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  • Articles: DFG German National Licenses  (2)
  • Cocxistence  (1)
  • LLC-PK1  (1)
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  • Articles: DFG German National Licenses  (2)
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Years
  • 1
    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
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  • 2
    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
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