Cellular differentiation and histogenesis of the architecture of the temporal muscle of rats were studied by histochemical staining for SDH. From day 19 of gestation to day 16 after birth, development of two kinds of muscle fibers could be distinguished: fibers with fine formazan crystals distributed diffusely in the sarcoplasm (SS) and fibers with large formazan particles distributed diffusely in the sarcoplasm (SL). After day 16 some muscle fibers showed marked progress of cellular differentiation and became similar to adult muscle fiber types. After day 23 most muscle fibers could be classified by Stein and Padykula (55). By day 1 after birth the organization of the primordial muscle structure observed in cross section became similar to that of adult muscle and the regions with high and low percentage distribution of SL (Psl) values were distinguished. Thereafter the average Psl values of each muscle region increased, especially from day 8 to 9 in region A and from day 13 to 16 in regions C and D. On day 23 after birth the distribution patterns of percentage of Type B plus Type C (Pbc) and of Type C fibers (Pc) in cross section became similar to those in young adult muscle. Moreover, the range of the Psl, Pbc and Pc values of primary fasciculi in each region showed significant changes during postnatal development.
Lectins have been widely used as probes of the structure and organization of cell membrane saccharide-containing components. To determine the distribution of glycoconjugates in different areas of the rat kidney, seven different horseradish peroxidase-labeled lectins were used in the present study. The distribution of binding sites of several lectins in the proximal tubules was of special interest. The brush borders of the proximal tubules reacted differently to labeled lectins at each segment. This indicates that differences in carbohydrate composition of the brush border exist in the various segments. The ascending limb of Henle's loop (ALH) had strong binding for peanut agglutinin(PNA) and soybean agglutinin(SBA). Dolichos biflorus agglutinin(DBA) appears to be a good marker for distinguishing ALH from the distal convoluted tubule. DBA, SBA and PNA also serve as markers for the collecting tubules. In combination with various lectins, findings concerning heterogeneity in the distribution of the renal glycoconjugates were elucidated. In addition, the fine structural localization of glycoconjugates which react with some of the lectins was observed electron microscopically.
The alterations in noradrenaline (NA) nerve terminals in the rat vas deferens following chronic (up to 45 days) administration of reserpine (0.5mg/kg/day) and chlorpromazine (20mg/kg/day) have been investigated at the ultrastructural level. Twenty days after reserpinization small cored vesicles (SCVs) completely disappeared and following that, a synapse-like structure was noted between a NA terminal and a smooth muscle cell. Although no remarkable changes were found in SCVs even after chlorpromazine injections for 45 days, this treatment caused a similar structural alteration to that seen in the reserpinization.
The ATP-splitting enzyme activity at the neutral pH range was localized on the plasma membrane of osteoblasts as well as in lysosomes of all kinds of bone cells. The ATP-splitting enzyme activity on the plasma membrane seems to be different from the lysosomal one. The enzyme is (a) resistant to EDTA-decalcification, (b) localized on the plasma membrane of metaphyseal osteoblasts, (c) dependent on Mg++ ion, (d) active in the neutral (pH 7.0-7.4) and moderately alkaline (pH 8.5) ranges, (e) inhibited by Ca++ ion or EDTA, but (f) not inhibited by ouabain, levamisole, L-cysteine, or NaF. From these findings, it can be concluded that the enzyme activity contains a moderate amount of adenylate cyclase.
The longissimus muscle of common dolphins (Delphinus delphis) was examined by histochemical methods. The longissimus muscle had two types of myofibers: types I and II. Type I myofibers possessed strong acid-stable myosin adenosine triphosphatase (ATPase) and weak alkali-stable ATPase activity, very strong activity of NADH: Nitro BT oxidoreductase (NADHOX), and strong activity of sn-glycerol-3-phosphate: menadione oxidoreductase (GPOX). Type II myofibers possessed weak acid-stable ATPase and strong alkali-stable ATPase activity, moderately strong activity of NADHOX, and very strong activity of GPOX. The activity of 3-hydroxybutyrate: NAD+ oxidoreductase was very weak in type I and II myofibers. Type I myofibers represented about one-half the percentage of total myofibers. They contained a few fine granules of lipid pigments. The intensity of the oxidoreductase activity in the myofibers indicates that dolphin longissimus muscles possess a great capacity for oxidative and glycolytic metabolism.
