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Volume 19 (1986)
- Issue 6 Pages 687-
- Issue 5 Pages 545-
- Issue 4 Pages 421-
- Issue 3 Pages 319-
- Issue 2 Pages 161-
- Issue 1 Pages 1-
Volume 19, Issue 5
Displaying 1-11 of 11 articles from this issue
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ISAO HOZUMI, TOSHIHIKO IWANAGA, TSUNEO FUJITA, TOHRU YAMAKUNI, YASUO T ...1986 Volume 19 Issue 5 Pages 545-553
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSSpot 35 protein is a cerebellar protein which is restricted, in the cerebellum, to Purkinje cells. Immunohistochemical localization of the protein shows it to occur also in peptide/amine-producing endocrine cells of guinea pigs, cats and dogs. Spot 35 protein immunoreactivity occurred in pituitary endocrine cells, carotid body chief cells, thyroid parafollicular cells, pancreatic islet cells, gut endocrine cells and adrenal chromaffin cells. In the pituitary, gut and adrenal medulla, only some endocrine cells stained positively. In the pancreas of cats and dogs, most cells of the islets showed the immunoreactivity, while in that of guinea pigs somatostatin-producing D cells were selectively immunoreactive for spot 35 protein.View full abstractDownload PDF (8868K) -
SHINICHIRO TSUYAMA, TATSUO SUGANUMA, KAORI IHIDA, FUSAYOSHI MURATA1986 Volume 19 Issue 5 Pages 555-565
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSThe mucous glycoproteins of mucous cells in the lower half of the rat proximal colon were examined ultracytochemically to study the intracellular sites of glycosylation in relation to the cell differentiation and the functional polarity of the cell organellae using a combination of a hydrophilic resin embedment and a postembedding staining technique with lectin gold com plexes.
In the immature mucous cells located near the branching point of the glandular crypt, the Golgi stacks and mucous granules were stained with soybean agglutinin (SBA), Ricinus communis agglutinin-I (RCA-I) and Helix pomatia agglutinin (HPA). In the mature mucous cells at the bottom of the crypt, the cis region of the Golgi stack was stained with SBA, a few intermediate saccules were stained with RCA-I, and both the cis and trans sides of the cisternae and mucous granules, but not the intermediate lamellae, were stained with HPA. After neuraminidase digestion of ultrathin sections, the RCA-I binding sites were greatly increased. Ulex europeus agglutinin-I (UEA-I) and Limax flavus agglutinin (LFA) positively stained the trans side lamellae and mucous granules of the mucous cell at the bottom of the glandular crypt.
The processing of the glycosylation of the mucous glycoprotein in the lower half crypt of the rat proximal colon was partly clarified electron microscopically.View full abstractDownload PDF (9984K) -
NOBORU SHINDO, MASAAKI OKADA1986 Volume 19 Issue 5 Pages 567-579
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSThis study was undertaken to reveal the correlation with immunoglobulins, complement and electron dense deposits seen by ordinary electron microscopy (EM), and to clarify whether the main site of deposition differs according to the type of immunoglobulins, using an immunoelectron microscopic (IEM) technique, with minor modification. Study cases consisted of 13 patients with various types of glomerulonephritis. Electron microscopic, and immunofluorescence (IF) examinations were performed by the standard methods. For IEM, glutaraldehyde-fixed kidney specimen was used.
In all cases, the localization of immunoglobulins and complement seen by IEM corresponded to electron dense deposits seen by ordinary EM. By IEM on various types of glomerulonephritis, the main site of deposition differed according to the type of immunoglobulins; IgG was found in all locations, while IgA and IgM were found mainly in subendothelial and mesangial areas.
