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  • 1
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Ubiquitin (Ub) ligation is implicated in active protein metabolism and subcellular trafficking and its impairment is involved in various neurologic diseases. In rat brain, we identified two novel Ub ligases, Momo and Sakura, carrying double zinc finger motif and RING finger domain. Momo expression is enriched in the brain gray matter and testis, and Sakura expression is more widely detected in the brain white matter as well as in many peripheral organs. Both proteins associate with the cell membranes of neuronal and/or glial cells. We examined their Ub ligase activity in vivo and in vitro using viral expression vectors carrying myc-tagged Momo and Sakura. Overexpression of either Momo or Sakura in mixed cortical cultures increased total polyubiquitination levels. In vitro ubiquitination assay revealed that the combination of Momo and UbcH4 and H5c, or of Sakura and UbcH4, H5c and H6 is required for the reaction. Deletion mutagenesis suggested that the E3 Ub ligase activity of Momo and Sakura depended on their C-terminal domains containing RING finger structure, while their N-terminal domains influenced their membrane association. In agreement, Sakura associating with the membrane was specifically palmitoylated. Although the molecular targets of their Ub ligation remain to be identified, these findings imply a novel function of the palmitoylated E3 Ub ligase(s).
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1460-9568
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: In weaver mutant mice, substitution of an amino acid residue in the pore region of GIRK2, a subtype of the G-protein-coupled inwardly rectifying K+ channel, changes the properties of the homomeric channel to produce a lethal depolarized state in cerebellar granule cells and dopaminergic neurons in substantia nigra. Degeneration of these types of neurons causes strong ataxia and Parkinsonian phenomena in the mutant mice, respectively. On the other hand, the mutant gene is also expressed in various other brain regions, in which the mutant may have effects on neuronal survival. Among these regions, we focused on the pontine nuclei, the origin of the pontocerebellar mossy fibres, projecting mainly into the central region of the cerebellar cortex. The results of histological analysis showed that by P9 the number of neurons in the nuclei was reduced in the mutant to about one half and by P18 to one third of those in the wild type, whereas until P7 the number were about the same in wild-type and weaver mutant mice. Three-dimensional reconstruction of the nuclei showed a marked reduction in volume and shape of the mutant nuclei, correlating well with the decrease in neuronal number. In addition, DiI (a lipophilic tracer dye) tracing experiments revealed retraction of pontocerebellar mossy fibres from the cerebellar cortex after P5. From these results, we conclude that projecting neurons in the pontine nuclei, as well as cerebellar granule cells and dopaminergic neurons in substantia nigra, strongly degenerate in weaver mutant mice, resulting in elimination of pontocerebellar mossy fibres during cerebellar development.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1600-079X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The expression of hydroxyindole-O-methyltransferase (HIOMT), an enzyme catalyzing the final step of melatonin biosynthesis, was examined in three pineoblastomas and five pineocytomas by in situ hybridization analysis. Distinct hybridization signals for HIOMT mRNA, though weaker than in normal pineal gland pinealocytes, were detected in two of the three pineoblastoma and three of the five pineocytoma cases. Of the pineoblastomas, hybridization signals were observed in most tumor cells of one case, while in another, signals were detected in occasional cells clustered or scattered throughout the neoplastic field. Of the pineocytomas, signals were detected in most tumor cells of two cases, while in one case, signals were detected only in occasional cells. Among these specimens, one pineoblastoma and one pineocytoma were also analyzed using northern blot and reverse transcription polymerase chain reaction (RT-PCR) analyses. In the northern blot analysis, an apparently single band corresponding to the size of HIOMT mRNA was detected in both pineoblastoma and pineocytoma RNA blots. In the RT-PCR analysis, three species of HIOMT mRNA generated via alternative splicing were detected in both tumors. These results suggest that the neoplastic cells of pineoblastomas and pineocytomas often retain the ability to express HIOMT mRNA, as in normal pinealocytes, and that HIOMT is a useful tumor marker for the diagnosis of pineal parenchymal tumors.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1440-1797
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Summary: We determined the distribution of Spot 35-calbindin-D28K, a vitamin-D dependent calciumbinding protein, in rat kidney using histochemical methods and compared it with the distribution of Ca2+-ATPase activity. Spot 35-calbindin-D28K immunoreactivity was localized in the cytosol of urinary epithelial cells in distal convoluted tubules (DCT), connecting tubules (CNT) and cortical collecting ducts (CCD), identifying the physiologically confirmed site of active transcellular calcium transport. In the cytosol, the immunoreactivity was clustered near the luminal plasma membrane and around the mitochondria. These findings indicated that Spot 35-calbindin-D28K seemed to have a cytosolic calcium buffering effect in the urinary tubular epithelial cells. Enzyme histochemical analysis showed that Ca2+-ATPase activity was localized at the basolateral plasma membrane of distal nephron segments and was strongest at the cortical thick ascending limb of Henle (CTAL), including the macula densa portion. Ca2+-ATPase activity was not evident in DCT, CNT or CCD. Strong Ca2+-ATPase activity and Spot 35-calbindin-D28K immunoreactivity did not coexist in a urinary tubular cell.