Mommaas M, Tada J, Ponec M
Department of Dermatology, University Hospital Leiden, The Netherlands.
J Dermatol Sci. 1991 Mar;2(2):97-105. doi: 10.1016/0923-1811(91)90018-s.
Immunoelectronmicroscopy was employed to investigate the ultrastructural localization of the low density lipoprotein (LDL) receptor and the protein moiety of LDL (apo B) on normal human epidermis in situ and reconstructed human epidermis in vitro. For this purpose ultrathin sections of Lowicryl K4M embedded material were incubated with monoclonal antibodies against the LDL receptor and apo B followed by a second antibody conjugated to 15 nm colloidal gold. Examination of the sections revealed a similar distribution of the LDL receptor and apo B in normal human epidermis and in reconstructed epidermis. In both systems the amount of LDL receptors decreases during keratinocyte differentiation. In contrast, apo B molecules are more abundantly expressed in the upper layers of the stratum spinosum and the stratum granulosum. The great similarities in the distribution of the LDL receptor and apo B between the in vitro reconstructed epidermis and its in vivo counterpart provide additional proof that reconstructed epidermis is an excellent tool to investigate proliferation and differentiation processes of keratinocytes in vitro.
采用免疫电子显微镜技术,研究低密度脂蛋白(LDL)受体和LDL蛋白部分(载脂蛋白B)在正常人原位表皮和体外重建人表皮中的超微结构定位。为此,将Lowicryl K4M包埋材料的超薄切片与抗LDL受体和载脂蛋白B的单克隆抗体孵育,然后与结合15 nm胶体金的二抗孵育。切片检查显示,LDL受体和载脂蛋白B在正常人表皮和重建表皮中的分布相似。在这两个系统中,角质形成细胞分化过程中LDL受体的数量减少。相反,载脂蛋白B分子在棘层和颗粒层上层表达更为丰富。体外重建表皮与其体内对应物之间LDL受体和载脂蛋白B分布的高度相似性,进一步证明重建表皮是体外研究角质形成细胞增殖和分化过程的优良工具。