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

Journal Article · Fri Dec 01 00:00:00 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), Vol. 186:4; ISSN 0002-9106
• Country of Publication: United States
• Language: English

Related Subjects

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