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Studies on the reproductive and developmental biology of Cichlasoma dimerus (Percifomes, Cichlidae). 二角鳞鱼生殖发育生物学的研究(鲈形目,姬鱼科)。
M. Pandolfi, M. Cánepa, F. Meijide, Felipe Alonso, G. R. Vázquez, M. Maggese, P. Vissio
Many characteristics of the South American teleost fish Cichlasoma dimerus (body size, easy breeding, undemanding maintenance) make it amenable to laboratory studies. In the last years, many of the fundamental aspects of its reproductive and developmental biology have been addressed in our laboratory. Rather recently, the immunohistochemical localization of pituitary hormones involved in reproduction and in background color adaptation has been described in both adult and developing individuals, and the role of FSH in ovarian differentiation has been established. These findings have been correlated with mapping of some of their brain-derived controlling hormones. The latter include brain-derived gonadotropins which were shown to be active in vitro in the control of pituitary hormone secretions. The emerging picture shows C. dimerus as an interesting species in which many of their basic features have already been investigated and which conform a solid platform for comparative studies correlating neurohormones, pituitary hormones and behavior, from the molecular to the organismic level.
南美洲硬骨鱼的许多特点(体型大,容易繁殖,不需要维护)使其适合实验室研究。在过去的几年中,其生殖和发育生物学的许多基本方面已经在我们的实验室得到解决。最近,在成人和发育个体中,参与生殖和背景色适应的垂体激素的免疫组织化学定位已被描述,并且FSH在卵巢分化中的作用已被确立。这些发现与他们脑源性控制激素的绘制有关。后者包括脑源性促性腺激素,它在体外被证明在控制垂体激素分泌方面是活跃的。新出现的图片显示,C. dimerus是一个有趣的物种,它们的许多基本特征已经被研究过,并且为从分子到有机体水平的神经激素、垂体激素和行为的比较研究提供了坚实的平台。
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引用次数: 51
Influence of 2-methoxyestradiol on MCF-7 cells: an improved differential interference contrasting technique and Bcl-2 and Bax protein expression levels. 2-甲氧基雌二醇对MCF-7细胞的影响:一种改进的差异干扰对比技术和Bcl-2和Bax蛋白表达水平
A. Joubert, S. Marais, C. Maritz
Proteins of the B-cell lymphoma 2 family are crucial for the regulation of apoptosis. B-cell lymphoma 2-associated X is a pro-apoptotic protein, while B-cell lymphoma 2 protein opposes apoptosis. The influence of 1 microM 2-methoxyestradiol was investigated on the expression levels of these two proteins in MCF-7 cells. 2-Methoxyestradiol exposure did not influence B-cell lymphoma 2 protein expression levels after 24 h of exposure. In contrast, B-cell lymphoma 2-associated X protein levels were significantly reduced. An improved differential interference contrasting technique revealed compromised cell density and the presence of a mitotic block in exposed cells. The study proposes that the influence of 2-methoxyestradiol on the expression of these proteins may be time- and cell type dependent and thus not evident during the mitotic block observed. Investigation of the regulation of the B-cell lymphoma 2 family will allow researchers to consider signaling pathways for diseases where apoptosis can potentially be controlled.