In one case of membranous nephropathy and one case of IgA nephropathy, IgG was found to be outside of the glomerular basement membrane (GBM), while IgA was seen to be inside the GBM. The method described in this study can be used for examination of both ordinary EM and IEM using the same kidney tissue. This is important to make strict correlation between immunopathologic and histopathologic findings.View full abstractDownload PDF (20138K) -
SINICHIRO SUMITOMO, SHUNSUKE KUMASA, YOSHIMASA IWAI, MASAHIKO MORI1986 Volume 19 Issue 5 Pages 581-587
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSAn immunohistochemical study was undertaken to determine the localization of involucrin in human salivary gland lesions and tumors. Normal glandular tissues and those with obstructive sialadenitis were negative for involucrin staining. In pleomorphic adenomas, the luminal surface of tubular, ductal, duct-like, and cystic structures was positive for involucrin, and some cells lining the cavities of these structures were also positive. In the outer layer of the duct-like structure, spindle-like cells having long anastomosing processes were negative. Squamously metaplastic cells and cells with ongoing keratinization strongly indicated the presence of involucrin. In Warthin's tumor specimens, cells strongly positive for involucrin were found, though rarely, scattered around the eosinophilic tumor epithelium. Immunohistochemically detected involucrin may be a specific marker for detecting squamous metaplasia or keratinizing change in the epithelia of salivary gland tumors.View full abstractDownload PDF (8750K) -
I. PREPARATION AND CHARACTERISTICS OF MONOCLONAL ANTIBODIES TO γ-GLUTAMYL TRANSPEPTIDASEKENJIRO YASUDA, SHUJI YAMASHITA, SADAKAZU AISO, MASAHIDE SHIOZAWA, TOS ...1986 Volume 19 Issue 5 Pages 589-600
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSMonoclonal antibodies of the IgG1 subclass were prepared against rat kidney γ-glutamyl transpeptidase by the hybridoma technique. Eight hybrid clones produced antibodies against the enzyme. Finally two stable clones which produce specific antibodies were obtained. The antibodies specifically recognized a heavy subunit (Mr=51, 000) of the enzyme. Immunohistochemical examination revealed that the reaction was localized in the brush border as well as along the basolateral membrane of the proximal convolution of rat kidney and also in the epididymis and the pancreas of rats. Human kidney showed positive reaction, whereas no reaction was recognized in the kidneys of other species, such as mice, guinea pigs and rabbits.View full abstractDownload PDF (10705K) -
Ca++-ATPase AND Na+, K+-ATPaseKAZU SHIGE OGAWA, KAZUSHI FUJIMOTO, KAZUO OGAWA1986 Volume 19 Issue 5 Pages 601-620
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSCa++-activated adenosine triphosphatase (Ca++-ATPase) and ouabain-sensitive, K+-dependent p-nitrophenylphosphatase (K+-NPPase), a component of Na+, K+-ATPase system, were studied in rat aortic endothelium and smooth muscle cell using the recent improved cytochemical methods introduced by Ando et al. (1981) and Mayahara et al. (1980) respectively.
In the endothelium, Ca++-ATPase activity was localized on the external side of luminal, lateral and abluminal membrane, on the inside of caveolae and vesicles of both luminal and abluminal surfaces and on the matrix of mitochondria. Furthermore, Ca++-ATPase activity on only the luminal surface was affected by the concentrations of Ca++ in reaction medium. K+-NPPase activity was recognized on the cytoplasmic side of or exactly on the membrane covering abluminal caveolae, abluminal vesicles and Golgi apparatus. These findings indicate the existence of a cytochemical difference on the aortic endothelial cell-surface corresponding to the function.
On the other hand, in the smooth muscle cell, reaction products indicating Ca++-ATPase activity were localized on the plasma membrane, the caveolae, the sarcoplasmic reticulum, the Golgi apparatus and the matrix of mitochondria. K+-NPPase activity was observed on the cytoplasmic side of or just on the caveolae and the Golgi apparatus in the smooth muscle cell. These findings confirm the previously reported biochemical studies and support the mechanism of the smooth muscle relaxation in relation to the sequestration or extrusion of Ca++ and the function of adenine nucleotides and nucleosides.View full abstractDownload PDF (22268K) -
TADASHI ITOH, TOYOSHI FUJIMOTO, HIROTAMI KOIKE, TAKAO INOUE, KAZUO OGA ...1986 Volume 19 Issue 5 Pages 621-633
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSIn an attempt to detect cathodoluminescence (CL), an Akashi scanning electron microscope (SEM) ISI-DS130 was remodeled into a cathodoluminescence electron microscope (CEM or fluorescence electron microscope FEM). A newly shaped mirror, combining three half ellipsoidal mirrors, was located over the specimen holder of the top stage for color CL use. The bottom stage was modified with a half ellipsoidal mirror, photomultiplier tube (PMT) and monochrometer, so that not only CL images, but also CL spectra could be obtained. An effective specimen-preparation method, known as the simple cryo-SEM method (24), was devised for CL observation.
