Buniatian G, Gebhardt R, Schrenk D, Hamprecht B
Physiologisch-chemisches Institut, Universität Tübingen, Germany.
Eur J Cell Biol. 1996 May;70(1):23-32.
The presence and the colocalization of the three intermediate filament proteins, glial fibrillary acidic protein (GFAP) and the marker of mesenchymal liver cells, vimentin, were studied by an immunofluorescence double-labeling technique in cultures of isolated rat perisinusoidal stellate cells (PSC) and hepatocytes, in cocultures of isolated PSC and hepatocytes as well as in cryostat sections of rat liver. GFAP and vimentin immunoreactivities were localized in cultured PSC which were identified by the presence of the cellular marker desmin, another intermediate filament protein, or the stellate morphology to be seen after staining for one of three intermediate filament proteins. Both GFAP and vimentin were strongly expressed in the perinuclear region and the cell processes of cultured PSC. Staining for GFAP highly coincided with that for vimentin or desmin in cultured PSC and with that for vimentin in the liver sections. Desmin-positive cells were always also GFAP-positive. However, of the GFAP-positive cells only an estimated 50% were found desmin-positive. The coexpression of desmin and GFAP in the same cells appear to be unique, since apparently it has not been previously reported for any other cell type. Almost all of the vimentin-positive cells in hepatocyte culture were also expressing GFAP. Since desmin was not found in all of the cultured cells with PSC morphology, GFAP is suggested as a more reliable marker for PSC than desmin.
采用免疫荧光双标记技术,在分离的大鼠肝血窦周星状细胞(PSC)和肝细胞培养物中、分离的PSC与肝细胞的共培养物以及大鼠肝脏冰冻切片中,研究了三种中间丝蛋白——胶质纤维酸性蛋白(GFAP)和间充质肝细胞标志物波形蛋白的存在及共定位情况。GFAP和波形蛋白免疫反应性定位于培养的PSC中,这些PSC通过细胞标志物结蛋白(另一种中间丝蛋白)的存在或在对三种中间丝蛋白之一进行染色后可见的星状形态来鉴定。GFAP和波形蛋白在培养的PSC的核周区域和细胞突起中均强烈表达。在培养的PSC中,GFAP染色与波形蛋白或结蛋白的染色高度一致,在肝脏切片中与波形蛋白的染色高度一致。结蛋白阳性细胞总是也为GFAP阳性。然而,在GFAP阳性细胞中,仅估计50%的细胞为结蛋白阳性。结蛋白和GFAP在同一细胞中的共表达似乎是独特的,因为此前显然尚未在任何其他细胞类型中报道过。在肝细胞培养物中,几乎所有波形蛋白阳性细胞也表达GFAP。由于在所有具有PSC形态的培养细胞中未发现结蛋白,因此GFAP被认为是比结蛋白更可靠的PSC标志物。