b细胞淋巴瘤2家族蛋白对细胞凋亡的调控至关重要。b细胞淋巴瘤2相关蛋白X是促凋亡蛋白,而b细胞淋巴瘤2蛋白则是抗凋亡蛋白。研究了1 μ m 2-甲氧基雌二醇对MCF-7细胞中这两种蛋白表达水平的影响。暴露24小时后,2-甲氧基雌二醇不影响b细胞淋巴瘤2蛋白表达水平。相比之下,b细胞淋巴瘤2相关的X蛋白水平显著降低。一种改进的微分干涉对比技术揭示了暴露细胞中受损的细胞密度和有丝分裂阻滞的存在。该研究提出,2-甲氧基雌二醇对这些蛋白表达的影响可能是时间和细胞类型依赖的,因此在观察到的有丝分裂阻断期间不明显。对b细胞淋巴瘤2家族调控的研究将使研究人员能够考虑细胞凋亡可能被控制的疾病的信号通路。
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引用次数: 14
A 66-kDa protein of bovine hypophyseal Pars tuberalis induces luteinizing hormone release from rat Pars distalis. 牛垂体结节部的66-kDa蛋白诱导大鼠远端部释放黄体生成素。
M. Lafarque, L. Oliveros
In this study, evidence for a factor secreted by bovine hypophyseal pars tuberalis that stimulates luteinizing hormone (LH) release from rat pars distalis cells is shown. The secretion products of bovine pars tuberalis cells into the culture medium were assayed on dispersed rat pars distalis cells in 30 min incubations and superfusion experiments. The culture medium from pars tuberalis total cell populations, added at a dose of 6 microg per tube, induced the greater LH release from pars distalis cells, without effect on follicle stimulating hormone (FSH) release. After pars tuberalis cells separation on a discontinuos Percoll gradient, only the culture medium of cells from 50 and 60% strength Percoll were able to release LH from rat pars distalis cells. Therefore, cell fractions from 50 and 60% strenght Percoll were cultured together. To elicit maximal LH release (6 times the basal output), with the addition of 2 microg of pars tuberalis protein was required, suggesting that these cells produce the factor or factors which affect pars distalis gonadotrope cells. After applying the pars tuberalis culture medium on 12% SDS-PAGE, the band with biological activity was that of 66-kDal. Fifty ng protein of its eluate released almost 9 times the basal output of LH from pars distalis cells. Results suggest a modulating effect of a protein from the bovine pars tuberalis on rat cultured gonadotrope cells from the pars distalis.
在这项研究中,证据表明牛垂体结节部分泌的一种因子刺激大鼠远端部细胞释放黄体生成素(LH)。在分散的大鼠远部细胞上进行30min的培养和灌注实验,检测牛结节部细胞分泌到培养基中的产物。从结节部总细胞群中提取的培养基,每管添加6 μ g,诱导远端部细胞释放更多的LH,但对促卵泡激素(FSH)的释放没有影响。在不连续Percoll梯度下分离大鼠结节部细胞后,只有50%和60% Percoll强度的培养液才能释放出大鼠远部细胞中的LH。因此,将强度为50%和60%的Percoll细胞组分一起培养。为了获得最大的LH释放量(6倍于基础输出量),需要添加2毫克的结节部蛋白,这表明这些细胞产生影响远部促性腺激素细胞的因子。在12% SDS-PAGE上涂布结核菌培养基后,具有生物活性的条带为66-kDal。其洗脱物的50 ng蛋白释放的LH几乎是远部细胞基础输出量的9倍。结果表明,牛结节部蛋白对培养的大鼠远部促性腺激素细胞有调节作用。
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引用次数: 1
Isolation of epithelial cells, villi and crypts from small intestine of pigeons (Columba livia). 鸽子小肠上皮细胞、绒毛和隐窝的分离。
Oscar Mac Donal, J. G. Chediack, E. Caviedes-Vidal
The isolation of viable enterocytes, villi and crypts from the small intestine of a feral bird (Columba livia) is important for performing physiological experiments in ecologically relevant processes of membrane transport. The effectiveness of mechanical disruption, enzymatic digestion and chelating agents were compared. The objectives were to isolate enterocytes, villi and crypts from the small intestine of young pigeons; to evaluate the viability of the isolated intestinal epithelial cells isolated; and to verify the integrity of enterocytes by biochemical features. Enzymatic and mechanical methods yielded both elongated columnar and spherical cells. With the chelating method villi and crypts were obtained. All methods produced a high yield of intestinal epithelial cells with about 50% viability. Brush border enzymes (sucrase-isomaltase and alkaline phosphatase) activities were high and, as reported in chickens, they did not differ along the intestinal villus-crypt axis. Although the three methods have good viabilities, the enzymatic technique gives the best yield in cell number, while the chelating method provides the highest populations of morphologically distinctive villi and crypts.
