Soriano E, Del Rio J A
Unidad de Biologia Celular, Facultad de Biologia, Universidad de Barcelona, Spain.
J Histochem Cytochem. 1991 Mar;39(3):255-63. doi: 10.1177/39.3.1671576.
5'-Bromodeoxyuridine (BrdU) is a thymidine analogue which can be detected by monoclonal antibodies (MAb). We have developed a method for the simultaneous visualization of BrdU and a wide range of neural antigens in paraformaldehyde-fixed brain sections. Pregnant mice were injected intraperitoneally with a single pulse of BrdU. Young adult offspring were processed for immunocytochemistry following a double immunoperoxidase sequence. BrdU was detected using diaminobenzidine (DAB) intensified with nickel ammonium sulfate and neural antigen-containing elements were visualized with DAB alone. BrdU-positive nuclei and tissue antigen-immunoreactive cells were easily differentiated. Furthermore, double-labeled cells characterized by the presence of a black immunoreactive nucleus surrounded by a brown immunopositive cytoplasm were unambiguously recognized. Satisfactory results were obtained using either MAb or polyclonal antibodies against a variety of cell antigens, including neuropeptides, CA++ binding proteins, and cytoskeletal components of the glial cells. The method reported here permits analysis of the neurogenesis and proliferation of subsets of neurons and glial cells, identified by immunocytochemical markers.
5'-溴脱氧尿苷（BrdU）是一种胸腺嘧啶类似物，可通过单克隆抗体（MAb）进行检测。我们开发了一种在多聚甲醛固定的脑切片中同时可视化BrdU和多种神经抗原的方法。给怀孕小鼠腹腔内注射单次脉冲的BrdU。成年幼崽按照双重免疫过氧化物酶序列进行免疫细胞化学处理。使用硫酸镍铵强化的二氨基联苯胺（DAB）检测BrdU，仅用DAB可视化含神经抗原的成分。BrdU阳性细胞核和组织抗原免疫反应性细胞易于区分。此外，以黑色免疫反应性细胞核被棕色免疫阳性细胞质包围为特征的双标记细胞也能被明确识别。使用针对多种细胞抗原（包括神经肽、Ca++结合蛋白和神经胶质细胞的细胞骨架成分）的单克隆抗体或多克隆抗体均获得了满意的结果。本文报道的方法允许对通过免疫细胞化学标记物鉴定的神经元和神经胶质细胞亚群的神经发生和增殖进行分析。
