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41.
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43.
P F Maness 《Journal of neuroscience research》1986,16(1):127-139
The advent of recombinant DNA technology has led to the identification in the DNA of normal animal cells of over 30 proto-oncogenes that are homologous to retroviral transforming genes. One of these encodes a protein kinase (pp60c-src) of unknown function, that is preferentially synthesized in brain and neural retina. Here the expression of pp60c-src in the peripheral nervous system was examined in sensory neurons from chick dorsal root ganglia with antisera raised against the transforming protein of Rous sarcoma virus (pp60v-src) expressed in Escherichia coli carrying the cloned v-src gene. This antiserum recognizes pp60c-src specifically in normal chicken cells. Western immunoblotting showed that dorsal root ganglia of stage 30 (day 6.5) chick embryos contained elevated levels of pp60c-src. Immunoperoxidase staining of neuron-enriched cultures prepared from chick dorsal root ganglia showed pp60c-src immunoreactivity in cells with neuronal morphology; flat, fibroblastic cells contained no detectable immunoreactivity. Indirect double immunofluorescence with pp60src antibodies and monoclonal antibodies against the 200-kD subunit of neurofilament protein confirmed that the cells expressing pp60c-src were neurons. Ninety-six percent of the neurofilament-positive cells were immunoreactive with pp60src antibodies, and conversely, all pp60c-src-positive cells were immunoreactive with neurofilament antibodies. pp60c-src immunofluorescence appeared to be distributed over the cell body, processes, and growth cones. These results clearly demonstrate that pp60c-src is a product of neurons and is expressed in sensory neurons in culture. 相似文献
44.
ALVARO MARTÍNEZ DEL POZO MARIA GASSET MERCEDES O
ADERRA JOS G. GAVILANES 《Chemical biology & drug design》1989,34(5):416-422
α-Sarcin binds one Zn(II) cation per protein molecule, with a Kd value of 0.9 mM, determined by equilibrium dialysis experiments. Ca(II), Mg(II), and Mn(II) do not bind to α-sarcin. Cd(II) and Co(II) also behave as Zn(II). The binding produces local modifications on the protein conformation affecting the microenvironment of tryptophan residues. The three cations modify the fluorescence emission of the protein. The near-u. v. circular dichroism spectrum of the protein is also altered. The binding of Zn(II) and related cations does not modify the secondary structure of the protein. The ribonucleolytic activity of a-sarcin is inhibited upon Zn(II) binding, but no alteration of the ability of the protein to aggregate phospholipid vesicles has been observed. 相似文献
45.
目的探讨内源性热休克蛋白90(HSP90)在缺氧心肌细胞丝氨酸苏氨酸蛋白激酶(AKT)相关信号通路中的作用。方法建立新生Wistar大鼠心肌细胞缺氧模型,将细胞分为正常组、缺氧组、加入HSP90特异性阻断剂格尔德霉素后再缺氧组(格尔德霉素+缺氧组)。于缺氧后1、3、6、12、24、48h用噻唑蓝法检测心肌细胞的活力;缺氧24h,原位缺口末端标记法检测心肌细胞凋亡指数(AI);缺氧1、3、6、12、24h,蛋白质印迹法检测大鼠心肌细胞中内源性HSP90及AKT表达水平。结果(1)缺氧24、48h,缺氧组、格尔德霉素+缺氧组细胞活力均较正常组明显下降(P〈0.05);格尔德霉素+缺氧组细胞活力缺氧12h即开始明显下降,缺氧48h时明显低于缺氧组(P〈0.05)。(2)缺氧24h,缺氧组细胞AI为(10.7±1.2)%,明显高于正常组[(1.9±0.3)%.P〈0.05];格尔德霉素+缺氧组细胞AI为(26、3±5.3)%,明显高于缺氧组(P〈0.01)。(3)缺氧12h,缺氧组心肌细胞内源性HSP90及AKT表达水平高于正常组与格尔德霉素+缺氧组;缺氧24h,缺氧组有所下降.格尔德霉素+缺氧组则下降更明显。结论内源性HSP90对维持心肌细胞的活力有重要作用.缺氧心肌细胞AKT表达水平可受内源性HSP90表达水平的影响。 相似文献
46.
目的观察葛根素对血管性痴呆(VaD)患者认知功能和听觉事件相关电位P300的影响。方法将70例VaD患者随机分成两组各35例,葛根素治疗组和对照组。应用简易精神状态检查量表(MMSE)评定两组患者治疗前后认知功能状况,并进行治疗前后P300检查。同时记录药物不良反应。结果两组各35例进入结果分析。治疗前两组MMSE评分、P300的潜伏期及波幅差异无显著性(均P〉0.05)。治疗14d、30d时,两组MMSE评分显著提高(均P〈0.01),P300潜伏期均有缩短,波幅均有提高(葛根素组P〈0.01,对照组P〈0.05);治疗后14d时认知功能改善葛根素组明显优于对照组(总有效率分别为91.4%,71.4%)(P〈0.05)。两组治疗期间无严重不良反应。结论葛根素能够改善VaD患者的认知功能,这可能与葛根素的扩血管、脑保护作用有关。 相似文献
47.
