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Volume 36 (2003)
- Issue 6 Pages 495-
- Issue 5 Pages 415-
- Issue 4 Pages 249-
- Issue 3 Pages 185-
- Issue 2 Pages 99-
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Volume 36, Issue 2
Displaying 1-10 of 10 articles from this issue
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REVIEW
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Robert Yoshiyuki Osamura, Noboru Egashira, Michio Yamazaki, Syunsuke M ...Article type:
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2003 Volume 36 Issue 2 Pages 99-103
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSMany aspects of molecular mechanisms for the production of hormones have been clarified by recent, rapidly growing molecular technologies. The cloning of various transcription factors and the gene targeting technologies have contributed a great deal in studying the essential aspects of hormone production. Prohormone convertases (PCs) and the intracellular secretory mechanisms of the prohormones have been other inevitable issues in the proper function of the hormone secreting cells. In the field of pituitary research, transcription factors such as Pit-1, Ptx1, Prop-1, NeuroD1, GATA2, SF-1, and DAX-1, clarify three cell lineages in the functional development of the individual pituitary cells. In the human pituitary adenomas, occasionally their hormone production is 'aberrant', which causes overlap in the lineages. Molecular mechanisms have been proposed to be resulted from 'aberrant' combinations of the transcription factors by the gene transfection method. Histochemistry and cytochemistry are expected to play essential future roles in the research on the physiological and pathological aspects of hormone production.
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Toshiaki Nakatani, Toshikatsu Nakashima, Taizo Kita, Akihiko IshiharaArticle type:
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2003 Volume 36 Issue 2 Pages 105-114
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSMammalian skeletal muscles are comprised of heterogeneous types of fibers based on their enzyme histochemical or immunohistochemical properties. Regardless of the muscle type, muscle region, or fiber type, an inverse relationship between cross-sectional area and oxidative enzyme activity of fibers is observed in the rat hindlimb muscles including the soleus, plantaris, extensor digitorum longus, and tibialis anterior muscles. This indicates that smaller-sized fibers have higher oxidative enzyme activities than larger-sized fibers even within the same fiber type. In addition, there is a matching of cell sizes and oxidative enzyme activities of spinal motoneurons and the muscle fibers that they innervate, which is important for maintaining their functional and metabolic properties.
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TECHNICAL ADVANCEMENT
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Shingo Kamoshida, Hiroshi Matsuoka, Atsuji Matsuyama, Ryoichi Shimomur ...Article type:
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2003 Volume 36 Issue 2 Pages 115-118
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSPrevious studies have suggested that high expression of thymidylate synthase (TS) is involved in the resistance of cancer cells to 5-fluorouracil (5-FU) and its derivatives. In order to overcome the difficulty in identifying TS immunoreactivity in formalin-fixed, paraffin-embedded sections, we determined the most suitable and reproducible condition of antigen retrieval for TS localization. Antigen retrieval methods examined included 0.2% trypsin, 0.002% proteinase K, and pressure cooking in 10 mM citrate buffer, pH 6.0 and pH 7.0, and 1 mM EDTA (ethylenediaminetetraacetic acid) solution, pH 8.0. Pressure cooking with EDTA was the best choice for retrieving the antigenicity of TS. Polyclonal antibody was superior to monoclonal antibody TS106. Preabsorption test demonstrated the specificity of the immunostaining. The staining sequence using routine paraffin sections can contribute to the establishment of the prognostic indicator and optimal 5-FU dosage in a variety of 5-FU-sensitive or -insensitive malignancies.
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Yuki Tajika, Toshiyuki Matsuzaki, Takeshi Suzuki, Takeo Aoki, Haruo Ha ...Article type:
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2003 Volume 36 Issue 2 Pages 119-122
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSImmunolabeling of cultured cell lines of epithelial origin is often performed to study cell polarity. To explore the localization of cellular molecules in detail, we devised an efficient and convenient method to prepare semithin and ultrathin cryosections of cells cultured on permeable support. Multiple specimens were piled up to obtain many vertical images of cells in a single section. Freezing them in cooled n-hexane resulted in clear and homogeneous frozen specimens, from which semithin and ultrathin cryosections of epitherial cells on permeable support were easily obtained. With this method, it is possible to prepare many cryosections of good quality for histochemical examination of vertical images of cells.
