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  • 1
    Electronic Resource
    Electronic Resource
    Solubilization and Characterization of Calcitonin Gene-Related Peptide Binding Site from Porcine Spinal Cord (1988)
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 50 (1988), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The binding site for calcitonin gene-related peptide (CGRP) was solubilized with 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate (CHAPS) in an active form from porcine spinal cord. l25I-labeled human α-CGRP (125I-CGRP) binding to the solubilized protein was determined by filration using a GF/B glass filter. The maximal binding activity (˜60% of the crude membrane fraction) was obtained with 5 mM CHAPS. 125I-CGRP binding to the solubilized protein was of high affinity, saturability, and high specificity, having KD and Bmax values of 3.69 pM and 338 fmol/mg of protein, respectively. The binding activity was eluted in a single peak with a molecular mass of 400,000 daltons by gel filtration on TSK gel G4000SW. These results suggest that the solubilized protein may be responsible for the specific binding site.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1471-4159.1988.tb02936.x
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  • 2
    Electronic Resource
    Electronic Resource
    Proteolytic degradation of glutamate decarboxylase mediates disinhibition of hippocampal CA3 pyramidal cells in cathepsin D-deficient mice (2005)
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 94 (2005), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Although of clinical importance, little is known about the mechanism of seizure in neuronal ceroid lipofuscinosis (NCL). In the present study, we have attempted to elucidate the mechanism underlying the seizure of cathepsin D-deficient (CD–/–) mice that show a novel type of lysosomal storage disease with a phenotype resembling late infantile NCL. In hippocampal slices prepared from CD–/– mice at post-natal day (P)24, spontaneous burst discharges were recorded from CA3 pyramidal cells. At P24, the mean amplitude of IPSPs after stimulation of the mossy fibres was significantly smaller than that of wild-type mice, which was substantiated by the decreased level of γ-aminobutyric acid (GABA) contents in the hippocampus measured by high-performance liquid chromatography (HPLC). At this stage, activated microglia were found to accumulate in the pyramidal cell layer of the hippocampal CA3 subfield of CD–/– mice. However, there was no significant change in the numerical density of GABAergic interneurons in the CA3 subfield of CD–/– mice at P24, estimated by counting the number of glutamate decarboxylase (GAD) 67-immunoreactive somata. In the hippocampus and the cortex of CD–/– mice at P24, some GABAergic interneurons displayed extremely high somatic granular immunoreactivites for GAD67, suggesting the lysosomal accumulation of GAD67. GAD67 levels in axon terminals abutting on to perisomatic regions of hippocampal CA3 pyramidal cells was not significantly changed in CD–/– mice even at P24, whereas the total protein levels of GAD67 in both the hippocampus and the cortex of CD–/– mice after P24 were significantly decreased as a result of degradation. Furthermore, the recombinant human GAD65/67 was rapidly digested by the lysosomal fraction prepared from the whole brain of wild-type and CD–/– mice. These observations strongly suggest that the reduction of GABA contents, presumably because of lysosomal degradation of GAD67 and lysosomal accumulation of its degraded forms, are responsible for the dysfunction of GABAergic interneurons in the hippocampal CA3 subfield of CD–/– mice.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1471-4159.2005.03250.x
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  • 3
    Electronic Resource
    Electronic Resource
    Characterization of Cln3p, the gene product responsible for juvenile neuronal ceroid lipofuscinosis, as a lysosomal integral membrane glycoprotein (2003)
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 87 (2003), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Juvenile neuronal ceroid lipofuscinosis (JNCL) is an autosomal recessively inherited lysosomal storage disease involving a mutation in the CLN3 gene. The sequence of CLN3 was determined in 1995; however, the localization of the CLN3 gene product (Cln3p) was not confirmed. In this study, we investigated endogenous Cln3p using two peptide antibodies raised against two distinct epitopes of murine Cln3p. Identification of the liver 60 kDa protein as Cln3p was ascertained by amino acid sequence analysis using tandem mass spectrometry. Liver Cln3p was predominantly localized in the lysosomal membranes, not in endoplasmic reticulum (ER) or Golgi apparatus. As the tissue concentration of brain Cln3p was much lower than that of liver Cln3p, it could be detected only after purification from brain extract using anti-Cln3p IgG Sepharose. The apparent molecular masses of liver Cln3p and brain Cln3p were determined to be about 60 kDa and 55 kDa, respectively. Both brain and liver Cln3p were deglycosylated by PNGase F treatment to form polypeptides with almost the same molecular mass (45 kDa). However, they were not affected by Endo h treatment. In addition, it was also elucidated that the amino terminal region of Cln3p faces the cytosol.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1471-4159.2003.02132.x
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  • 4
    Electronic Resource
    Electronic Resource
    Identification of Cellular Compartments Involved in Processing of Cathepsin E in Primary Cultures of Rat Microglia (1998)
