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Heterogeneity in the Expression Pattern of Two Ganglioside Synthase Genes During Mouse Brain Development 总被引:5,自引:1,他引:5
†Akihito Yamamoto †Masashi Haraguchi †Shuji Yamashiro †‡Satoshi Fukumoto †Keiko Furukawa †Kogo Takamiya Mitsuru Atsuta †§Hiroshi Shiku † Koichi Furukawa 《Journal of neurochemistry》1996,66(1):26-34
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Maiko Miyata Masatoshi Ichihara Orie Tajima Sayaka Sobue Mariko Kambe Kazumitsu Sugiura Koichi Furukawa Keiko Furukawa 《Biochemical and biophysical research communications》2014
Although expression of gangliosides and their synthetic enzyme genes in malignant melanomas has been well studied, that in normal melanocytes has been scarcely analyzed. In particular, changes in expression levels of glycosyltransferase genes responsible for ganglioside synthesis during evolution of melanomas from melanocytes are very important to understand roles of gangliosides in melanomas. Here, expression of glycosyltransferase genes related to the ganglioside synthesis was analyzed using RNAs from cultured melanocytes and melanoma cell lines. Quantitative RT-PCR revealed that melanomas expressed high levels of mRNA of GD3 synthase and GM2/GD2 synthase genes and low levels of GM1/GD1b synthase genes compared with melanocytes. As a representative exogenous stimulation, effects of ultraviolet B (UVB) on the expression levels of 3 major ganglioside synthase genes in melanocytes were analyzed. Although direct UVB irradiation of melanocytes caused no marked changes, culture supernatants of UVB-irradiated keratinocytes (HaCaT cells) induced definite up-regulation of GD3 synthase and GM2/GD2 synthase genes. Detailed examination of the supernatants revealed that inflammatory cytokines such as TNFα and IL-6 enhanced GD3 synthase gene expression. These results suggest that inflammatory cytokines secreted from UVB-irradiated keratinocytes induced melanoma-associated ganglioside synthase genes, proposing roles of skin microenvironment in the promotion of melanoma-like ganglioside profiles in melanocytes. 相似文献
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Fukumoto Satoshi; Yamamoto Akihito; Hasegawa Tomokazu; Abe Kuniko; Takamiya Kogo; Okada Masahiko; Min Zhao Jin; Furukawa Keiko; Miyazaki Hiroshi; Tsuji Yoshiro; Goto George; Suzuki Misao; Shiku Hiroshi; Furukawa Koichi 《Glycobiology》1997,7(8):1111-1120
Several lines of transgenic mice with gangliosides GM2/GD2 synthasegene were established, and the expression levels of the transgenein brain, liver, spleen and thymus were analyzed by comparingwith those in their litter mates. Among four tissues, brainand skin showed markedly high expression levels of the transgenein Northern blotting. Particularly, transgenic mice skin showedabout 10-fold higher expression of GM2/GD2 synthase gene thanthe wild type mice skin. Therefore, alterations in the morphology,glycolipid components, and responses to the exogenous stimulationsin the transgenic mice skin were examined. Gangliosides in thetransgenic skin were dramatically converted from GM3 to GM1,whereas no morphological changes were observed. However, whenskin flap test was performed with insertion of nylon membranesunder the skin flaps, much stronger inflammatory reactions consistingof edema, marked thickness, and cell infiltration were observedin the transgenic mice compared with the wild type. Similarenhanced inflammatory reaction was also observed in the skininjected by silicon gel, and in the peritoneal reaction to theinjected casein. Main cell population in these inflammatoryreactions consisted of neutrophils, suggesting an increasedsensitivity of neutrophils to chemo-tactic factors in the transgenicmice. ganglioside glycosyltransferase GM2 GD2 synthase skin transgenic mouse 相似文献
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Regional distribution of brain-derived neurotrophic factor mRNA in the adult mouse brain. 总被引:50,自引:4,他引:46
