Feigenspan A, Bormann J, Wässle H
Max-Planck Institut für Hirnforschung, Neuroanatomische Abteilung, Frankfurt, Germany.
Vis Neurosci. 1993 Mar-Apr;10(2):203-17. doi: 10.1017/s0952523800003618.
Vertical slices of 6-day postnatal (P6) rat retina were cut at a thickness of 100 microns and cultured using the roller-tube technique. After 14-21 days in vitro there was significant distortion of normal retinal architecture, but localized areas of the slices showed the typical pattern of layering of mature retina. The following immunocytochemical markers were used to characterize the different retinal cell types: antibodies against protein kinase C (PKC), calcium binding protein (CabP 28kD), neurofilaments (NF), glia-specific antibodies (GFAP, vimentin), and transmitter-specific antibodies (GABA, TH). The expression of these markers was compared in P6 retina, adult retina, and slice culture. To further characterize the cultured cells, patch-clamp recordings were performed in combination with intracellular injection of Lucifer Yellow (LY). Transmitter- and voltage-gated membrane currents were recorded from morphologically identified neurons. The experiments show that a mammalian slice culture can be used to study differentiation and function of retinal cell types.
将出生后6天（P6）的大鼠视网膜切成100微米厚的垂直切片，并采用滚管技术进行培养。体外培养14 - 21天后，正常视网膜结构出现明显扭曲，但切片的局部区域呈现出成熟视网膜典型的分层模式。使用以下免疫细胞化学标记物来表征不同的视网膜细胞类型：抗蛋白激酶C（PKC）抗体、钙结合蛋白（CabP 28kD）、神经丝（NF）、神经胶质特异性抗体（GFAP、波形蛋白）以及递质特异性抗体（GABA、TH）。比较了这些标记物在P6视网膜、成年视网膜和切片培养物中的表达情况。为了进一步表征培养的细胞，结合细胞内注射荧光黄（LY）进行了膜片钳记录。从形态学上鉴定的神经元记录了递质门控和电压门控膜电流。实验表明，哺乳动物切片培养可用于研究视网膜细胞类型的分化和功能。
