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Journal of Histochemistry and Cytochemistry, Vol. 47, 1201-1212, September 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Cerium-based Histochemical Demonstration of Oxidative Stress in Taurocholate-induced Acute Pancreatitis in Rats: A Confocal Laser Scanning Microscopic Study

Géza Teleka, Jean-Yves Scoazecb, Jacques Chariota, Robert Ducroca, Gérard Feldmannb, and Claude Rozéa
a INSERM U410, Faculté de Médecine Xavier Bichat, Université Paris 7, Paris, France
b INSERM U327, Faculté de Médecine Xavier Bichat, Université Paris 7, Paris, France

Correspondence to: Claude Rozé, INSERM U410, Faculté de Médecine Xavier Bichat, 16 Rue Henri Huchard, BP 416, 75870 Paris, France.

Direct in vivo histological detection of oxygen-derived free radicals (OFRs) in inflammatory conditions is not fully resolved. We report an application of cerium histochemistry (in which capture of OFRs by Ce atoms results in laser-reflectant cerium-perhydroxide precipitates) combined with reflectance confocal laser scanning microscopy (CLSM) to demonstrate the evolution of oxidative stress in taurocholate-induced acute pancreatitis (AP) in rats. Animals were perfused with CeCl3 in vivo and cryostat sections of pancreata were studied by CLSM. Vascular endothelium was immunolabeled for PECAM-1. OFR production by isolated polymorphonuclear leukocytes (PMNs) incubated in vitro with CeCl3 was quantified by image analysis. In the pancreas, strong OFR-derived cerium reflectance signals were seen in acinar cells at 1–2 hr, capillaries and small venules were frequently engorged by cerium precipitates, and adherent PMNs presented weak intracellular reflectance signals. At 8–24 hr, acinar cell OFR production decreased, whereas adherent/transmigrated PMNs displayed abundant intra- and pericellular reflectance. PECAM-1 expression was unchanged. PMNs from ascites or blood showed significant (p<0.01) time-dependent OFR production, plateauing from 2 hr. The modified cerium capture/CLSM method allows the co-demonstration of in vivo oxidative stress and cellular structures labeled with fluorescent markers. In vivo oxidative stress was shown histologically for the first time in experimental AP. (J Histochem Cytochem 47:1201–1212, 1999)

Key Words: experimental acute pancreatitis, reactive oxygen species, cerium histochemistry, reflectance confocal, microscopy, polymorphonuclear leukocytes, PECAM, immunofluorescence


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