Terry Joe Sprinkle Julia F. Agee Russell B. Tippins C. Richard Chamberlain Guy B. Faguet George H. DeVries 《Brain research》1987,426(2):349-357
Monoclonal antibodies against human and bovine 2′:3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) were generated by fusing FOX-NY myeloma cells with spleen cells from RBF/Dn mice previously immunized with the purified brain antigens. The enzyme isolated from bovine brain was quite basic, with an isoelectric point of 9.71 and both the bovine and human enzymes consisted of a closely spaced doublet at approximately 44 and 46 kDa on SDS-PAGE. Six monoclonals were identified as strongly recognizing the enzyme on both ELISA plates and on immunoblots of whole brain protein. Four monoclonals very weakly cross-reacted with guinea pig myelin basic protein. In contrast with two previous reports, some of our monoclonal antibodies did immunostain 2 or 3 protein bands in peripheral nerve, two bands closely corresponding to those immunostained in central nervous system (CNS) myelin, the Wolfgram protein fraction and in acetone powders of whole brain. Each of the 6 monoclonals reacting strongly on immunoblots recognized the enzyme in from 2 to 5 of the species examined (human, bovine, rat, mouse and rabbit). In addition, all 6 monoclonals that immunostained the enzyme in whole brain, myelin and Wolfgram protein immunoblots recognized both CNP1 (44 kDa) and CNP2 (46 kDa). The two closely spaced protein bands observed on SDS-PAGE and previously stained on immunoblots of CNS CNPase using polyvalent rabbit anti-bovine CNPase antisera, and now different monoclonal antibodies, appear to be immunologically related and to contain highly conserved sequences. 相似文献
48.
49.
Kazuyoshi Kataoka Katsuo Furukawa Kohichi Nagao Nobuhisa Ishii Hiromichi Tsuru 《International journal of urology》2007,14(8):764-768
AIM: To investigate the participation of adenosine receptors in the adenosine 5'-triphosphate (ATP)-induced relaxation in the corpus cavernosum penis (CCP) of rabbits. METHODS: The ATP-induced relaxation was assessed on the noradrenaline precontracted CCP of rabbits in the presence and absence of 8-(3-chlorostyryl)caffeine (CSC); an adenosine A(2A) receptor antagonist; alloxazine and MRS1754; adenosine A(2B) receptor antagonists; and ARL67156, an inhibitor of ecto-nucleoside triphosphate diphosphohydrolases. RESULTS: Adenosine and ATP relaxed the noradrenaline precontracted CCP of rabbits in a concentration-dependent manner. The adenosine- and ATP-induced relaxations were suppressed by alloxazine and MRS1754, but not by 8-(3-chlorostyryl)caffeine. ARL67156 potentiated the ATP-induced relaxation but not the adenosine-induced one. MRS1754 suppressed the ATP-induced relaxation potentiated by ARL67156. CONCLUSIONS: The above results suggest that, in the CCP of rabbits, the adenosine receptor mediating adenosine-induced relaxation is of the A(2B) receptor and the ATP directly causes relaxation through the A(2B) receptor on the CCP. 相似文献
50.
The present study was performed in order to follow the response of astroglial cells in the rat hippocampus to chronic low-level lead exposure. The experiments combined immunohistochemistry using anti-glial fibrillary acidic protein (GFAP) antibody and conventional transmission electron microscopy (EM). Chronic administration with drinking water [1 g% w/v (subclinical dose) of lead acetate dissolved in distilled water] was started through the mother's milk when pups were 7 days old. Following weaning, experimental offspring were treated for 3 months with the same concentration of adulterated water. The group of intoxicated animals and their controls were sacrificed by perfusion-fixation at 30, 60, and 90 days of exposure. After 60 days of lead treatment, staining of GFAP-positive cells demonstrated an astroglial transformation from the quiescent to the reactive state, characterized by an increase in GFAP. In control rats no changes in GFAP immunostaining were observed. The intensity of the astroglial response was enhanced after 90 days of lead intoxication, showing an increment of GFAP immunoreactivity. Quantification of these changes was made by computerized image analysis, confirming that the sectional areas of the astroglia in lead-exposed animals were larger than those in controls. These results are consistent with the ultrastructural alterations. Simultaneously with the increment in gliofilaments, intranuclear inclusions were seen in some astrocytes. The mechanisms by which lead affects astrocytes are unknown. Probably the astroglial changes induced by lead intoxication produce microenvironmental modifications that may disturb the neuronal function. 相似文献