Histological and histochemical alterations on gill primary lamellae of the teleost, young yellowtail (Seriola quinqueradiata), affected by the sea bloom (Chattonella antiqua) were studied and causative factors responsible for fish death were considered in the present work. A significant loss of mucous goblet cells on the afferent ridges occurred within about one hr. Among the remainder, mucous cells located on the afferent ridge of the basal part of the gill primary lamellae were markedly impaired (64% decrease in number). For TEM preparation, hyaline degenerations of the mucous cell membrane were observed. Histochemically, these mucous goblet cells contained neutral and acid glycoproteins. The cell layer on both ridges exposed to sea bloom appeared to be thinner than that of the control. The cell body of the internal multilayered mass was shrunk and intercellular spaces were markedly expanded. These edematous gill lamellae might be caused by the disappearance of the mucous coat, leading to locally impaired osmoregulations. As a result, gas exchange on the gill lamellae might be disturbed.
Monoaminergic neurons of the brain of bullfrog were studied by means of immunohistocytochemistry using antibodies to catecholamine-synthesizing enzymes and serotonin (5-HT). Monoaminergic nerve cells were similarly localized as those reported by previous histofluorescence studies, but dopaminergic neurons were for the first time identified in the pretectal area. In the preoptic recess organ, a small number of 5-HT-containing liquor contacting neurons were found to coexist with a large number of dopaminergic ones. Neurons situated in and around the nucleus tractus solitarius were partly phenylethanolamine-N-methyltransferase-positive, supporting adrenaline to be of importance for the function of the frog brain. Moreover, the liquor contacting neurons of the paraventricular organ or nucleus infundibularis dorsalis and of the floor of the fourth ventricle were recognized as serotonergic and noradrenergic in nature, respectively. The ultrastructural study has shown that tyrosine hydroxylase (TH)- and 5-HT-positive reaction products were found in the ground substance and granules. The size of TH- or 5-HT-positive granules was variable from 90-200nm. Many differences between the intrinsic neurons in the brain parenchyma and the liquor contacting neurons were found in regard to the ultrastructure such as the number of immunoreactive granules and the nucleo-cytoplasmic ratio.
Human umbilical cord blood cells were cultured in suspension for 15 days. Characteristics of the cord blood cells grown in culture were studied in relation to histochemical features with 3H-thymidine (3H-TdR) labeling in the course of the 15-day culture. The labeling index after 1 day was 2.9% of mononuclear cells, naphthol AS-D chloroacetate esterase (NACE) and peroxidase positive cells comprised about 83% of all the labeled cells. In the study with blood from 38 to 41 weeks gestation, the percentage of 3H-TdR labeled cells and histochemically stained cells were found more or less identical regardless of the gestational age. No labeled cells with nonspecific esterase activity were observed in 1 and 5-day cultures. The total cell population decreased in 5 days whereas the labeled cells increased to 19%. The labeled cells showing NACE and peroxidase activities decreased markedly after 5 days. After 10-15 days of culture, both cell number and the labeling index decreased, whereas the labeled cells with peroxidase and luxol fast blue (LFB) activity increased. At the same time, the appearance of the labeled cells having nonspecific esterase activity was observed.
Chromaffin cells in the interrenal gland of clawed toad (Xenopus laevis) were studied by a combination of fluorescence histochemistry and electron microscopy. Microspectrofluorometrical analyses indicated that there were adrenaline(A)-containing and noradrenaline(NA)-containing chromaffin cells, both of which were scattered within the interrenal tissue. In addition, it was found that there existed a third type of chromaffin cells which emitted a primary catecholamine fluorescence. This cell type could be identified electron microscopically as small granule chromaffin (SGC) cells, based on the size of secretory granules (100nm in diameter) which were considerably smaller than those that of NA (200nm) or of A (250nm) cells.
A new one-step ultracytochemical method for the detection of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase (K+-NPPase) activity of the Na+-K+-ATPase complex was developed by Mayahara et al. (7, 8). We have now applied this method to the central nervous system of the rat. K+-NPPase activity was most intense over the entire plasma membrane of the synaptic area. Reaction products showing K+-NPPase activity were also precipitated on the axolemma, dendritic plasma membrane and filamentous structures in the axon and dendrite. No significant activity was found on the glial plasma membrane.