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Brain-derived neurotrophic factor (BDNF) is a protein that allows the survival of specific neuronal populations. This study reports on the distribution of the BDNF mRNA in the adult mouse brain, where the BDNF gene is strongly expressed, using quantitative Northern blot analysis and in situ hybridization. All brain regions examined were found to contain substantial amounts of BDNF mRNA, the highest levels being found in the hippocampus followed by the cerebral cortex. In the hippocampus, which is also the site of highest nerve growth factor (NGF) gene expression in the central nervous system (CNS), there is approximately 50-fold more BDNF mRNA than NGF mRNA. In other brain regions, such as the granule cell layer of the cerebellum, the differences between the levels of BDNF and NGF mRNAs are even more pronounced. The BDNF mRNA was localized by in situ hybridization in hippocampal neurons (pyramidal and granule cells). These data suggest that BDNF may play an important role in the CNS for a wide variety of adult neurons. 相似文献
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Nobuyuki Kurosawa Yukiko Yoshida Naoya Kojima Shuichi Tsuji 《Journal of neurochemistry》1997,69(2):494-503
Abstract: A comparative study was undertaken to correlate the immunohistochemical localization of polysialic acid (PSA) and the in situ localization of ST8Sia II mRNA. In situ hybridization of postnatal day 3 mouse brain showed high levels of ST8Sia II mRNA expression in the cerebral neocortex, striatum, hippocampus, subiculum, medial habenular nucleus, thalamus, pontine nuclei, and inferior colliculus; intermediate-level expression in the olfactory bulb, hypothalamus, superior colliculus, and cerebellum; and low-level expression in other regions. The distribution of ST8Sia II mRNA in the neocortex and cerebellum coincided with the immunohistochemical localization of PSA. During brain development, ST8Sia II mRNA started decreasing and had almost disappeared by postnatal day 14. Comparison between ST8Sia II and IV mRNA expression was also undertaken by northern blot analysis and competitive PCR analysis. During the late embryonic to early postnatal stages of the mouse CNS, the ST8Sia II mRNA showed abundant mRNA expression compared with the ST8Sia IV mRNA. Competitive PCR analysis of the adult mouse CNS showed weak expression of the two genes in the olfactory bulb, thalamus, hippocampus, and eyes. The regional and transient expression of ST8Sia II mRNA coincides with that of PSA, suggesting that ST8Sia II is closely involved in the biosynthesis and expression of PSA in the developing mouse CNS. 相似文献
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Henion TR Zhou D Wolfer DP Jungalwala FB Hennet T 《The Journal of biological chemistry》2001,276(32):30261-30269
The distinction between the different classes of glycolipids is conditioned by the action of specific core transferases. The entry point for lacto-series glycolipids is catalyzed by the beta1,3 N-acetylglucosaminyltransferase GlcNAc(beta1,3)Gal(beta1,4)Glc-ceramide (Lc3) synthase enzyme. The Lc3 synthase activity has been shown to be regulated during development, especially during brain morphogenesis. Here, we report the molecular cloning of a mouse gene encoding an Lc3 synthase enzyme. The mouse cDNA included an open reading frame of 1131 base pairs encoding a protein of 376 amino acids. The Lc3 synthase protein shared several structural motifs previously identified in the members of the beta1,3 glycosyltransferase superfamily. The Lc3 synthase enzyme efficiently utilized the lactosyl ceramide glycolipid acceptor. The identity of the reaction products of Lc3 synthase-transfected CHOP2/1 cells was confirmed by thin-layer chromatography immunostaining using antibodies TE-8 and 1B2 that recognize Lc3 and Gal(beta1,4)GlcNAc(beta1,3)Gal(beta1,4)Glc-ceramide (nLc4) structures, respectively. In addition to the initiating activity for lacto-chain synthesis, the Lc3 synthase could extend the terminal N-acetyllactosamine unit of nLc4 and also had a broad specificity for gangliosides GA1, GM1, and GD1b to generate neolacto-ganglio hybrid structures. The mouse Lc3 synthase gene was mainly expressed during embryonic development. In situ hybridization analysis revealed that that the Lc3 synthase was expressed in most tissues at embryonic day 11 with elevated expression in the developing central nervous system. Postnatally, the expression was restricted to splenic B-cells, the placenta, and cerebellar Purkinje cells where it colocalized with HNK-1 reactivity. These data support a key role for the Lc3 synthase in regulating neolacto-series glycolipid synthesis during embryonic development. 相似文献