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 70 (1998), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Cathepsin E is a major nonlysosomal, intracellular aspartic proteinase that localizes in various cellular compartments such as the plasma membrane, endosome-like organelles, and the endoplasmic reticulum (ER). To learn the segregation mechanisms of cathepsin E into its appropriate cellular destinations, the present studies were initiated to define the biosynthesis, processing, and intracellular localization as well as the site of proteolytic maturation of the enzyme in primary cultures of rat brain microglia. Immunohistochemical and immunoblot analyses revealed that cathepsin E was the most abundant in microglia among various brain cell types, where the enzyme existed predominantly as the mature enzyme. Immunoelectron microscopy studies showed the presence of the enzyme predominantly in the endosome-like vacuoles and partly in the vesicles located in the trans-Golgi area and the lumen of ER. In the primary cultured microglial cells labeled with [35S]methionine, 〉95% of labeled cathepsin E were represented by a 46-kDa polypeptide (reduced form) after a 30-min pulse. Most of it was proteolytically processed via a 44-kDa intermediate to a 42-kDa mature form within 4 h of chase. This processing was completely inhibited by bafilomycin A1, a specific inhibitor of vacuolar-type H+-ATPase. Brefeldin A, a blocker for the traffic of secretory proteins from the ER to the Golgi complex, also inhibited the processing of procathepsin E and enhanced its degradation. Procathepsin E, after pulse-labeling, showed complete susceptibility to endoglycosidase H, whereas the mature enzyme almost acquired resistance to endoglycosidases H as well as F. The present studies provide the first evidence that cathepsin E in microglia is first synthesized as the inactive precursor bearing high-mannose oligosaccharides and processed to the active mature enzyme with complex-type oligosaccharides via the intermediate form and that the final proteolytic maturation step occurs in endosome-like acidic compartments.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1471-4159.1998.70052045.x
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  • 5
    Electronic Resource
    Electronic Resource
    Parkin is associated with cellular vesicles (2001)
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 78 (2001), S. 0 
    Details
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: We recently identified a novel gene, parkin, as a pathogenic gene for autosomal recessive juvenile parkinsonism. Parkin encodes a 52-kDa protein with a ubiquitin-like domain and two RING-finger motifs. To provide a insight into the function of parkin, we have examined its intracellular distribution in cultured cells. We found that parkin was localized in the trans-Golgi network and the secretory vesicles in U-373MG or SH-SY5Y cells by immunocytochemical analyses. In the subsequent subcellular fractionation studies of rat brain, we showed that parkin was copurified with the synaptic vesicles (SVs) when we used low ionic conditions throughout the procedure. An immunoelectromicroscopic analysis indicated that parkin was present on the SV membrane. Parkin was readily released from SVs into the soluble phase by increasing ionic strength at neutral pH, but not by a non-ionic detergent. To elucidate its responsible region for membrane association, we transfected with green fluorescent protein-tagged deletion mutants of parkin into COS-1 cells followed by subcellular fractionation. We demonstrated the ability of parkin to bind to the membranes through a broad region except for the ubiquitin-like domain. The significance of SV localization of parkin is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1471-4159.2001.00364.x
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  • 6
    Electronic Resource
    Electronic Resource
    Abnormal expression of neurofilament proteins in dysmyelinating axons located in the central nervous system of jimpy mutant mice (1999)
    Oxford, UK : Blackwell Science Ltd
    European journal of neuroscience 11 (1999), S. 0 
    Details
    ISSN: 1460-9568
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Myelination in the peripheral nervous system is considered to increase the phosphorylation level of neurofilament proteins in the axon, resulting in an increase in axonal calibre. To understand the relationship between myelination and neurofilament proteins in axons, we examined jimpy mutant mice with a point mutation in the proteolipid protein gene and dysmyelination in the central nervous system. The jimpy mice exhibited a characteristic similarity in neurofilament nature to the myelin-deficient mice in the peripheral nervous system reported previously. The following novel results were obtained in the jimpy mice: dysmyelinated axons, in which the amount of non-phosphorylated neurofilament-H was drastically increased without a significant reduction of the phosphorylated form, compared with the control myelinated axons, did not suffer any decrease in their diameters. Expression levels of all neurofilament subunit proteins and their mRNAs were enhanced in the central nervous system tissue. Because the above biochemical data were obtained from the cytoskeletal fraction, at least some of the increased neurofilament-H and -M proteins appeared to be coassembled into neurofilaments but remained non-phosphorylated. Axonal neurofilaments of the jimpy were, probably due to this abnormal stoichiometry and phosphorylation state in neurofilaments, more compact and random in alignment with less prominent cross-bridges than those of the control, providing possible evidence for disturbing the axonal transport of other organelles. These results suggest that myelination regulates both the expression and phosphorylation of neurofilament proteins, and is essential for the cytoplasmic organization of myelinated axons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1460-9568.1999.00820.x
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  • 7
    Electronic Resource
    Electronic Resource
    Radioautographic study on the obliteration of the ductus arteriosus botalli (1970)
    Springer
    Virchows Archiv 349 (1970), S. 10-20 
    Details
    ISSN: 1432-2307
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary From the findings obtained from the radioautographic study on the obliteration of the ductus arteriosus, the authors confirmed that the obliteration was not caused only by the proliferation of the cells in the intimal layer of the ductus, but rather by the migration of the cells into the lumen. When the embryos reached the final stage of fetal life, their ductus arteriosus possessed a specific property to obliterate after the blood flow ceased. Some discussions are devoted to the etiology of the patent ductus arteriosus.
