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Cytochemical and ultrastructural study of anoikis and secondary necrosis in enterocytes detached in vivo

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Abstract

Detachment-induced apoptosis of enterocytes (anoikis) has not been investigated in vivo. Here we describe anoikis of fish enterocytes following detachment in a septicemia by Photobacterium damselae subsp. piscicida, or following injection of its exotoxin. The in vivo study was complemented with an ex vivo time-lapse analysis using conditions duplicating the in vivo situation. Linings of enterocytes detached from intestine mucosa dissociate into isolated enterocytes which undergo caspase 3-mediated anoikis with cell rounding, loss of polarization, condensation of chromatin and fragmentation of the nuclear envelope, early swelling of mitochondria with rupture of the outer membrane, and brush border disappearance. One mechanism for brush border loss was shedding of apoptotic bodies incorporating the apical part of the enterocyte. Brush border disappearance was also associated with disassembly of the F-actin microvillar core and involved re-absorption into the cell, or expansion and vesiculation followed by shedding of microvillar fragments. The enterocyte anoikis terminates by secondary necrosis and lysis due to lack of elimination by phagocytosis of apoptosing enterocytes. The conditions prevailing in vivo in the gut lumen accelerate enterocyte secondary necrosis. Our results underscore the importance of analyzing anoikis under conditions similar to those occurring in vivo.

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Acknowledgments

The authors are grateful to Goreti Pinto, Alexandra Rema, Célia Lopes and Fátima Faria for assistance with histology. This work was supported by FCT (Fundação para a Ciência e Tecnologia) project POCI/MAR/56111/2004 funded by POCI 2010 and co-funded by FEDER, and grants SFRH/BPD/26816/2006 and SFRH/BPD/20881/2004.

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Authors and Affiliations

  1. IBMC—Instituto de Biologia Molecular e Celular, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal

    Ana do Vale, Carolina Costa-Ramos, Daniela S. P. Silva, Paula M. Macedo, Rui Fernandes, Paula Sampaio, Nuno M. S. dos Santos & Manuel T. Silva

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  1. Ana do Vale
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  2. Carolina Costa-Ramos
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Correspondence to Manuel T. Silva.

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do Vale, A., Costa-Ramos, C., Silva, D.S.P. et al. Cytochemical and ultrastructural study of anoikis and secondary necrosis in enterocytes detached in vivo . Apoptosis 12, 1069–1083 (2007). https://doi.org/10.1007/s10495-006-0040-x

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