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REGULAR ARTICLE
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Concetta Lipardi, Qin Wei, Bruce M. PatersonArticle type:
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2003 Volume 36 Issue 2 Pages 123-134
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSRNA silencing is a highly conserved mechanism found in both the plant and animal kingdoms that is thought to protect the genome from disruption by transposons and viral integration events. Double stranded RNAs (dsRNA) produced by transposons or replicating virus result in the production of short 21-25 nucleotide double stranded RNA molecules containing a hydroxyl group on a two base-pair 3' overhang and a 5' phosphate residue. These siRNAs are the hallmark of RNA silencing, also known as post transcriptional gene silencing (PTGS) in plants, quelling in Neurospora, and RNA interference in C. elegans, Drosophila, and Dictyostelium. Remarkably these siRNAs direct the degradation of the complementary target RNA through a complicated mechanism that is just now being understood. From our studies on RNA interference in Drosophila we propose a model in which the siRNAs interact through complementarity with the target RNA and are extended by a cellular RNA-dependent RNA polymerase (RdRP) to form a critical length of dsRNA that is subsequently degraded by RNase III-related enzymes. Here, we discuss this model and the data produced in Drosophila to support it and, in turn, this model is compared to the proposed scheme for RNA silencing in mammalian systems.
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Katsuya Hiraishi, Tatsuo Ide, Fumie Jimma, Hiroshi Ohi, Fumiko Inokuch ...Article type:
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2003 Volume 36 Issue 2 Pages 135-144
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSBasigin is a member of the immunoglobulin (Ig) superfamily which has strong homology with both Ig Vκ domain and the β-chain of the major histocompatibility complex (MHC) class II antigen. Human basigin is also known as M6 antigen, neurothelin and EMMPRIN (extracellular matrix metalloproteinase inducer). In this study, we developed a novel anti-human basigin monoclonal antibody, J-BIT-1 and used it to map out the distribution of basigin in human tissues. The specificity of J-BIT-1 was confirmed by western blot analysis and immunocytochemical techniques utilizing basigin cDNA transfected L cells. J-BIT-1 appropriated to immunohistochemical staining for formalin-fixed, paraffin-embedded tissues and methanol-fixed cells. Excellent immunostaining results were obtained with formalin-fixed tissues after the antigen retrieval procedure. Our immunohistochemical observations revealed that basigin is widely expressed in various organs, but on specific cell types, such as blood capillaries of the brain, proximal convoluted tubules of the kidney, cardiac muscle of the heart, trophoblasts of the placenta and basal cells of the stratified squamous epithelium. The location of basigin is the cell surface, especially the site of cell adhesion. These observations suggest that basigin might be involved in basic cell functions which are promoted with cell-cell and cell-extracellular matrix interaction.
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Chikako Kinoshita, Takeshi Yaoi, Kumie Nojima, Shinji FushikiArticle type:
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2003 Volume 36 Issue 2 Pages 145-151
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSWe report here the effects of heavy ion beams on postnatal mouse cerebellar development, with reference to cell death. Eight-day-old B6C3F1 mice were irradiated with single doses of 0.1, 0.25, 0.5, 1.0, and 2.0 Gy, using a carbon beam of 290 MeV delivered from a heavy ion medical accelerator in Chiba (HIMAC). To compare the effects of X-rays with those of accelerated carbon ions, 8-day-old mice were exposed to X-rays single doses of 0.1, 0.25, 0.5, 1.0, and 2.0 Gy, respectively. Pups were fixed at 1, 6, 12 and 24 hr after exposure to HIMAC beams or X-rays. Four-μm-thick parasagittal sections of the cerebella were processed for hematoxylin-eosin staining as well as for staining with the TUNEL (terminal dUTP nick-end labeling) technique. The density of fragmented nuclei in the external granular layer increased with time, peaking at 6 hr after exposure, in both the HIMAC and X-irradiated groups. In the HIMAC groups, the density was significantly higher in those animals exposed to 0.25 Gy or more compared to 0 Gy, whereas in the X-irradiated groups it was significantly higher in those mice exposed to 0.5 Gy or more. Electron microscopic examinations revealed chromatin condensation in the cell nuclei in the HIMAC groups. This is the first in vivo evidence that apoptotic cell death is induced in developing mouse cerebellum after exposure to heavy ion particles. The difference in the frequency of dying cells between exposure to heavy ion particles and to X-rays may reflect the high linear energy transfer (LET) associated with a heavy ion beam.
