Ferrer I, Planas A M, Pozas E
Unitat de Neuropatologia, Servei d'Anatomia Patològica, Hospital Princeps d'Espanya, Universitat de Barcelona, Spain.
Neuroscience. 1997 Sep;80(2):449-58. doi: 10.1016/s0306-4522(97)00126-7.
Ionizing radiation produces apoptosis in the developing rat brain. Strong c-Jun immunoreactivity, as revealed with the antibody c-Jun/AP-1 (N) which is raised against the amino acids 91-105 mapping with the amino terminal domain of mouse c-Jun p39, is simultaneously observed in the nucleus and cytoplasm of apoptotic cells. Western blotting of total brain homogenates, using the same antibody, shows a p39 band in control rats which is accompanied by a strong, phosphorylated p62 double-band in irradiated animals. In addition, increased c-Jun N-terminal kinase 1 expression, as found on western blots, is found in irradiated rats when compared with controls. Intraperitoneal injection of kainic acid at convulsant doses to the adult rat produces cell death with morphological features of necrosis, together with the appearance of cells with fine granular chromatin degeneration and small numbers of apoptotic-like cells, in the entorhinal and piriform cortices, basal amygdala, certain thalamic nuclei, and CA1 region of the hippocampus. c-Jun expression in kainic acid-treated rats, as revealed with the c-Jun/AP-1 (N) antibody, is found in the nuclei of a minority of cells in the same areas. The vast majority of c-Jun-immunoreactive cells have normal nuclear morphology, whereas necrotic cells are negative and only a few cells with fine granular chromatin condensation and apoptotic cells following kainic acid injection are stained with c-Jun antibodies. Western blotting, using the same antibody, shows a p39 band in control rats, which is accompanied by a band at about p26 from 6 h onwards following kainic acid injection. Decreased c-Jun N-terminal kinase 1 expression, as revealed on western blots, is observed in kainic acid-treated rats. These results show that the antibody c-Jun/AP-1 (N) recognizes three different forms of c-Jun-related immunoreactivity in normal and pathological states, which are associated with the different outcome of cells. These results stress the necessity of examining in detail the composition of c-Jun-immunoreactive bands and the metabolic state of c-Jun(s) in different paradigms of cell death and survival.
电离辐射可导致发育中的大鼠大脑发生细胞凋亡。用针对小鼠c-Jun p39氨基末端结构域中91-105位氨基酸产生的抗体c-Jun/AP-1 (N)检测发现，凋亡细胞的细胞核和细胞质中同时出现强c-Jun免疫反应性。使用相同抗体对全脑匀浆进行蛋白质印迹分析显示，对照大鼠中有一条p39条带，而受辐照动物中除p39条带外，还伴有一条强的、磷酸化的p62双条带。此外，蛋白质印迹分析发现，与对照相比，受辐照大鼠中c-Jun氨基末端激酶1的表达增加。给成年大鼠腹腔注射惊厥剂量的海藻酸可导致细胞死亡，伴有坏死的形态学特征，在内嗅皮质、梨状皮质、基底杏仁核、某些丘脑核以及海马体CA1区出现具有细颗粒状染色质变性的细胞和少量凋亡样细胞。用c-Jun/AP-1 (N)抗体检测发现，海藻酸处理的大鼠中，同一区域少数细胞的细胞核中有c-Jun表达。绝大多数c-Jun免疫反应性细胞具有正常的核形态，而坏死细胞为阴性，海藻酸注射后只有少数具有细颗粒状染色质凝聚的细胞和凋亡细胞被c-Jun抗体染色。使用相同抗体进行蛋白质印迹分析显示，对照大鼠中有一条p39条带，海藻酸注射后6小时起出现一条约p26的条带。蛋白质印迹分析显示，海藻酸处理的大鼠中c-Jun氨基末端激酶1的表达降低。这些结果表明，抗体c-Jun/AP-1 (N)可识别正常和病理状态下三种不同形式的c-Jun相关免疫反应性，它们与细胞的不同结局相关。这些结果强调了在不同的细胞死亡和存活模式中详细检查c-Jun免疫反应性条带的组成和c-Jun的代谢状态的必要性。
