1887

Abstract

This paper describes the serendipitous discovery and first characterization of a new resistant cell type from , for which the name aspidocyte (from : Greek for shield) is proposed. These cells are induced from amoebae by a range of toxins including heavy metals and antibiotics, and were first detected by their striking resistance to detergent lysis. Aspidocytes are separate, rounded or irregular-shaped cells, which are immotile but remain fully viable; once the toxic stress is removed, they revert to amoeboid cells within an hour. Induction takes a few hours and is completely blocked by the protein synthesis inhibitor cycloheximide. Aspidocytes lack a cell wall and their resistance to detergent lysis is active, requiring continued energy metabolism, and may be assisted by a complete cessation of endocytosis, as measured by uptake of the dye FM1-43. Microarray analysis shows that aspidocytes have a distinct pattern of gene expression, with a number of genes up-regulated that are predicted to be involved in lipid metabolism. Aspidocytes were initially detected in a hypersensitive mutant, in which the AMP deaminase gene is disrupted, suggesting that the inductive pathway involves AMP levels or metabolism. Since aspidocytes can also be induced from wild-type cells and are much more resistant than amoebae to a membrane-disrupting antibiotic, it is possible that they are an adaptation allowing cells to survive a sudden onslaught of toxins in the wild.

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2007-02-01
2019-10-21
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Aspidocyte formation. Starving cells of strain HM1158 were induced to form aspidocytes in the standard conditions (see Methods of main paper) by 25 µM Cd and 100 nM DIF-1, except that they were plated on coverslips in a chambered microscope slide. Filming started after 5 h of incubation and covers a period of 50 min (1 frame/30 s). [ .mov file] (2800 kb) Aspidocyte dissolution. Aspidocytes were induced from starving cells of strain HM1158 by 25 µM Cd and 100 nM DIF-1 overnight in the standard conditions. They were then washed free of inducers and plated on a coverslip for filming, which started immediately. The film covers a period of 20 min (1 frame per 20 s). [ .mov file] (3520 kb) Genes significantly up-regulated in aspidocytes induced by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (106 kb) Genes significantly down-regulated in aspidocytes by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (119 kb) A Gene identity at dictyBase ( http://dictybase.org) B. Chromosome C. Presence of signal peptide D. Number of transmembrane domains E. Presence of GPI anchor F. Pfam domain G. Additional annotation H. Induction ratio (log ) with Cd /DES compared to un-induced control I. Induction ratio (log ) with thiabendazole compared to un-induced control J. Adjusted probability for Cd /DES induction K. Adjusted probability for thiabendazole induction

MOVIE

Aspidocyte formation. Starving cells of strain HM1158 were induced to form aspidocytes in the standard conditions (see Methods of main paper) by 25 µM Cd and 100 nM DIF-1, except that they were plated on coverslips in a chambered microscope slide. Filming started after 5 h of incubation and covers a period of 50 min (1 frame/30 s). [ .mov file] (2800 kb) Aspidocyte dissolution. Aspidocytes were induced from starving cells of strain HM1158 by 25 µM Cd and 100 nM DIF-1 overnight in the standard conditions. They were then washed free of inducers and plated on a coverslip for filming, which started immediately. The film covers a period of 20 min (1 frame per 20 s). [ .mov file] (3520 kb) Genes significantly up-regulated in aspidocytes induced by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (106 kb) Genes significantly down-regulated in aspidocytes by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (119 kb) A Gene identity at dictyBase ( http://dictybase.org) B. Chromosome C. Presence of signal peptide D. Number of transmembrane domains E. Presence of GPI anchor F. Pfam domain G. Additional annotation H. Induction ratio (log ) with Cd /DES compared to un-induced control I. Induction ratio (log ) with thiabendazole compared to un-induced control J. Adjusted probability for Cd /DES induction K. Adjusted probability for thiabendazole induction

MOVIE

Aspidocyte formation. Starving cells of strain HM1158 were induced to form aspidocytes in the standard conditions (see Methods of main paper) by 25 µM Cd and 100 nM DIF-1, except that they were plated on coverslips in a chambered microscope slide. Filming started after 5 h of incubation and covers a period of 50 min (1 frame/30 s). [ .mov file] (2800 kb) Aspidocyte dissolution. Aspidocytes were induced from starving cells of strain HM1158 by 25 µM Cd and 100 nM DIF-1 overnight in the standard conditions. They were then washed free of inducers and plated on a coverslip for filming, which started immediately. The film covers a period of 20 min (1 frame per 20 s). [ .mov file] (3520 kb) Genes significantly up-regulated in aspidocytes induced by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (106 kb) Genes significantly down-regulated in aspidocytes by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (119 kb) A Gene identity at dictyBase ( http://dictybase.org) B. Chromosome C. Presence of signal peptide D. Number of transmembrane domains E. Presence of GPI anchor F. Pfam domain G. Additional annotation H. Induction ratio (log ) with Cd /DES compared to un-induced control I. Induction ratio (log ) with thiabendazole compared to un-induced control J. Adjusted probability for Cd /DES induction K. Adjusted probability for thiabendazole induction

EXCEL

Aspidocyte formation. Starving cells of strain HM1158 were induced to form aspidocytes in the standard conditions (see Methods of main paper) by 25 µM Cd and 100 nM DIF-1, except that they were plated on coverslips in a chambered microscope slide. Filming started after 5 h of incubation and covers a period of 50 min (1 frame/30 s). [ .mov file] (2800 kb) Aspidocyte dissolution. Aspidocytes were induced from starving cells of strain HM1158 by 25 µM Cd and 100 nM DIF-1 overnight in the standard conditions. They were then washed free of inducers and plated on a coverslip for filming, which started immediately. The film covers a period of 20 min (1 frame per 20 s). [ .mov file] (3520 kb) Genes significantly up-regulated in aspidocytes induced by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (106 kb) Genes significantly down-regulated in aspidocytes by Cd /DES or thiabendazole (or in both conditions). [ .xls file] (119 kb) A Gene identity at dictyBase ( http://dictybase.org) B. Chromosome C. Presence of signal peptide D. Number of transmembrane domains E. Presence of GPI anchor F. Pfam domain G. Additional annotation H. Induction ratio (log ) with Cd /DES compared to un-induced control I. Induction ratio (log ) with thiabendazole compared to un-induced control J. Adjusted probability for Cd /DES induction K. Adjusted probability for thiabendazole induction

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