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1600-079X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Feng X-L, Usui H, Fujita T, Ichikawa T, Katagiri T, Washiyama K, Kumanishi T. Postnatal developmental changes in NSE and NNE mRNA expression in the rat pineal gland: In situ hybridization histochemistry. J. Pineal Res. 1998; 24:108–116. © Munksgaard, Copenhagen〈section xml:id="abs1-1"〉〈title type="main"〉AbstractBy in situ hybridization, neuron-specific enolase (NSE) and non-neuronal enolase (NNE) mRNAs were examined in the rat pineal gland at the postnatal developmental and adult stages. The distributions of hybridized signals were analyzed in comparison with immunohistochemical staining of synaptophysin (SYN), which is a marker for pinealocytes. In SYN-positive areas that were observed throughout postnatal developmental and adult stages, we detected both NSE and NNE signals, which increased simultaneously during early postnatal development and thereafter became stationary. Quantitative analysis revealed that NNE signals were 2- to 3-fold greater in number than NSE signals at any given stage. This predominant expression pattern of NNE differed from that in neurons, which usually showed both signals at similar levels and seemed to reflect the difference in physiological function from neurons. During the early postnatal stages, a cord-like arrangement of cells without distinct SYN staining was observed. This arrangement was the thickest at postnatal day 0 and became dispersed and thinner with development, showing a relationship with formation of vascularized connective tissue stroma. By in situ hybridization, many of the constituent cells showed weak NNE signals but no distinct NSE signals. However, some cells also showed weak NSE signals, suggesting heterogeneity of these cells. The characteristic NSE and NNE expression patterns in the pineal gland cells clarified in this study might provide a basis for further studies of the differentiation and function of the pineal gland.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2307
    Keywords: Basement membrane ; Capillary growth ; Seamless endothelial cell ; Regenerating capillary ; Experimental cerebral infarction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Ultrastructural analysis of capillary changes during the repair process of experimental cerebral infarction induced in rats was carried out with special reference to the endothelial basement membrane (BM) and seamless-type endothelial cells. Following degeneration of endothelial cells and pericytes, their BMs, without any interruption or fragmentation, were left in the lesion. Newly formed capillaries grew from vessels in the surrounding brain tissues into the reactive zone of infarcts. While the capillaries in cross-section possessed multilayered BMs, these membranes in tangential section comprised an outer BM with extremely wavy profile and an inner one showing a normal trilayered structure, uniformly enveloping the endothelial surface. It is therefore suggested that the sprouting of regenerating capillaries might invade the remaining cavities of BM, resulting from endothelial degeneration. In these new vessels, seamless-type endothelial cells lacking interendothelial contacts were observed frequently. These two different and previously unobserved findings appear to be at the heart of the regeneration mechanism of reactive capillary proliferation.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 48 (1979), S. 161-163 
    ISSN: 1432-0533
    Keywords: SSPE ; Measles virus ; Immunoperoxidase method ; Encephalitis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We used the indirect immunoperoxidase method to study formalin-fixed, paraffin-embedded brain sections of four autopsied subacute sclerosing panencephalitis (SSPE) patients. Specific reaction with measles virus antigens was noted in all cases. Positive staining was observed in neurons and glia cells and both nucleus and cytoplasm were usually stained in these positive cells. Electron microscopy revealed intranuclear and intracytoplasmic nucleocapsids stained with electron-dense reaction products. Our findings suggest that for immunoperoxidase studies of SSPE formalin-fixed, paraffin-embedded materials can be used.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-0533
    Keywords: Key words: Glioma – Brain tumor – p53 gene mutations – Tumor suppressor gene – Polymerase chain reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract. Using polymerase chain reaction-assisted single-strand conformation polymorphism (PCR-SSCP) and immunohistochemical analyses, mutations in the p53 tumor suppressor gene were examined in 19 low- and high-grade gliomas. By PCR-SSCP and nucleotide analyses, p53 gene mutation was seen in 7 gliomas. Out of the 7 mutations, 3 were located at the CpG site of the previously proposed hot-spot codons 248 and 273, 2 were at codons 171 and 214 and the other 2 were in intron 5, 1 at the splice acceptor site and the other in the vicinity of the splice donor site. The latter 4 mutations have not, or only rarely, been observed in gliomas or in other tumors. However, their effect on the structural and functional alteration of the p53 protein was suggested by positive intranuclear p53 immunostaining in neoplastic cells in 3 mutations including the 1 at the splice acceptor site. In connection with glioma grading, the p53 gene mutation was shown to have occurred in both low- and high-grade gliomas, often in most of the neoplastic cells, as suggested by lack of distinct normal bands and ladders in SSCP and direct sequencing, respectively. The absence of recurrence and malignant transformation over a considerably long postoperative time in our low-grade glioma cases suggested that the p53 gene mutation might not be sufficient for the progression from low- to high-grade gliomas. The frequency of detection of mutation was 7/19 (37%) by PCR-SSCP, 8/19 (42%) by immunohistochemistry and 10/19 (53%) by both methods. The results of PCR-SSCP and immunohistochemistry were consistent in 14 cases (73.7%), but not in 5 cases (26.3%). Thus, the use of both methods was recommended to survey the occurrence of p53 gene mutation more accurately.
    Type of Medium: Electronic Resource
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