Biology Department, Wesleyan University, Middletown, CT 06459, USA

Correspondence to: J Wolfe, Biology Department, Wesleyan University, Middletown, CT 06459, USA. Tel: 860 685-3239; Fax: 860 685-3279; E-mail: jwolfe@wesleyan.edu

Edited by L Schwartz

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. Cell Death and Differentiation (2001) 8, 289-297.

acidification; apoptosis; autophagosome; ciliate; lysosome; nucleus

PCD, programmed cell death; AO, acridine orange; LB, lysosomal bodies; DAPI, 4,6-diamidino-2-phenylindole; TUNEL, Tdt-mediated dUTP nick end-labeling