Vedrenne Nicolas, Sarrazy Vincent, Richard Laurence, Bordeau Nelly, Battu Serge, Billet Fabrice, Desmoulière Alexis
EA 6309 "Myeline maintenance and peripheral neuropathies", University of Limoges, 87000, Limoges, France.
Mediterranean Centre for Molecular Medicine, University of Nice Sophia Antipolis, INSERM U1065, 06000, Nice, France.
Neurochem Res. 2017 Aug;42(8):2427-2434. doi: 10.1007/s11064-017-2268-y. Epub 2017 Apr 22.
A wide heterogeneity of lesions can affect the central nervous system (CNS). In all situations where neurons are damaged, including multiple sclerosis (MS), a common reactive astrocytosis is present. Sedimentation field-flow fractionation (SdFFF) was used to sort astrocyte subpopulations. After SdFFF elution, cells, prepared from rat newborn cortex, were cultured and analyzed by immunocytofluorescence for glial fibrillary acidic protein (GFAP) and α-smooth muscle (SM) actin (a specific marker for myofibroblasts) expression. Cell contractile capacity was studied. Samples from patients with MS were also analyzed. Three main fractions (F1, F2, and F3) were isolated and compared with the total eluted population (TP). TP, F1, F2, and F3, contained respectively 74, 96, 12, and 98% of GFAP expressing astrocytes. In F3, astrocytes only expressed GFAP while in F1, astrocytes expressed both GFAP and α-SM actin. In F2 and TP, α-SM actin expression was barely detected. F3-derived cells showed higher contractile capacities compared with F1-derived cells. In one specific case of MS known as Baló's concentric MS, astrocytes expressing both GFAP and α-SM actin were detected. Using SdFFF, a population of astrocytes presenting myofibroblast properties was isolated. This subpopulation of astrocytes was also observed in a MS sample suggesting that it could be involved in lesion formation and remodeling during CNS pathologies.
多种病变的广泛异质性可影响中枢神经系统(CNS)。在所有神经元受损的情况下,包括多发性硬化症(MS),都会出现常见的反应性星形胶质细胞增生。沉降场流分级分离法(SdFFF)用于分选星形胶质细胞亚群。在SdFFF洗脱后,培养从新生大鼠皮层制备的细胞,并通过免疫细胞荧光分析胶质纤维酸性蛋白(GFAP)和α-平滑肌(SM)肌动蛋白(肌成纤维细胞的特异性标志物)的表达。研究了细胞收缩能力。还分析了MS患者的样本。分离出三个主要组分(F1、F2和F3),并与总洗脱群体(TP)进行比较。TP、F1、F2和F3分别含有表达GFAP的星形胶质细胞的74%、96%、12%和98%。在F3中,星形胶质细胞仅表达GFAP,而在F1中,星形胶质细胞同时表达GFAP和α-SM肌动蛋白。在F2和TP中,几乎检测不到α-SM肌动蛋白的表达。与F1来源的细胞相比,F3来源的细胞表现出更高的收缩能力。在一种称为巴洛同心性MS的特定MS病例中,检测到同时表达GFAP和α-SM肌动蛋白的星形胶质细胞。使用SdFFF,分离出一群具有肌成纤维细胞特性的星形胶质细胞。在一个MS样本中也观察到了这种星形胶质细胞亚群,这表明它可能参与了CNS病变过程中的病变形成和重塑。