With these instruments and methods, color CL images and CL spectra of the biological substances were analyzed. Ito cells of vitamin A (vit A) -loaded rat liver and zona fasciculata cells of rat adrenal cortex emitted intense CL from their lipid droplets (LD). In order to identify the origin of this CL emission, analysis of the color CL images and the CL spectra were available.View full abstractDownload PDF (8732K) -
ROBERT C. ALLEN, BRUCE BUDOWLE, CALVIN A. SARAVIS, PETER M. LACK1986 Volume 19 Issue 5 Pages 637-645
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSRehydratable polyacrylamide and conventionally prepared ultrathin-layer gels (250-375μm thick) were compared. A desired ampholyte, substrate and dye, or antibody was introduced into the dried gels by rehydration. Conventional gels were used both fresh or after storage in sealed aluminized bags. The lower conductance in rehydrated gels, as compared to conventionally cast gels, allowed higher voltage gradients to be applied at controlled Joule heat loads, with increased resolution of the system. Studies with erythrocyte acid phosphate (EAP) indicated that increased activity and greater resolution were obtained in the rehydrated gels as opposed to those cast in the conventional manner. In addition, glycerol also was able to reduce conductance further, however, glycerol also reduced enzyme activity of erythrocyte acid phosphatase (EAP) C which may be caused by an isomeric conversion to EAP B in its presence. Impregnation of antibody into the gel by rehydration produced a material for immuno-sandwich overlays that allowed reaction times of as little as three minutes with focused Gc proteins to produce a specific precipitin. Unreacted antibody was washed completely from the gel in as little as 30min leaving sharply resolved precipitin lines in the gel that could be stained with both Coomassie Brilliant Blue R250 or with silver techniques. Studies of the isoelectric points by automated laser microdensitometry indicated a high degree of reproducibility between gels.View full abstractDownload PDF (5002K) -
BRUCE BUDOWLE, ANNE M. GAMBEL1986 Volume 19 Issue 5 Pages 647-654
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSA simple method was described for negative gold staining of ultrathin-layer polyacrylamide gels. The protein bands were clear (unstained) and sharp while the gel matrix was pink-purple. Negative gold staining was up to five-fold more sensitive than coomassie blue, but an order of magnitude less sensitive than silver staining for protein detection. However, there were examples where negative gold staining was able to detect particular proteins not effectively stained by coomassie blue or silver.View full abstractDownload PDF (8901K) -
CARL R. MERRIL1986 Volume 19 Issue 5 Pages 655-667
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSSilver stains permit the detection of nanogram amounts of proteins and nucleic acids in gels or membranes. These silver stains have been adapted from histological and photographic photochemical protocols. The basic mechanism of the visualization of protein and nucleic acids by silver staining involves the reduction of ionic to metallic silver. Staining properties of individual amino acids, homopolymers, and small peptides, have been used to demonstrate the importance of the basic amino acids, lysine and histidine, and the sulfur containing amino acids in the silver staining of proteins while the purines have proven to be important in the staining of nucleic acids. Many silver stains demonstrate reproducible curvilinear relationships between silver densities and protein and nucleic acid concentrations. Their sensitivity and reproducibility permits their use in quantitative analysis. By utilizing sets of operationally con stitutive proteins for the normalization of intra-gel stain intensities, quantitative comparisons of protein concentrations have been made in electrophoretograms from complex biological fluids or cellular extracts. These ultrasensitive silver stains have also permitted the discovery of disease associated spinal fluid proteins in a number of central nervous system diseases.View full abstractDownload PDF (1690K) -
TETSUJI NAGATA, TOSHIAKI NISHIGAKI, YASUNORI MOMOSE1986 Volume 19 Issue 5 Pages 669-683
Published: 1986
Released on J-STAGE: October 28, 2009
JOURNAL FREE ACCESSIn order to demonstrate the intracellular localization of anti-allergic agent in rat mast cells, 3H-labeled Tranilast, N-(3', 4'-dimethoxycinnamoyl) anthranilic acid was studied by both light and electron microscopic radio-autography.
The peritoneal mast cells of several Wistar strain rats were collected and incubated in vitro with Tranilast (100μM) after sensitization with IgE and challenging with an antigen, demonstrating the inhibition of degranulation by type I allergic reaction. Then, the peritoneal mast cells were incubated in vitro with 3H-Tranilast (100μCi/ml) for varying time intervals from 0 to 60min and the cell pellets were fixed, embedded and processed for light and electron microscopic radioautography. Light microscopic radioautograms revealed that silver grains were localized over the cytoplasm. Electron microscopic radioautograms revealed that silver grains were localized over and around the granules. The number of silver grains increased according to the prolongation of incubation time.
Several rats were administered orally with 3H-Tranilast (53μCi/gm body weight). They were sacrificed at various time intervals and the back skins were taken out, fixed, embedded and processed for light and electron microscopic radioautography. Silver grains were observed to localize over the subcutaneous tissues, especially concentrating over the mast cells.
From the results, it is concluded that 3H-Tranilast was rapidly taken into mast cells in both in vitro and in vivo and it inhibited degranulation of type I allergic reaction.View full abstractDownload PDF (11919K)
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