Lu E, Wolfe J
Biology Department, Wesleyan University, Middletown, CT 06459, USA.
Cell Death Differ. 2001 Mar;8(3):289-97. doi: 10.1038/sj.cdd.4400807.
A key characteristic of apoptosis is its regulated nuclear degradation. Apoptosis-like nuclear degradation also occurs in the ciliated unicellular organism, Tetrahymena thermophila. Chromatin of the macronucleus undergoes massive condensation, a process that can be blocked by caspase inhibitors. The nucleus becomes TUNEL-positive, and its DNA is cleaved into nucleosome-sized fragments. In a matter of hours the macronucleus is completely degraded, and disappears. The condensed nucleus sequesters acridine orange, which means that it might become an acidic compartment. We therefore asked whether lysosomal bodies fuse with the condensed macronucleus to form an autophagosome. We monitored acid phosphatase (AP) activity, which is associated with lysosomal bodies but is not found in normal nuclei. We find that after the macronucleus condenses AP activity is localized in cap-like structures at its cortex. Later, after the degrading macronucleus loses much of its DNA, acid phosphatase deposits appear deeper within the nucleus. We conclude that although macronuclear elimination is initiated by an apoptosis-like mechanism, its final degradation may be achieved through autophagosomy.
细胞凋亡的一个关键特征是其受调控的核降解。类似细胞凋亡的核降解也发生在纤毛单细胞生物嗜热栖热放线菌中。大核的染色质会发生大量浓缩,这一过程可被半胱天冬酶抑制剂阻断。细胞核变为TUNEL阳性,其DNA被切割成核小体大小的片段。在数小时内,大核会完全降解并消失。浓缩的细胞核会摄取吖啶橙,这意味着它可能变成一个酸性区室。因此,我们询问溶酶体是否会与浓缩的大核融合形成自噬体。我们监测了酸性磷酸酶(AP)的活性,该活性与溶酶体相关,但在正常细胞核中不存在。我们发现,大核浓缩后,AP活性定位于其皮质的帽状结构中。之后,在降解的大核失去大量DNA后,酸性磷酸酶沉积物出现在细胞核更深的位置。我们得出结论,虽然大核的消除是由类似细胞凋亡的机制启动的,但其最终降解可能是通过自噬体实现的。