Author:
Date:
2015-06-20
[Abstract] Lipid droplets (LDs) are ubiquitous intracellular structures whose formation, growth, and maintenance are highly regulated (Wang et al., 2013; Ranall et al., 2011; Goodman, 2009). Lipid metabolism and droplet dynamics are of considerable interest to agriculture, biofuel production, viral pathology, nutrition, and cancer biology (Walther and Farese, 2009; Liu et al., 2010). Accumulation of fatty acids and neutral lipids in nonadipose tissues is cytotoxic (Kourtidis et al., 2009). BODIPY 493/503 (4,4-Difluoro-1,3,5,7,8-Pentamethyl-4-Bora-3a,4a-Diaza-s-Indacene) is the standard dye to study LDs within adipocytes. BODIPY 493/503 contains a nonpolar structure that, upon binding to neutral lipid, emits a green fluorescence signal with a narrow wavelength ...
[摘要] 脂质滴(LD)是普遍存在的细胞内结构,其形成,生长和维持是高度调节的(Wang等人,2013; Ranall等人,2011; Goodman, 2009)。脂质代谢和液滴动力学对农业,生物燃料生产,病毒病理学,营养和癌症生物学是相当感兴趣的(Walther和Farese,2009; Liu等人,2010)。在非脂肪组织中脂肪酸和中性脂质的积累是细胞毒性的(Kourtidis等人,2009)。 BODIPY 493/503(4,4-二氟-1,3,5,7,8-五甲基-4-Bora-3a,4a-Diaza-s-Indacene)是研究脂肪细胞内LD的标准染料。 BODIPY 493/503含有非极性结构,在结合中性脂质时,发射具有窄波长范围的绿色荧光信号,使其成为用于多标记实验的理想荧光团。染料分子的疏水性质促进快速进入LD的非极性环境(Listenberge和Brown,2007)。 Gocze和Freeman显示,当使用BODIPY493/503与Nile Red相比时,脂质荧光变异性显着降低,表明其可能对LD更具特异性(Gocze和Freeman,1994)。在这里,我们描述了BODIPY 493/503测定用于检测培养的细胞中的神经脂肪储存(图1)(Wang等人,2013)。
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Author:
Date:
2014-01-20
[Abstract] The ubiquitin-proteasome system (UPS) mediates the majority of the proteolysis seen in the cytoplasm and nucleus of mammalian cells. As such it plays an important role in the regulation of a variety of physiological and pathophysiological processes including tumorigenesis, inflammation and cell death (Ciechanover, 2005; Kisselev and Goldberg, 2001). A number of recent studies have shown that proteasome activity is decreased in a variety of neurological disorders including Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis and stroke as well as during normal aging (Chung et al., 2001; Ciechanover and Brundin, 2003; Betarbet et al., 2005). This decrease in proteasome activity is thought to play a critical role in the accumulation of abnormal and ...
[摘要] 泛素 - 蛋白酶体系统(UPS)介导哺乳动物细胞的细胞质和细胞核中大部分蛋白水解。因此,它在调节各种生理和病理生理过程中起重要作用,包括肿瘤发生,炎症和细胞死亡(Ciechanover,2005; Kisselev和Goldberg,2001)。许多最近的研究表明,在多种神经障碍(包括帕金森病,阿尔茨海默病,肌萎缩性侧索硬化和中风)以及正常衰老期间,蛋白酶体活性降低(Chung等,2001; Ciechanover和Brundin,2003; Betarbet等,2005)。蛋白酶体活性的降低被认为在异常和氧化蛋白质的积累中发挥关键作用。 UPS的蛋白质清除涉及两个连续的反应。第一个是用泛素(Ub)标记蛋白质赖氨酸残基,第二个是蛋白酶体随后降解标记的蛋白质。我们本文描述了这两个反应中第二个的测定(Valera等,2013)。该测定法使用荧光底物作为蛋白酶体的三种活性:糜蛋白酶样活性,胰蛋白酶样活性和半胱天冬酶样活性。通过蛋白酶体从底物中切割荧光团可导致可用荧光板读数器检测的荧光。
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