Covalent affinity labeling, radioautography, and immunocytochemistry localize the glucocorticoid receptor in rat testicular Leydig cells

Journal Article · Fri Dec 01 04:00:00 EST 1989 · American Journal of Anatomy; (USA)

DOI:https://doi.org/10.1002/aja.1001860406· OSTI ID:5085512

Stalker, A; Hermo, L; Antakly, T ^[1]

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1. McGill Univ., Montreal, Quebec (Canada)

The presence and distribution of glucocorticoid receptors in the rat testis were examined by using 2 approaches: in vivo quantitative radioautography and immunocytochemistry. Radioautographic localization was made possible through the availability of a glucocorticoid receptor affinity label, dexamethasone 21-mesylate, which binds covalently to the glucocorticoid receptor, thereby preventing dissociation of the steroid-receptor complex. Adrenalectomized adult rats were injected with a tritiated (3H) form of this steroid into the testis and the tissue was processed for light-microscope radioautography. Silver grains were observed primarily over the Leydig cells of the interstitial space and to a lesser extent, over the cellular layers which make up the seminiferous epithelium, with no one cell type showing preferential labeling. To determine the specificity of the labeling, a 25- or 50-fold excess of unlabeled dexamethasone was injected simultaneously with the same dose of (3H)-dexamethasone 21-mesylate. In these control experiments, a marked reduction in label intensity was noted over the Leydig as well as tubular cells. Endocytic macrophages of the interstitium were non-specifically labeled, indicating uptake of the ligand possibly by fluid-phase endocytosis. A quantitative analysis of the label confirmed the presence of statistically significant numbers of specific binding sites for glucocorticoids in both Leydig cells and the cellular layers of the seminiferous epithelium; 86% of the label was found over Leydig cells, and only 14% over the cells of the seminiferous epithelium. These binding data were confirmed by light-microscope immunocytochemistry using a monoclonal antibody to the glucocorticoid receptor.

• OSTI ID: 5085512
• Journal Information: American Journal of Anatomy; (USA), Journal Name: American Journal of Anatomy; (USA) Vol. 186:4; ISSN AJANA; ISSN 0002-9106
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADRENAL HORMONES
AFFINITY
ANIMAL CELLS
ANIMAL TISSUES
ANIMALS
ANTIBODIES
AUTORADIOGRAPHY
BIOCHEMICAL REACTION KINETICS
BODY
CONNECTIVE TISSUE CELLS
CORTICOSTEROIDS
DEXAMETHASONE
DISTRIBUTION
EPITHELIUM
GLUCOCORTICOIDS
GONADS
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
KETONES
KINETICS
LABELLING
MACROPHAGES
MALE GENITALS
MAMMALS
MEMBRANE PROTEINS
MONOCLONAL ANTIBODIES
ORGANIC COMPOUNDS
ORGANS
PHAGOCYTES
PREGNANES
PROTEINS
RATS
REACTION KINETICS
RECEPTORS
RODENTS
SOMATIC CELLS
STEROIDS
SUBCELLULAR DISTRIBUTION
TESTES
TISSUES
TRITIUM COMPOUNDS
VERTEBRATES