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Wang L Takaku S Wang P Hu D Hyuga S Sato T Yamagata S Yamagata T 《Glycoconjugate journal》2006,23(5-6):303-315
GD1a was previously shown responsible for regulating cell motility, cellular adhesiveness to vitronectin, phosphorylation
of c-Met and metastatic ability of mouse FBJ osteosarcoma cells. To determine the particular molecules regulated by GD1a,
FBJ cells were assessed for tumor-related gene expression by semi-quantitative RT-PCR. Caveolin-1 and stromal interaction
molecule 1 (Stim1) expression in FBJ-S1 cells, rich in GD1a, were found to be 6 and 4 times as much, respectively, than in
FBJ-LL cells devoid of GD1a. Enhanced production of caveolin-1 in protein was confirmed by Western blotting. A low-metastatic
FBJ-LL cell variant, having high GD1a expression through β1-4GalNAcT-1 (GM2/GD2 synthase) cDNA transfection (Hyuga S, et al, Int J Cancer 83: 685-91, 1999), showed enhanced production of caveolin-1 and Stim1 in mRNA and protein, compared to mock-transfectant
M5. Incubation of FBJ-M5 cells with exogenous GD1a augmented the expression of caveolin-1 in mRNA and protein and Stim1 in
mRNA as well. Treatment of FBJ-S1 with fumonisin B1, an inhibitor of N-acylsphinganine synthesis, for 15 days caused the complete depletion of gangliosides and suppressed the expression of caveolin-1
and Stim1. St3gal5 siRNA transfected cells showed decreased expression of caveolin-1 and Stim1 mRNA, as well as St3gal5 mRNA.
These findings clearly indicate ganglioside GD1a to be involved in the regulation of the transformation suppressor genes,
caveolin-1 and Stim1. Moreover, treatment with GD1a of mouse melanoma B16 cells and human hepatoma HepG2 cells brought about
elevated expression of caveolin-1 and Stim1.
Li Wang and Shizuka Takaku are equal contributors to the present work 相似文献
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Fernando Kreutz Fernanda dos Santos Petry Melissa Camassola Vanessa Schein Fátima C.R. Guma Nance Beyer Nardi Vera Maria Treis Trindade 《Gene》2013
Mucopolysaccharidosis I (MPS I) is a congenital disorder caused by the deficiency of α-l-iduronidase (IDUA), with the accumulation of glycosaminoglycans (GAGs) in the CNS. Although GAG toxicity is not fully understood, previous works suggest a GAG-induced alteration in neuronal membrane composition. This study is aimed to evaluate the levels and distribution of gangliosides and cholesterol in different brain regions (cortex, cerebellum, hippocampus and hypothalamus) in a model using IDUA knockout (KO) mice (C57BL/6). Lipids were extracted with chloroform–methanol and then total gangliosides and cholesterol were determined, followed by ganglioside profile analyses. While no changes in cholesterol content were observed, the results showed a tissue dependent ganglioside alteration in KO mice: a total ganglioside increase in cortex and cerebellum, and a selective presence of GM3, GM2 and GD3 gangliosides in the hippocampus and hypothalamus. To elucidate this, we evaluated gene expression of ganglioside synthesis (GM3, GD3 and GM2/GD2 synthases) and degradation of (Neuraminidase1) enzymes in the cerebellum and hippocampus by RT-sq-PCR. The results obtained with KO mice showed a reduced expression of GD3 and GM2/GD2 synthases and Neuraminidase1 in cerebellum; and a decrease in GM2/GD2 synthase and Neuraminidase1 in the hippocampus. These data suggest that the observed ganglioside changes result from a combined effect of GAGs on ganglioside biosynthesis and degradation. 相似文献
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目的研究adam10基因在成年小鼠中枢神经系统表达的脑区分布特点以及细胞类型。方法构建小鼠源性adam10 cRNA探针,通过原位杂交技术,观察adam10 mRNA在成年小鼠中枢神经系统分布特点,并在原位杂交后进行免疫组织化学染色,把adam10原位杂交信号和神经元、星形胶质细胞特异性细胞标记物进行双标,观察adam10基因表达的细胞类型。结果 Adam10基因在成年小鼠大脑皮层、海马、丘脑和小脑中表达,原位杂交后进行免疫组织化学染色结果显示adam10原位杂交阳性信号主要和神经元标记物NeuN共标,而和星形胶质细胞标记物GFAP不共标。结论本研究证实了在成年小鼠中枢神经系统中adam10基因在大脑皮层、海马、丘脑和小脑中都有表达;并且首次明确了大脑中ad-am10基因主要在神经元中表达,在星形胶质细胞中不表达,小脑中主要在小脑颗粒细胞和蒲肯野细胞中表达。 相似文献
