Penfold M E, Armati P J, Mikloska Z, Cunningham A L
Virology Department, University of Sydney, ICPMR, Westmead Hospital, Sydney, Australia.
In Vitro Cell Dev Biol Anim. 1996 Jul-Aug;32(7):420-6. doi: 10.1007/BF02723004.
The interaction of autologous human fetal neurons with epidermal cells was studied by culturing fetal dorsal root ganglia (DRG) in the center of a dual chamber system with epidermal explants in the outer chamber. The two chambers were separated by two concentric stainless steel annular rings adherent to the substratum by silicon grease and agarose. Axons from the DRG penetrated the agarose barrier, growing into the exterior chamber by 10 d in vitro (DIV) and extended past sparse peripheral fibroblasts to interact specifically with epidermal cells by 12 to 16 DIV. Scanning electron microscopy (SEM) showed single or multiple neuronal fascicles terminating on epidermal cells with spatular, veillike or bulbous axon termini. Transmission electron microscopy (TEM) showed fine axonal termini between epidermal cells, separated by an intercellular gap. The specificity of axonal targeting for epidermal cells rather than fibroblasts was also demonstrated by infecting the DRG with Herpes simplex virus type-1 (HSV-1). Specific anterograde transport of HSV-1 along axons to keratin-expressing epidermal cells was demonstrated by immunofluorescence and immunoperoxidase staining using monoclonal antibodies to viral glycoprotein D. This model allows the study of the mechanism of the specific interactions between neurons and epidermal cells analogous to those in fetal development and after cutaneous nerve regeneration.
通过将胎儿背根神经节（DRG）培养在双室系统的中心，在外室培养表皮外植体，研究了自体人胎儿神经元与表皮细胞的相互作用。两个腔室由两个同心不锈钢环形环隔开，环形环通过硅脂和琼脂糖附着在基质上。来自DRG的轴突穿透琼脂糖屏障，在体外培养10天（DIV）时生长到外室，并在12至16 DIV时延伸越过稀疏的外周成纤维细胞，与表皮细胞特异性相互作用。扫描电子显微镜（SEM）显示单个或多个神经束终止于具有铲状、面纱状或球状轴突末端的表皮细胞上。透射电子显微镜（TEM）显示表皮细胞之间有精细的轴突末端，由细胞间隙隔开。用1型单纯疱疹病毒（HSV-1）感染DRG也证明了轴突靶向表皮细胞而非成纤维细胞的特异性。使用针对病毒糖蛋白D的单克隆抗体，通过免疫荧光和免疫过氧化物酶染色证明了HSV-1沿轴突向表达角蛋白的表皮细胞的特异性顺行运输。该模型允许研究神经元与表皮细胞之间特异性相互作用的机制，类似于胎儿发育和皮肤神经再生后的机制。
