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  • 1
    Electronic Resource
    Electronic Resource
    Translocation of pICln in a pig kidney cell line, LLC-PK1, by low osmotic stress (1999)
    Springer
    Clinical and experimental nephrology 3 (1999), S. 163-168 
    Details
    ISSN: 1437-7799
    Keywords: Key words pICln ; Chloride channel ; LLC-PK1 ; ATP ; Azide ; Dihydrocytochalasin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background. There has been no conclusive explanation regarding the function of pICln (a 26- to 27-kDa acidic protein) on an osmo-sensitive chloride channel responsible for an outwardly rectifying anion current. We observed the effects of the hypotonic treatment of LLC-PK1 cells on the intra-cellular dynamic state of pICln. Methods. LLC-PK1 cells were cultured, and pICln in cells was observed immunohistochemically. The cells were fractionated into nuclei, mitochondrial, microsomal, and soluble fractions biochemically, and pICln was detected by an immunoblotting method after sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Results. pICln in cells was observed on nuclei and their surroundings, but not on cell membranes. pICln was present in soluble and insoluble forms. The molecular masses of the oligomeric forms in the soluble fractions were different from those previously reported with Madin-Darby canine kidney (MDCK) cells, indicating the differences in the pICln-oligomer depending on cell type. On analysis with SDS-polyacrylamide gel electrophoresis, the exposure of cells to hypotonic media elevated the ratio of soluble to insoluble forms within 5 min. This result also conflicted with those previously reported with MDCK cells. This finding suggests that the function of pICln and the signaling mechanism differ depending on the cell species. Both extracellular ATP and NaN3 inhibited this elevation of the soluble/insoluble ratio, coinciding with previous reports that extracellular nucleotides and depletion of intracellular ATP inhibited the volume-sensitive chloride channel. Dihydrocytochalasin B, an F-actin-disrupting drug, inhibited the elevation of the soluble/insoluble ratio. Conclusions. The soluble form of pICln was increased within 5 min by exposure of LLC-PK1 cells to hypotonic media. This translocation was inhibited by extracellular ATP, NaN3, and dihydrocytochalasin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s101570050029
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  • 2
    Electronic Resource
    Electronic Resource
    Correlation Between the Expression of Methionine Adenosyltransferase and the Stages of Human Colorectal Carcinoma (2000)
    Springer
    Surgery today 30 (2000), S. 706-710 
    Details
    ISSN: 1436-2813
    Keywords: Key words Methionine adenosyltransferase ; Colorectal adenocarcinoma ; Colon ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Methionine adenosyltransferase (MAT) catalyzes the synthesis of S-adenosylmethionine (AdoMet) from ATP and L-methionine. AdoMet is the major methyl donor in most transmethylation reactions in vivo, and it is also the propylamino donor in the biosynthesis of polyamines. In the present study, we assessed MAT activity in human colons with colorectal carcinoma and the values were compared with those of morphologically normal adjacent mucosa. Higher levels of MAT activity were observed in the colorectal carcinoma than in the normal colon. The ratio of MAT activity in tumor tissue versus normal tissue seemed to be correlated well will the stage of the colorectal tumor. Furthermore, immunoblot analysis showed that the high levels of MAT activity observed in colorectal carcinoma were due to the increased amounts of MAT protein. Immunohistochemical analysis revealed that MAT was most abundant in goblet cells, particularly in granules in the supranuclear area of these cells. In the colorectal carcinoma tissues, MAT was strongly stained in the cancerous cells and localized in granules in the supranuclear region. The results of this preliminary study suggest that determination of the relative ratio of MAT activity in both normal and tumor regions in human colorectal carcinoma could be a clinically useful tool for determining the stage of malignancy of colorectal carcinomas.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s005950070081
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  • 3
    Electronic Resource
    Electronic Resource
    Hepatic stellate cells (vitamin A-storing cells) change their cytoskeleton structure by extracellular matrix components through a signal transduction system (1998)
    Springer
    Histochemistry and cell biology 110 (1998), S. 121-128 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  When cultured on a polystyrene surface or aminoalkylsilane-coated cover glasses, rat and human hepatic stellate cells exhibit a flattened, fibroblast-like shape with well-developed stress fibers. However, culturing the cells on type I collagen gel results in the elongation of long, multipolar cellular processes, whereas cells cultured on Matrigel maintain their round shapes. Dual fluorescence staining of microtubules and fibrillar actin indicated that the processes extend together with collagen fibers and contained microtubules as the core, whereas the periphery contained fibrillar actin. Immunofluorescence staining of vinculin showed that the focal adhesions were distributed mainly in lamellipodia when cultured on aminoalkylsilane-coated cover glasses, whereas in the cells cultured on type I collagen gel they were localized to the tips of the processes and along their bottom surface contacting collagen fibers. Wortmannin, as well as staurosporin and herbimycin A, inhibited the elongation process and induced the retraction of elongated processes. The wortmannin treatment also resulted in an alteration in focal adhesion distribution from the processes to cell bodies. These results indicate that the cell surface integrin binding to interstitial collagen fibers induces the elongation of processes through signaling events and the subsequent cytoskeleton assembly in hepatic stellate cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004180050273
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  • 4
    Electronic Resource
    Electronic Resource
    Immunocytochemical and in situ hybridization evidence for the coexistence of GABA and tyrosine hydroxylase in the rat locus ceruieus (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 234 (1992), S. 593-604 
    Details
    ISSN: 0003-276X
    Keywords: GABA ; TH ; GAD mRNA ; Cocxistence ; LC ; Rat ; Zamboni's ; ABC ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We have demonstrated the coexistence of GABA-like and tyrosine hydroxylase-like immunoreactivities (GABA-LI and TH-LI, respectively) in the same neurons of the rat locus ceruleus (LC). The profiles of these cells were labeled by alternately immunostaining adjacent sections for GABA-LI or TH-LI by the avidin-biotin-peroxidase complex method or the peroxidase-anti-peroxidase method after perfusion (either Zamboni's fixative or PPG), and observation at light and electron microscopic levels. For light microscopy, pairs of adjacent sections of more than 590 (Zamboni's) and 260 (PPG), and for electron microscopy, 40 ultrathin sections cut from adjacent semithin plastic sections (Zamboni's), were examined. GABA-LI was found in 80% (1,309/1,642 in total) of small and medium-sized neurons, uniformly scattered throughout the LC. Observations unequivocally show that the majority of GABA-ergic neurons are also noradrenergic. Several neurons are neither noradrenergic nor GABA-ergic, while other noradrenergic neurons do not show GABA-LI. It is shown that astrocytes, but not oligodendrocytes, contain GABA. In situ hybridization using a probe DNA fragment of the glutamic acid decarboxylase (GAD) cDNA, amplified by the polymerase chain reaction, detected GAD mRNA signals in many neurons throughout the LC, supporting the presence of a GAD/GABA system in the LC. Multiple “classical” transmitters, including GABA, serotonin, and noradrenaline, coexist in many LC neurons and may contribute to its widely diverging projections throughout the entire CNS.© Willey-Liss, Inc.
    Additional Material: 21 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/ar.1092340415
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