从野鸟(Columba livia)的小肠中分离出活的肠细胞、绒毛和隐窝,对于在膜运输的生态相关过程中进行生理实验具有重要意义。比较了机械破坏剂、酶消化剂和螯合剂的效果。目的是从雏鸽的小肠中分离肠细胞、绒毛和隐窝;评价分离的肠上皮细胞的活力;并通过生化特征验证肠细胞的完整性。酶和机械方法都产生了细长的柱状细胞和球形细胞。用螯合法获得绒毛和隐窝。所有方法均可获得高产量的肠上皮细胞,存活率约为50%。刷状边界酶(蔗糖酶-异麦芽糖酶和碱性磷酸酶)活性高,正如在鸡中报道的那样,它们在肠绒毛-隐窝轴上没有差异。虽然这三种方法都具有良好的可行性,但酶法在细胞数量上的产量最高,而螯合法在形态上独特的绒毛和隐窝的数量最多。
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引用次数: 17
Isolation and molecular characterization of a cax gene from Capsella bursa-pastoris. 法氏囊荠菜cax基因的分离及分子特征分析。
Juan Lin, Wen Zhang, Mingzhu Shi, Xinglong Wang, Xiaofen Sun, K. Tang
A new cation exchangers (CAXs) gene was cloned and characterized from Capsella bursa-pastoris by rapid amplification of cDNA ends (RACE). The full-length cDNA sequence of cax from C. bursa-pastoris (designated as Cbcax51) was 1754 bp containing a 1398 bp open reading frame encoding a polypeptide of 466 amino-acid residues with a calculated molecular mass of 50.5 kDa and an isoelectric point of 5.69. The predicted CbCAX51 contained an IMP dehydrogenase/GMP reductase domain, two Na+/Ca2+ exchanger protein domains. Comparative and bioinformatics analyses revealed that CbCAX51 showed extensive homology with CAX from other plant species. The expression analysis by different treatments indicated that Cbcax51 could be activated by cold triggering and was related to the cold acclimation process, but its expression is regulated negatively by drought and not affected by ABA or salt.
从法氏囊荠菜(Capsella bursa-pastoris)中克隆了一个新的阳离子交换器(CAXs)基因,并利用cDNA末端快速扩增技术(RACE)对其进行了鉴定。法氏囊酵母(C. bursa-pastoris) cax全长cDNA序列为1754 bp,包含一个1398 bp的开放阅读框,编码466个氨基酸残基的多肽,计算分子量为50.5 kDa,等电点为5.69。预测的CbCAX51含有一个IMP脱氢酶/GMP还原酶结构域,两个Na+/Ca2+交换蛋白结构域。比较和生物信息学分析表明,CbCAX51与其他植物物种的CAX具有广泛的同源性。不同处理的表达分析表明,Cbcax51可被冷触发激活,与冷驯化过程有关,但其表达受干旱负调控,不受ABA和盐的影响。
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引用次数: 6
Heme metabolism enzymes are dynamically expressed during Xenopus embryonic development. 非洲爪蟾胚胎发育过程中血红素代谢酶的动态表达。
Jianli Shi, W. Mei, Jing Yang
As the key component of many hemoproteins (heme-containing proteins), heme is involved in a broad range of biological processes. Enzymes required for heme biosynthesis and degradation pathways are evolutionarily conserved. While heme metabolism has been studied extensively, the expression of heme metabolism enzymes during development has not been described. Here, we report that all heme biosynthases and two heme oxygenases, which initiate heme degradation, are dynamically expressed during Xenopus embryonic development. All heme synthases, with the exception of aminolevulinic acid synthase 2, are maternally expressed. At neurula stage, heme synthases are expressed in the developing neural tissue and in migrating neural crest cells. At the swimming tadpole stage, expression of heme synthases can be detected in multiple lineages, including eyes, neural crest cells, developing central nervous system, ventral blood island, pronephron, and pronephric tubule. Similar to heme synthases, heme oxygenases are expressed maternally. Zygotic expression of heme oxygenases is mainly restricted to the developing neural and neural crest lineages. Unlike heme synthases, heme oxygenases are not expressed in the ventral blood island and are expressed at a very low level in the pronephron and pronephric tubule. This indicates that heme metabolism may play important roles during development.
