Development of in vitro models for a better understanding of the early pathogenesis of Batrachochytrium dendrobatidis infections in amphibians
- Author
- Pascale Van Rooij (UGent) , An Martel (UGent) , Melanie Brutyn (UGent) , Sofie Maes (UGent) , Koen Chiers (UGent) , Lieven Van Waeyenberghe (UGent) , Siska Croubels (UGent) , Freddy Haesebrouck (UGent) and Frank Pasmans (UGent)
- Organization
- Abstract
- Batrachochytrium dendrobatidis, the causal agent of chytridiomycosis, is implicated in the global decline of amphibians. This chytrid fungus invades keratinised epithelial cells, and infection is mainly associated with epidermal hyperplasia and hyperkeratosis. Since little is known about the pathogenesis of chytridiomycosis, this study was designed to optimise the conditions under which primary keratinocytes and epidermal explants of amphibian skin could be maintained ex vivo for several days. The usefulness of the following set-ups for pathogenesis studies was investigated: a) cultures of primary keratinocytes; b) stripped epidermal (SE) explants; c) full-thickness epidermal (FTE) explants on Matrigel (TM); d) FTE explants in cell culture inserts; and e) FTE explants in Ussing chambers. SE explants proved most suitable for short-term studies, since adherence of fluorescently-labelled zoospores to the superficial epidermis could be observed within one hour of infection. FTE explants in an Ussing chamber set-up are most suitable for the study of the later developmental stages of B. dendrobatidis in amphibian skin up to five days post-infection. These models provide a good alternative for in vivo experiments, and reduce the number of experimental animals needed.
- Keywords
- Batrachochytrium dendrobatidis, primary keratinocytes, epidermal explants, in vitro model, chytridiomycosis, amphibian skin, FROGS LITORIA-CAERULEA, POPULATION DECLINES, CUTANEOUS CHYTRIDIOMYCOSIS, CHYTRID FUNGUS, XENOPUS-LAEVIS, DISEASE, DEFENSES, TIME
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-1113117
- MLA
- Van Rooij, Pascale, et al. “Development of in Vitro Models for a Better Understanding of the Early Pathogenesis of Batrachochytrium Dendrobatidis Infections in Amphibians.” ATLA-ALTERNATIVES TO LABORATORY ANIMALS, vol. 38, no. 6, 2010, pp. 519–28.
- APA
- Van Rooij, P., Martel, A., Brutyn, M., Maes, S., Chiers, K., Van Waeyenberghe, L., … Pasmans, F. (2010). Development of in vitro models for a better understanding of the early pathogenesis of Batrachochytrium dendrobatidis infections in amphibians. ATLA-ALTERNATIVES TO LABORATORY ANIMALS, 38(6), 519–528.
- Chicago author-date
- Van Rooij, Pascale, An Martel, Melanie Brutyn, Sofie Maes, Koen Chiers, Lieven Van Waeyenberghe, Siska Croubels, Freddy Haesebrouck, and Frank Pasmans. 2010. “Development of in Vitro Models for a Better Understanding of the Early Pathogenesis of Batrachochytrium Dendrobatidis Infections in Amphibians.” ATLA-ALTERNATIVES TO LABORATORY ANIMALS 38 (6): 519–28.
- Chicago author-date (all authors)
- Van Rooij, Pascale, An Martel, Melanie Brutyn, Sofie Maes, Koen Chiers, Lieven Van Waeyenberghe, Siska Croubels, Freddy Haesebrouck, and Frank Pasmans. 2010. “Development of in Vitro Models for a Better Understanding of the Early Pathogenesis of Batrachochytrium Dendrobatidis Infections in Amphibians.” ATLA-ALTERNATIVES TO LABORATORY ANIMALS 38 (6): 519–528.
- Vancouver
- 1.Van Rooij P, Martel A, Brutyn M, Maes S, Chiers K, Van Waeyenberghe L, et al. Development of in vitro models for a better understanding of the early pathogenesis of Batrachochytrium dendrobatidis infections in amphibians. ATLA-ALTERNATIVES TO LABORATORY ANIMALS. 2010;38(6):519–28.
- IEEE
- [1]P. Van Rooij et al., “Development of in vitro models for a better understanding of the early pathogenesis of Batrachochytrium dendrobatidis infections in amphibians,” ATLA-ALTERNATIVES TO LABORATORY ANIMALS, vol. 38, no. 6, pp. 519–528, 2010.
@article{1113117,
abstract = {{Batrachochytrium dendrobatidis, the causal agent of chytridiomycosis, is implicated in the global decline of amphibians. This chytrid fungus invades keratinised epithelial cells, and infection is mainly associated with epidermal hyperplasia and hyperkeratosis. Since little is known about the pathogenesis of chytridiomycosis, this study was designed to optimise the conditions under which primary keratinocytes and epidermal explants of amphibian skin could be maintained ex vivo for several days. The usefulness of the following set-ups for pathogenesis studies was investigated: a) cultures of primary keratinocytes; b) stripped epidermal (SE) explants; c) full-thickness epidermal (FTE) explants on Matrigel (TM); d) FTE explants in cell culture inserts; and e) FTE explants in Ussing chambers. SE explants proved most suitable for short-term studies, since adherence of fluorescently-labelled zoospores to the superficial epidermis could be observed within one hour of infection. FTE explants in an Ussing chamber set-up are most suitable for the study of the later developmental stages of B. dendrobatidis in amphibian skin up to five days post-infection. These models provide a good alternative for in vivo experiments, and reduce the number of experimental animals needed.}},
author = {{Van Rooij, Pascale and Martel, An and Brutyn, Melanie and Maes, Sofie and Chiers, Koen and Van Waeyenberghe, Lieven and Croubels, Siska and Haesebrouck, Freddy and Pasmans, Frank}},
issn = {{0261-1929}},
journal = {{ATLA-ALTERNATIVES TO LABORATORY ANIMALS}},
keywords = {{Batrachochytrium dendrobatidis,primary keratinocytes,epidermal explants,in vitro model,chytridiomycosis,amphibian skin,FROGS LITORIA-CAERULEA,POPULATION DECLINES,CUTANEOUS CHYTRIDIOMYCOSIS,CHYTRID FUNGUS,XENOPUS-LAEVIS,DISEASE,DEFENSES,TIME}},
language = {{eng}},
number = {{6}},
pages = {{519--528}},
title = {{Development of in vitro models for a better understanding of the early pathogenesis of Batrachochytrium dendrobatidis infections in amphibians}},
volume = {{38}},
year = {{2010}},
}