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Localization of Satratoxin-G in Stachybotrys chartarum Spores and Spore-Impacted Mouse Lung Using Immunocytochemistry

Department of Biology Saint Mary's University, Halifax, Nova Scotia, B3H 3C3

James J. Pestka

Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan, USA

Dorr G. Dearborn

Department of Pediatrics, Case Western University, Cleveland, Ohio, USA

Thomas G. Rand

Department of Biology Saint Mary's University, Halifax, Nova Scotia, B3H 3C3, thomas.rand{at}STMARYS.CA

Satratoxin-G (SG) is the major macrocyclic trichothecene mycotoxin produced by Stachybotrys chartarum (atra) and has been implicated as a cause of a number of animal and human health problems including pulmonary hemorrhage in infants. However, there is little understanding where this toxin is localized in the spores and mycelial fragments of this species or in the lung impacted by SG-sequestered spores. The purpose of this study was to evaluate the distribution of SG in S. chartarum spores and mycelium in culture, and spore-impacted mouse lung in vivo, using immunocytochemistry. SG was localized predominately in S. chartarum spores with moderate labelling of the phialide-apex walls. Labelling was primarily along the outer plasmalemma surface and in the inner wall layer. Only modest labelling was observed in hyphae. Toxin localization at these sites supports the position that spores contain the highest satratoxin concentrations and that the toxin is constitutively produced. In impacted mouse lung, highest SG labelling was detected in lysosomes, along the inside of the nuclear membrane in nuclear heterochromatin and RER within alveolar macrophages. Alveolar type II cells also showed modest labelling of the nuclear heterochromatin and RER. There was no evidence that the toxin accumulated in the neutrophils, fibroblasts, or other cells associated with the granulomas surrounding spores or mycelial fragments. These observations indicate that SG displays a high degree of cellular specificity with respect to its uptake in mouse lung. They further indicate that the alveolar macrophages play an important role in the sequestration and immobilization of low concentrations of the toxin.

Key Words: Stachybotrys chartarum • conidia • trichothecenes • satratoxin-G • intratracheal instillation • immunocytochemistry • ultrastructure.

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