作为许多血红蛋白(含血红素蛋白)的关键成分,血红素参与了广泛的生物过程。血红素生物合成和降解途径所需的酶是进化保守的。虽然血红素代谢已被广泛研究,但血红素代谢酶在发育过程中的表达尚未被描述。在这里,我们报道了所有血红素生物合成酶和两种血红素加氧酶,启动血红素降解,在爪蟾胚胎发育过程中动态表达。除了氨基乙酰丙酸合成酶2外,所有的血红素合成酶都是母系表达的。在神经发育期,血红素合成酶在发育中的神经组织和迁移的神经嵴细胞中表达。在游泳蝌蚪阶段,血红素合成酶的表达可以在多个谱系中检测到,包括眼睛、神经嵴细胞、发育中的中枢神经系统、腹侧血岛、肾原细胞和肾原小管。与血红素合成酶类似,血红素加氧酶由母体表达。血红素加氧酶的合子表达主要局限于发育中的神经和神经嵴谱系。与血红素合成酶不同,血红素加氧酶在腹侧血岛中不表达,在肾原素和肾原小管中表达水平很低。这表明血红素代谢可能在发育过程中起重要作用。
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引用次数: 13
Sequential expressions of Notch1, Jagged2 and Math1 in molar tooth germ of mouse. Notch1、Jagged2和Math1在小鼠磨牙胚中的序列表达。
S. Borkosky, H. Nagatsuka, Y. Orita, H. Tsujigiwa, Junko Yoshinobu, M. Gunduz, A. Rodríguez, L. Missana, K. Nishizaki, N. Nagai
The Notch signaling pathway is an evolutionary conserved mechanism that plays an important role in cell-cell communication and cell fate in a wide range of tissues. The mammalian family of Notch receptors consists of 4 members: Notch1/2/3/4. The Notch ligand family consists of 5 members: Delta1/3/4 and Jagged1/2. Math1 encodes a murine basic helix-loop-helix (bHLH) transcription factor that acts as positive regulator of cell differentiation. Recently, links between Notch and Math1 pathways were demonstrated in various tissues. Expression of Notch1, Jagged2 and Math1 were analyzed in the mouse molar tooth germ during embryonic stage (E) 13 and E15 and during postnatal stage (PN) 1, PN3, PN5, PN10 and PN14 by using in situ hybridization. Positive Notch1 expression was found at the tooth bud during embryonic stages, but its expression was absent from the basal cells in contact with the dental mesenchyme. Jagged2 and Math1 were strongly expressed in differentiated ameloblasts and odontoblasts and Math1 strong expression was even maintained until PN14 stage. Math1 showed the strongest expression. Our results suggest that the Notch1 signaling pathway through Jagged2 could be importantly related to Math1, directing the process of odontogenesis toward cell differentiation.
