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Isolate Cell-Type-Specific RNAs from Snap-Frozen Heterogeneous Tissue Samples without Cell Sorting
Published on: December 8, 2021
Expression of the glucocorticoid receptor and 11 beta-hydroxysteroid dehydrogenase 2 in pig testes cells along fetal
1Institut für Tierhaltung und Tierzüchtung, Universität Hohenheim, Garbenstr. 17, 70599 Stuttgart, Germany.
Abstract:
The glucocorticoid (GC)-cortisol receptor (GCR)-11 beta-hydroxysteroid dehydrogenase 2 (11 beta-HSD2) system is involved in the regulation of Leydig cell function and spermatogenesis in mature animals. Herein, we describe the expression of the GCR and 11 beta-HSD2 and the occurrence of apoptosis during fetal development. Male fetuses were collected from Weeks 6, 10, 13, and 15 of pregnancy and from neonates. The testes were used for the immunocytochemical staining of GCR, 11 beta-HSD2 and for terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) staining of apoptosis. Apoptosis did not occur in any Leydig cells, but approximately 30% expressed GCR and 11 beta-HSD2. The number of GCR-positive cells was similar at all stages, but the number of 11 beta-HSD2-positive cells tended to be higher at Weeks 6 and 15. Steroid synthesis was also higher compared with Weeks 10 and 13. Apoptosis occurred in only a few germ cells. Nearly all germ cells were GCR positive at Weeks 10 and 13, when 11 beta-HSD2 was also increased. The total number of 11 beta-HSD2-positive germ cells was approximately 30%. Thus, elevated GCR expression coincided with the differentiation of gonocytes to spermatogonia and their migration to the basal lamina.
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