-V nodal and intranodal reentry are the responsible mechanisms in this reciprocating tachycardia. We conclude that the short P-R interval was due to intranodal reentry through the dual A-V nodal pathways. To our knowledge, a case of Lown-Ganong-Levine syndrome with apical hypertrophic cardiomyopathy has not been previously described in the literature.
The location of calcium in a rapid-frozen and freeze-substituted maturation stage enamel organ of the rat incisors was demonstrated by means of the glyoxal bis(2-hydroxyanil) (GBHA) staining method, which formed insoluble red precipitates of calcium-GBHA complex. In the ameloblast layer, highly GBHA-reactive tubulo-vesicular structures corresponding to mitochondria and some other membrane-bound structures were localized in both ruffle-ended and smooth-ended ameloblasts, although no significant GBHA reaction was localized in the nucleus, Golgi region, nor along the plasma membrane of these cells. In addition, numerous granular GBHA reactions appeared exclusively in association with the ruffled border of ruffle-ended ameloblasts. GBHA reactions were positive, but were considerably weaker in papillary cells than in the ameloblast. These observations provide a first published histochemical mapping of calcium in the maturation stage enamel organ, and suggest the active participation of mitochondria in maturation stage ameloblasts in calcium regulation.
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