Notch信号通路是一种进化保守的机制,在多种组织的细胞间通讯和细胞命运中起着重要作用。哺乳动物Notch受体家族包括4个成员:Notch1/2/3/4。Notch配体家族由5个成员组成:Delta1/3/4和Jagged1/2。Math1编码小鼠基本螺旋-环-螺旋(bHLH)转录因子,作为细胞分化的正调节因子。最近,Notch和Math1通路之间的联系在各种组织中得到证实。采用原位杂交技术分析了胚胎期(E) 13和E15以及产后期(PN) 1、PN3、PN5、PN10和PN14小鼠磨牙胚中Notch1、Jagged2和Math1的表达。Notch1在牙胚期的牙芽中表达阳性,但在牙间质接触的基底细胞中不表达。Jagged2和Math1在分化成釉细胞和成牙细胞中强表达,Math1的强表达甚至维持到PN14期。Math1表达最强。我们的研究结果表明,通过Jagged2的Notch1信号通路可能与Math1有重要关系,指导牙形成过程走向细胞分化。
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引用次数: 9
Agmatine pretreatment protects retinal ganglion cells (RGC-5 cell line) from oxidative stress in vitro. 胍丁胺预处理对视网膜神经节细胞(RGC-5细胞系)氧化应激的保护作用。
Y. Iizuka, Samin Hong, C. Kim, S. Kim, G. Seong
Agmatine, 2-(4-aminobutyl)guanidine, has been reported to have neuroprotective effects against various neuronal damages. In this study it was investigated whether agmatine pretreatment rescues the retinal ganglion cells from oxidative injury in vitro. After differentiation of transformed rat retinal ganglion cells (RGC-5 cell line) with staurosporine, agmatine (0.0 to 100.0 microM) pretreatment was performed for 2 hours. Subsequently, they were exposed to hydrogen peroxide (0.0 to 2.5 mM) as an oxidative stress. Cell viability was monitored for up to 48 hours with the lactate dehydrogenase (LDH) assay and apoptosis was examined by the terminal deoxynucleotide transferase-mediated terminal uridine deoxynucleotidyl transferase nick end-labeling (TUNEL) method. As a result, differentiated RGC-5 cells were found to have decreased viability after addition of hydrogen peroxide in a dose-dependent manner. This hydrogen peroxide induced cytotoxicity caused apoptosis characterized by DNA fragmentation. Agmatine pretreatment not only increased cell viability but also attenuated DNA fragmentation. In conclusion, agmatine pretreatment demonstrated neuroprotective effects against oxidative stress induced by hydrogen peroxide in differentiated RGC-5 cells in vitro. This suggests a novel therapeutic strategy rescuing retinal ganglion cells from death caused by oxidative injury.
胍丁氨酸,2-(4-氨基丁基)胍,已被报道对各种神经损伤具有神经保护作用。本研究探讨了胍丁胺预处理是否能在体外修复视网膜神经节细胞的氧化损伤。转化后的大鼠视网膜神经节细胞(RGC-5细胞系)经staurosporine分化后,agmatine(0.0 ~ 100.0微米)预处理2小时。随后,将它们暴露于过氧化氢(0.0至2.5 mM)中作为氧化应激。采用乳酸脱氢酶(LDH)法监测细胞活力长达48小时,采用末端脱氧核苷酸转移酶介导的末端尿苷脱氧核苷酸转移酶缺口末端标记(TUNEL)法检测细胞凋亡。结果发现,在加入过氧化氢后,分化的RGC-5细胞的活力呈剂量依赖性下降。过氧化氢诱导细胞毒性导致以DNA断裂为特征的细胞凋亡。Agmatine预处理不仅能提高细胞活力,还能减弱DNA断裂。综上所述,胍丁胺预处理对体外分化的RGC-5细胞具有抗过氧化氢氧化应激的神经保护作用。这提示了一种新的治疗策略,可以挽救氧化损伤引起的视网膜神经节细胞死亡。
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引用次数: 22
DNA mismatch repair gene methylation in gastric cancer in individuals from northern Brazil. 巴西北部个体胃癌DNA错配修复基因甲基化。
E. Moura Lima, M. Ferreira Leal, M. C. Cardoso Smith, R. R. Rodriguez Burbano, P. Pimentel de Assumpção, M. J. Bello, J. Rey, Francinaldo Ferreira de Lima, C. Casartelli
Gastric cancer is one of the most common malignancies. DNA methylation is implicated in DNA mismatch repair genes deficiency. In the present study, we evaluated the methylation status of MLH1, MSH2, MSH6 and PMS2 in 20 diffuse- and 26 intestinal-type gastric cancer samples and 20 normal gastric mucosal of gastric cancer patients from Northern Brazil. We found that none of the nonneoplastic samples showed methylation of any gene promoter and 50% of gastric cancer samples showed at least one methylated gene promoter. Methylation frequencies of MLH1, MSH2, MSH6 and PMS2 promoter were 21.74%, 17.39%, 0% and 28.26% respectively in gastric cancer samples. MLH1 and PMS2 methylation were associated with neoplastic samples compared to nonneoplastic ones. PMS2 methylation was associated with diffuse- and intestinal-type cancer compared with normal controls. Intestinal-type cancer showed significant association with MLH1 methylation. Diffuse-type cancer was significantly associated with MSH2 methylation. Our findings show differential gene methylation in tumoral tissue, which allows us to conclude that methylation is associated with gastric carcinogenesis. Methylation of mismatch repair genes was associated with gastric carcinogenesis and may be a helpful tool for diagnosis, prognosis and therapies. However, MSH6 does not seem to be regulated by methylation in our samples.
