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Journal of Histochemistry and Cytochemistry, Vol. 50, 21-32, January 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

CL316,243 and Cold Stress Induce Heterogeneous Expression of UCP1 mRNA and Protein in Rodent Brown Adipocytes

Saverio Cintia, Raffaella Cancelloa, Maria Cristina Zingarettia, Enzo Ceresia, Rita De Matteisa, Antonio Giordanoa, Jean Himms–Hagenb, and Daniel Ricquierc
a Institute of Normal Human Morphology, Faculty of Medicine, University of Ancona, Italy
b Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Canada
c Centre de Recherche sur l'Endocrinologie Moléculaire et le Développement, Centre National de la Recherche Scientifique, Meudon, France

Correspondence to: Saverio Cinti, Inst. of Normal Human Morphology, Faculty of Medicine, University of Ancona, Via Tronto 10/A, 60020 Torrette–Ancona, Italy. E-mail: cinti@popcsi.unian.it

Uncoupling protein 1 (UCP1), the mammalian thermogenic mitochondrial protein, is found only in brown adipocytes, but its expression by immunohistochemistry is not homogeneous. Here we present evidence that the non-homogeneous pattern of immunostaining for UCP1 (referred to as the "Harlequin phenomenon") is particularly evident after acute and chronic cold (4C) stimulus and after administration of a specific ß3-adrenoceptor agonist (CL316,243). Accordingly, mRNA in situ expression confirmed the UCP1 non-homogeneous pattern of gene activation under conditions of adrenergic stimulus. Furthermore, morphometric analysis of immunogold-stained thin sections showed that UCP1–gold particle density was different among neighboring brown adipocytes with mitochondria of the same size and density. When the adrenergic stimulus was reduced in warm-acclimated animals (28C), UCP1 protein and mRNA expression was reduced and consequently the Harlequin phenomenon was barely visible. These data suggest the existence of an alternative and controlled functional recruitment of brown adipocytes in acute adrenergically stressed animals, possibly to avoid heat and metabolic damage in thermogenically active cells. Of note, the heat shock protein heme oxygenase 1 (HO1) is heterogeneously expressed in adrenergically stimulated brown adipose tissue and, specifically, cells expressing strong immunoreactivity for UCP1 also strongly express HO1.

(J Histochem Cytochem 50:21–31, 2002)

Key Words: uncoupling protein 1, ß3-adrenoceptor agonist, brown adipose tissue, thermogenesis, heme oxygenase, heat shock protein


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