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Kazuhisa Hasui, Fang Li, Xin-Shan Jia, Masanori Nakagawa, Takao Nakamu ...Article type:
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2003 Volume 36 Issue 2 Pages 153-164
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSAnti-Helicobacter pylori (HP) therapy succeeded in regressing gastric B-cell lymphomas (gBL) with a close relation to infestation of HP, although the exact mechanism of anti-HP therapy in gBL has not yet been clarified. In order to see a part of the mechanism, this study analyzed stromal cells in 34 gBL cases comprised of 11 cases of mucosa-associated lymphoid tissue (MALT) type and 24 cases of diffuse large B-cell lymphoma (DLBL) by means of immunohistochemistry of CD3, CD5, CD79a, CD68, CD21, S100 protein, thymidine phosphorylase (TP) and inducible nitric oxide synthase (iNOS). Numerous CD68-positive stromal cells were seen in 9 cases of MALT type and 22 cases of DLBL. Many dendritic cells (DC) expressed TP and formed an ill-defined meshwork in the background in 8 cases of MALT type and 17 cases of DLBL. In most cases, transition of CD68-positive stromal cells to DC expressing TP was recognized. There was more or less intermingling of CD3-positive T-cells in all cases. Expression of iNOS was seen in some stromal cells and glandular epithelial cells, but only in 3 cases of MALT type. In the germinal center (GC) colonization of MALT type there were CD21-positive follicular DC, CD68-positive stromal cells, TP-expressing DC and iNOS-expressing cells; whereas in that of DLBL the meshwork of CD21-positive follicular DC was destroyed and the stromal cells did not express TP or iNOS. Then, a peculiar co-existence of intermingling T-cells, CD68-positive stromal cells, DC expressing TP forming an ill-defined meshwork, and lymphoma cells were recognized in gBL. A small number of stromal cells and glandular epithelial cells expressed iNOS and prepared a nitric oxide-rich microenvironment for growth and transformation of MALT type lymphoma cells. This peculiar histogenesis of gBL was thought to explain a part of the mechanism of the anti-HP therapy against gBL.
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Takeo Aoki, Shin-ya Kogure, Hiroshi Kogo, Masami Hayashi, Yoshiko Ohno ...Article type:
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2003 Volume 36 Issue 2 Pages 165-171
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSVarious membrane molecules are sequestered to caveolae when cross-linked by antibodies or other multivalent ligands. One of the molecules, β2-microglobulin (β2m) in human fibroblasts, is known to line-up along actin filaments when bound with antibodies. In the present study we questioned whether other molecules show a similar distributional change before sequestration to caveolae. To compare with the anti-β2m antibody, two probes were used: cholera toxin B subunit (CTX-B), which binds to gangliosideGM1 , and biotinylated and nicked θ-toxin (BC θ), which recognizes raft cholesterol. BCθ showed a tendency to line-up longitudinally, which was similar to, but less obvious than the anti-β2m antibody. In contrast, CTX-B did not show any sign of linear arrangement. Despite the difference, the anti-β2m antibody and CTX-B eventually concentrated to caveolae and were incorporated into the same endo-lysosomal compartment. The result suggests that cross-linked plasma membrane molecules take two different routes to caveolae, one along actin filaments and the other independent of them.
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Tomoko Nakazawa, Shunichi Morikawa, Megumi Nishikawa, Taichi Ezaki, Ei ...Article type:
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2003 Volume 36 Issue 2 Pages 173-178
Published: 2003
Released on J-STAGE: June 05, 2003
JOURNAL FREE ACCESSPerivascular macrophages (Mato's perivascular cells) are located along microvessels in the Virchow-Robin space in mammals. We have isolated and cultured perivascular macrophages from microvessels in the cerebrum of Wistar strain male rats. The cultured cells were identified as perivascular macrophages in vivo. Polyclonal antibodies were produced by immunizing rabbits with cultured perivascular rat macrophages as antigens. The antibodies were evaluated by the immuno-fluorescence technique. These polyclonal antibodies reacted with perivascular macrophages, but not with perivascular smooth muscle cells or pericytes. The antibodies are, therefore, expected to be of use in studies on the immunological significance of perivascular macrophages, both in vivo and in vitro.
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