Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining

In the field of studies of acute toxicity induced by bacterial agents, Shiga toxins have been relevant due to the severity of the extra-intestinal diseases they cause. Numerous studies have shown that Shiga toxin induced apoptosis in different cell types; however, this important process has been lit...

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Autores principales:	Alvarez,M, Urbina,G, Perdomo,L
Lenguaje:	English
Publicado:	Sociedad Chilena de Anatomía 2014
Materias:	Zebrafish (Danio rerio) Shigella dysenteriae Apoptosis Acridine Orange Ethidium Bromide
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022014000100014
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spelling	oai:scielo:S0717-950220140001000142015-11-13Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo StainingAlvarez,MUrbina,GPerdomo,L Zebrafish (Danio rerio) Shigella dysenteriae Apoptosis Acridine Orange Ethidium Bromide In the field of studies of acute toxicity induced by bacterial agents, Shiga toxins have been relevant due to the severity of the extra-intestinal diseases they cause. Numerous studies have shown that Shiga toxin induced apoptosis in different cell types; however, this important process has been little studied in vivo experimental models. In this study, the effects of excretion products of Shigella dysenteriae, in which Shiga toxin is present, were investigated on early larval stages of Zebrafish, an animal model with many advantages over other in vivo experimental models traditionally used. Both the collection of eggs and larvae of Zebrafish, and the product from excretion from Shigella dysenteriae (SdyEP) were performed according to laboratory standards. Also, toxicity bioassay, larvae treatment with pure and diluted solution, 10-1, 10-2, 10-3 , 10-4 and 10-5, v/v SdyEP and cell death in vivo using Acridine Orange (AO) and Ethidium Bromide (EB) were applied. The excretion product of Shigella dysenteriae (SdyEP) effect was expressed in terms of larval mortality and dependent dilution rather than incubation time. The larval population surviving treatment with Shigella excretion product presents severe morphological effects. The larval population generally presents notable severe morphological damage, the necrosis state is represented by the opacity of the larvae after being treated for 24 h (b) compared to control. Other changes associated with larval anatomy were also observed; particularly the caudal end curvature was significant into 10%. The use of AO/EB revealed a distribution pattern from fluorescence into green and orange in surviving larvae SdyEP poisoning, there was a large population of dead cells around the anal and caudal region as evidenced by the presence of orange nuclei in greater numbers as controls in the larvae. The results support the application of coloring AO/EB in Zebrafish experimental models for the evaluation of the toxic action of new molecules and new products with therapeutic potential.info:eu-repo/semantics/openAccessSociedad Chilena de AnatomíaInternational Journal of Morphology v.32 n.1 20142014-03-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022014000100014en10.4067/S0717-95022014000100014
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Zebrafish (Danio rerio) Shigella dysenteriae Apoptosis Acridine Orange Ethidium Bromide
spellingShingle	Zebrafish (Danio rerio) Shigella dysenteriae Apoptosis Acridine Orange Ethidium Bromide Alvarez,M Urbina,G Perdomo,L Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining
description	In the field of studies of acute toxicity induced by bacterial agents, Shiga toxins have been relevant due to the severity of the extra-intestinal diseases they cause. Numerous studies have shown that Shiga toxin induced apoptosis in different cell types; however, this important process has been little studied in vivo experimental models. In this study, the effects of excretion products of Shigella dysenteriae, in which Shiga toxin is present, were investigated on early larval stages of Zebrafish, an animal model with many advantages over other in vivo experimental models traditionally used. Both the collection of eggs and larvae of Zebrafish, and the product from excretion from Shigella dysenteriae (SdyEP) were performed according to laboratory standards. Also, toxicity bioassay, larvae treatment with pure and diluted solution, 10-1, 10-2, 10-3 , 10-4 and 10-5, v/v SdyEP and cell death in vivo using Acridine Orange (AO) and Ethidium Bromide (EB) were applied. The excretion product of Shigella dysenteriae (SdyEP) effect was expressed in terms of larval mortality and dependent dilution rather than incubation time. The larval population surviving treatment with Shigella excretion product presents severe morphological effects. The larval population generally presents notable severe morphological damage, the necrosis state is represented by the opacity of the larvae after being treated for 24 h (b) compared to control. Other changes associated with larval anatomy were also observed; particularly the caudal end curvature was significant into 10%. The use of AO/EB revealed a distribution pattern from fluorescence into green and orange in surviving larvae SdyEP poisoning, there was a large population of dead cells around the anal and caudal region as evidenced by the presence of orange nuclei in greater numbers as controls in the larvae. The results support the application of coloring AO/EB in Zebrafish experimental models for the evaluation of the toxic action of new molecules and new products with therapeutic potential.
author	Alvarez,M Urbina,G Perdomo,L
author_facet	Alvarez,M Urbina,G Perdomo,L
author_sort	Alvarez,M
title	Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining
title_short	Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining
title_full	Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining
title_fullStr	Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining
title_full_unstemmed	Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining
title_sort	excretion product of shigella dysenteriae (sdyep) induced cell death in early larval stage of zebrafish (danio rerio): acridine orange and ethidium bromide (ao/eb) in vivo staining
publisher	Sociedad Chilena de Anatomía
publishDate	2014
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022014000100014
work_keys_str_mv	AT alvarezm excretionproductofshigelladysenteriaesdyepinducedcelldeathinearlylarvalstageofzebrafishdaniorerioacridineorangeandethidiumbromideaoebinvivostaining AT urbinag excretionproductofshigelladysenteriaesdyepinducedcelldeathinearlylarvalstageofzebrafishdaniorerioacridineorangeandethidiumbromideaoebinvivostaining AT perdomol excretionproductofshigelladysenteriaesdyepinducedcelldeathinearlylarvalstageofzebrafishdaniorerioacridineorangeandethidiumbromideaoebinvivostaining
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Excretion Product of Shigella dysenteriae (SdyEP) Induced Cell Death in Early Larval Stage of Zebrafish (Danio rerio): Acridine Orange and Ethidium Bromide (AO/EB) in vivo Staining

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