    Notes: Zusammenfassung Durch autoradiographische Untersuchungen wurde gezeigt, daß die Obliteration des Ductus arteriosus Botalli, welche während einer kritischen Zeit abläuft, nicht nur durch die Wandzellen des Ductus sondern auch Elemente des strömenden Blutes getragen wird. Während der Phase der Proliferation besteht eine besondere Störanfälligkeit der Zellen, insbesondere gegenüber Sauerstoffmangel. Es scheint, daß die Suszeptibilität genisch verankert ist.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00548519
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  • 8
    Electronic Resource
    Electronic Resource
    Ultrastructures of glosso-palatal fusion after treatment of Meclozine-hydrochloride (1975)
    Springer
    Virchows Archiv 369 (1975), S. 7-17 
    Details
    ISSN: 1432-2307
    Keywords: Meclozine-hydrochloride ; Superficial Cells ; Degenerating Changes ; Glosso-Palatal Fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The behavior of the epithelial cells on the tongue and palatal processes during glosso-palatal fusion induced by Meclozine-hydrochloride was investigated with light and electron microscopes. Microscopical observations revealed that; at two or three days after the administration, the palatal processes were approximating to the tongue, and the superficial cells on the lateral sides of tongue became swollen and to have inclusion bodies, and in the epithelium on the medial sides of palatal processes, some inclusion bodies appeared too. At the time of contact, the superficial epithelial cells of tongue tended to degenerate markedly, and these degenerating cells were dissociated from the epithelium, and concurrently, superficial layer of the palatal epithelium became to have a lot of inclusion bodies. Between the two tissues, attachment devices developed just after the contact, and then, the cells in the intervening epithelial lining fell into a degeneration gradually. As well known, Meclozine-hydrochloride is one of important miner tranquilizers. From the findings mentioned above, it seemed to be possible that a heterotopic fusion caused with this drug is a result of a destruction of embryonic epithelium on tongue and palatal processes at a certain stage of development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00432457
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  • 9
    Electronic Resource
    Electronic Resource
    Carbon/Ceramics Composites with High Oxidation Resistance (1995)
    s.l. ; Stafa-Zurich, Switzerland
    Key engineering materials Vol. 108-110 (July 1995), p. 145-154 
    Details
    ISSN: 1013-9826
    Source: Scientific.Net: Materials Science & Technology / Trans Tech Publications Archiv 1984-2008
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Type of Medium: Electronic Resource
    URL: http://www.tib-hannover.de/fulltexts/2011/0528/01/44/transtech_doi~10.4028%252Fwww.scientific.net%252FKEM.108-110.145.pdf
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  • 10
    Electronic Resource
    Electronic Resource
    Synthesis of β-SiC by the Reaction of Gaseous SiO with Activated Carbon (1998)
    s.l. ; Stafa-Zurich, Switzerland
    Key engineering materials Vol. 159-160 (May 1998), p. 89-94 
    Details
    ISSN: 1013-9826
    Source: Scientific.Net: Materials Science & Technology / Trans Tech Publications Archiv 1984-2008
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Type of Medium: Electronic Resource
    URL: http://www.tib-hannover.de/fulltexts/2011/0528/01/44/transtech_doi~10.4028%252Fwww.scientific.net%252FKEM.159-160.89.pdf
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