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Localization of Inducible Nitric Oxide Synthase In a Rat Model of Silicosis

Published online by Cambridge University Press:  02 July 2020

L. Millecchia
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV26505
P. Willard
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV26505
A. Hubbs
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV26505
D. Porter
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV26505
V. Castranova
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV26505
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Extract

We are participating in a collaborative project at NIOSH on mechanisms of silicosis, in which rats were exposed to filtered air (control) or silica aerosol of 15 mg/m3 (6 hr/day, 5 days/week), with assays conducted at intervals over a 6 month period. We are especially interested in the role of nitric oxide (NO) in the pathophysiology of lung disease. One source of NO is the alveolar macrophage, known to express the inducible form of nitric oxide synthase (iNOS); we used immunohistochemical techniques to determine whether iNOS is upregulated in alveolar macrophages and other lung sources in response to silica.

Silica-exposed and air control animals were sacrificed after 10, 20, 41, 79, and 116 days of exposure. Formalin fixed, paraffin embedded tissues were cut at 5μm, deparaffinized in xylene and rehydrated. Slides were microwaved in citrate buffer, pH 6.0 for antigen retrieval.2 After blocking endogenous peroxidase in 3% H2O2:Methanol (1:1), slides were placed in 10% BSA for 30 min, then incubated overnight in the primary antibody (Monoclonal anti-iNOS, Transduction Laboratories, N32020, 1:50 dilution)

Type
Pathology
Copyright
Copyright © Microscopy Society of America

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References

1.Porter, D.W., et al., (1999) The Toxicologist 43:In press.Google Scholar
2.Takahashi, A., et al., (1997) Cancer Res. 57:1233Google Scholar
3.Suarez, F., et al., (1999) The Toxicologist 43: In press.Google Scholar