Castel M, Morris J, Ben-Barak Y, Timberg R, Sivan N, Gainer H
J Histochem Cytochem. 1985 Oct;33(10):1015-25. doi: 10.1177/33.10.3900193.
Using three different monoclonal antibodies against rat neurophysins (5), with protein A-gold as immunocytochemical marker (27), the murid hypothalamoneurohy-pophysial system was studied at the ultrastructural level. Postembedding staining was done on epoxy-embedded sections of supraoptic nuclei and posterior pituitaries. Specific immunolabeling of vasopressinergic and oxytocinergic neurosecretory granules was observed in tissues fixed with glutaraldehyde or glutaraldehyde mixtures (containing paraformaldehyde and picric acid), with or without osmium tetroxide postfixation and with or without sodium metaperiodate oxidation. Some autophagic vacuoles containing lysed neurosecretory granules were also neurophysin immunoreactive. Nonspecific background staining was extremely low. An attempt was made to appraise labeling intensities semiquantitatively by counting gold particles in relation to number of secretory granules per axonal varicosity. Immunoreactivity was measurably influenced by the mode of fixation, sodium metaperiodate oxidation, and titer and affinity of the antibody. The protein A-gold technique using monoclonal antibodies against neurophysins provides a superior means of ultrastructural analysis of the hypothalamoneurohypophysial system, both visually and morphometrically.
使用三种针对大鼠神经垂体素的不同单克隆抗体(5)，以蛋白A-金作为免疫细胞化学标记物(27)，在超微结构水平上研究了啮齿动物下丘脑神经垂体系统。对视上核和垂体后叶的环氧树脂包埋切片进行包埋后染色。在用戊二醛或戊二醛混合物(含多聚甲醛和苦味酸)固定的组织中，无论是否进行四氧化锇后固定以及是否进行高碘酸钠氧化，均观察到血管加压素能和催产素能神经分泌颗粒的特异性免疫标记。一些含有溶解的神经分泌颗粒的自噬泡也具有神经垂体素免疫反应性。非特异性背景染色极低。试图通过计算相对于每个轴突膨体分泌颗粒数量的金颗粒来半定量评估标记强度。免疫反应性受到固定方式、高碘酸钠氧化以及抗体效价和亲和力的显著影响。使用针对神经垂体素的单克隆抗体的蛋白A-金技术，在视觉和形态计量学方面都为下丘脑神经垂体系统的超微结构分析提供了一种优越的方法。