胃癌是最常见的恶性肿瘤之一。DNA甲基化与DNA错配修复基因缺陷有关。在本研究中,我们评估了来自巴西北部胃癌患者的20例弥漫性胃癌和26例肠型胃癌样本以及20例正常胃粘膜中MLH1、MSH2、MSH6和PMS2的甲基化状态。我们发现,没有任何非肿瘤样本显示任何基因启动子甲基化,50%的胃癌样本显示至少一个甲基化的基因启动子。胃癌样本中MLH1、MSH2、MSH6和PMS2启动子甲基化频率分别为21.74%、17.39%、0%和28.26%。与非肿瘤样品相比,MLH1和PMS2甲基化与肿瘤样品相关。与正常对照相比,PMS2甲基化与弥漫性癌和肠型癌相关。肠型癌与MLH1甲基化显著相关。弥漫性癌与MSH2甲基化显著相关。我们的研究结果显示肿瘤组织中存在差异基因甲基化,这使我们能够得出甲基化与胃癌发生有关的结论。错配修复基因的甲基化与胃癌的发生有关,可能是诊断、预后和治疗的有用工具。然而,在我们的样本中,MSH6似乎不受甲基化的调节。
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引用次数: 27
Agmatine inhibits hypoxia-induced TNF-alpha release from cultured retinal ganglion cells. 胍丁胺抑制缺氧诱导的视网膜神经节细胞tnf - α释放。
Samin Hong, Kyoungsoo Park, C. Kim, G. Seong
The effect of hypoxia on the release of tumor necrosis factor-alpha (TNF-alpha) in transformed rat retinal ganglion cells (RGCs) and the effect of agmatine on the hypoxia-induced production of TNF-alpha in RGCs were evaluated. RGCs were cultured under hypoxic conditions with 5% oxygen, with or without 100 microM agmatine. The expression levels of TNF-alpha and its receptor-1 (TNF-R1) were investigated by Western blot analysis. After 6 hours of hypoxia, we noted an increase in TNF-alpha production in RGCs. Agmatine significantly reduced TNF-alpha level after 12 hours of hypoxic treatment. The expression of TNF-R1 was not affected by the hypoxia or agmatine treatment. Our results show that agmatine inhibits the TNF-alpha production of RGCs in hypoxic condition. These results demonstrate a possible neuroprotective mechanism for agmatine against hypoxic damage in RGCs.
研究了缺氧对转化大鼠视网膜神经节细胞(RGCs)肿瘤坏死因子- α (tnf - α)释放的影响以及胍丁氨酸对缺氧诱导的RGCs肿瘤坏死因子- α生成的影响。RGCs在5%氧气的低氧条件下培养,加或不加100 μ m胍。Western blot检测tnf - α及其受体1 (TNF-R1)的表达水平。缺氧6小时后,我们注意到RGCs中tnf - α的产生增加。Agmatine在缺氧治疗12小时后显著降低tnf - α水平。TNF-R1的表达不受缺氧和agmatine处理的影响。我们的研究结果表明,agmatine在缺氧条件下抑制RGCs的tnf - α产生。这些结果表明,agmatine可能具有抗RGCs缺氧损伤的神经保护机制。
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引用次数: 24
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Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al
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