spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

First published online November 4, 2005
Journal of Experimental Biology 208, 4345-4354 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01897
This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ares, I. R.
Right arrow Articles by Botana, L. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ares, I. R.
Right arrow Articles by Botana, L. M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Actin cytoskeleton of rabbit intestinal cells is a target for potent marine phycotoxins

I. R. Ares1, M. C. Louzao1, M. R. Vieytes2, T. Yasumoto3 and L. M. Botana1,*

1 Departamento de Farmacología, Facultad de Veterinaria de Lugo, Universidad de Santiago de Compostela, 27002 Lugo, Spain
2 Departamento de Fisiología Animal, Facultad de Veterinaria de Lugo, Universidad de Santiago de Compostela, 27002 Lugo, Spain
3 Japan Food Research Laboratories, Tama, Tokyo 206-0025, Japan



View larger version (43K):

[in a new window]
  		Fig. 1. Analysis of F-actin level in enterocytes by using laser-scanning cytometry. Typical experiment showing the effect on enterocytes of a 4 h incubation with (A) 1 µmol l–1 PTX-6 or (B) 1 µmol l–1 YTX. PTX-6 provokes a high loss of fluorescence intensity in cells compared to controls (A), while almost no effect is observed with YTX (B).

 


View larger version (16K):

[in a new window]
  		Fig. 2. Effect of all toxins studied on F-actin cytoskeleton. Data obtained using laser scanner cytometry are expressed as percentage fluorescence of enterocytes incubated with toxins compared to controls (100%). Values are means ± S.E.M.

 


View larger version (55K):

[in a new window]
  		Fig. 3. Control cells (A,D) and cells incubated with 1 µmol l–1 PTX-6 (B) or 5 nmol l–1 MTX (E) for 4 h. Confocal microscopy showed that both marine toxins affect the microfilament network without modifying the morphological pattern of isolated intestinal cells. (C,D) Transmission images of cells incubated with the toxins.

 


View larger version (53K):

[in a new window]
  		Fig. 4. Analysis of F-actin levels in enterocytes using laser-scanning cytometry. Typical experiment showing the effect on enterocytes of a 4 h incubation with (A) 4 nmol l–1 CTX-3C or (B) 5 nmol l–1 MTX. MTX provokes a high loss of fluorescence intensity in cells compared to controls (A) while scarcely any effect is observed with CTX-3C (B).

 


View larger version (42K):

[in a new window]
  		Fig. 5. Analysis of F-actin levels in enterocytes using laser-scanning cytometry. Representative experiment of the effect of (A) 75 nmol l–1 palytoxin or (B) 75 nmol l–1 ostreocin-D incubated for 4 h with intestinal cells. In this case, both palytoxin (A) and ostreocin-D (B) cause an important reduction in emitted fluorescence by treated cells compared to controls.

 


View larger version (53K):

[in a new window]
  		Fig. 6. Control cells (A,D) and cells incubated with 75 nmol l–1 palytoxin (B) or 75 nmol l–1 ostreocin-D (E) for 4 h. Confocal microscopy shows that both toxins modify actin filaments, but there was no change in shape of the intestinal cells. (C,F) Transmission images of cells exposed to the toxins.

 


View larger version (47K):

[in a new window]
  		Fig. 7. Analysis of F-actin level in enterocytes using laser-scanning cytometry. Representative experiment of the effect of (A) 250 nmol l–1 Pbtx-3 or (B) 20 nmol l–1 Pbtx-9 incubated for 4 h with intestinal cells. There is almost no change in distribution of fluorescence between cells incubated with the Pbtx-3 (A) and Pbtx-9 (B) and the controls.

 


View larger version (60K):

[in a new window]
  		Fig. 8. Analysis of F-actin level in enterocytes using laser-scanning cytometry. Representative experiment of the effect of (A) 5 nmol l–1 MTX, (B) 75 nmol l–1 palytoxin or (C) 75 nmol l–1 ostreocin-D incubated for 4 h with intestinal cells in a Ca2+-free medium. MTX (A), palytoxin (B) and ostreocin-D (C) evoked a diminution in fluorescence intensity of treated cells, but this was less marked than when Ca2+ was present (see Fig. 9).

 


View larger version (17K):

[in a new window]
  		Fig. 9. Comparison of F-actin decrease in intestinal cells treated with MTX, palytoxin and ostreocin-D in a medium with or without Ca2+. Values (means ± S.E.M.) are indicated as a percentage of fluorescence of cells incubated with toxins with compared to controls (100%).

 

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?



© The Company of Biologists Ltd 2005