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Induction of Src-suppressed C kinase substrate (SSeCKS) in vascular endothelial cells by bacterial lipopolysaccharide. 总被引:4,自引:0,他引:4
Hiroshi Kitamura Keisuke Okita Daisuke Fujikura Koshi Mori Toshihiko Iwanaga Masayuki Saito 《The journal of histochemistry and cytochemistry》2002,50(2):245-255
We isolated cDNA of the mouse homologue of the src-suppressed C kinase substrate (SSeCKS) and analyzed the effects of lipopolysaccharide (LPS) injection on the tissue expression pattern of this protein. Northern blotting analysis showed that SSeCKS mRNA was expressed abundantly in the testis but at undetectable levels in other tissues of untreated control mice. Intraperitoneal administration of LPS strongly induced SSeCKS mRNA expression in the lung, heart, liver, spleen, kidney, lymph node, adrenal gland, and pituitary gland, as well as in the brain. In lung and spleen, the SSeCKS mRNA levels increased almost 10-fold at 1 hr after LPS injection and persisted at high levels until 4 hr. Both in situ hybridization and immunohistochemical studies revealed that LPS administration conspicuously elevated expression of SSeCKS mRNA and protein in vascular endothelial cells of several organs. Ectopic expression of SSeCKS caused loss of cytoplasmic F-actin fibers in the mouse endothelial cell line LEII. These results indicate that SSeCKS is one of the major LPS-responsive proteins and may participate in alteration of cytoskeletal architecture in endothelial cells during inflammation. 相似文献
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Gene Expression of the Neuropeptide-Processing Enzyme Carboxypeptidase E in Rat Photoreceptor Cells 总被引:1,自引:1,他引:0
Robert W. Nickells† Cassandra L. Schlamp Alexandra C. Newton† David S. Williams 《Journal of neurochemistry》1993,61(5):1891-1900
Abstract: We characterized the spatial expression of mRNA for the enzyme Carboxypeptidase E (CPE) in the Long-Evans rat retina. CPE is involved in the processing of neuroactive peptides to a mature form. A cDNA encoding the 3' terminus of CPE mRNA was cloned by polymer-ase chain reaction amplification of rat retina single-stranded cDNA. The sequence of this cDNA was identical to a rat genomic clone for CPE and nearly identical (130/ 132 nucleotides) to a cDNA for rat brain CPE. In addition, the cDNA hybridized to a single allele on Southern blots and to a 2.1-kb mRNA on northern blots of both rat brain and retina. These data support the conclusion of others that CPE is a single-copy gene in the rat. In cell fractionation experiments, the majority of CPE mRNA fractionated with rod opsin mRNA, suggesting that CPE is expressed predominantly in rod photoreceptors. The high abundance of CPE mRNA in photoreceptors was confirmed by in situ hybridization studies, although CPE was detected at lower levels in other retinal cell types as well. The presence of abundant levels of the mRNA of a neuro-peptide-processing enzyme in photoreceptor cells suggests that photoreceptors may utilize neuropeptides for normal function. 相似文献
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James A. Waschek Julie Ellison Dawn T. Bravo Vance Handley 《Journal of neurochemistry》1996,66(4):1762-1765
Abstract: Vasoactive intestinal peptide (VIP) exhibits pronounced effects on the growth rate of cultured mouse embryonic day (E) 9.5 embryos and acts in tissue culture as a potent glial mitogen and neuron survival factor. However, previous studies using immunohistochemistry or in situ hybridization in the rat have not revealed the presence and location of VIP or VIP mRNA in the early developing embryo CNS. Using a sensitive in situ hybridization assay with a 33 P-labeled riboprobe, we show here that the VIP gene is expressed at least as early as E11 in the mouse hindbrain. Northern blot analysis on RNA from brain dissected from mouse embryos beginning at E14 confirmed that a correct-size mRNA for VIP was present by E14 and at later time points. Expression of the VIP2 receptor gene was also detected by northern analysis in E14 mouse brains. These studies support the hypothesis that VIP produced by the embryo exerts important effects on embryonic nervous system development. 相似文献