Immunohistochemical localization of androgen receptor in rat caput epididymis during postnatal development

Abstract

Objectives: The aim of this study was to investigate the developmental pattern of androgen receptor (AR) in caput epididymis. Materials and methods: In this study three randomly selected rats were sacrificed at ages 21, 56, 90 and 120 days old. All male rats were anesthetized with ethyl ether before killing. Then, the caput epididymides were removed and fixed in Bouin's fixative at +4°C for 36 hour. Afterwards the tissue samples were embedded in paraffin for routine histological methods. Later the tissues were sectioned at 5μm and mounted on poly-L-lysin-coated slides. To solve the antigen masking problem, we performed microwave stimulated antigen retrieval technique before the immunohistochemical staining. Avidin-Biotin-Peroxidase Complex (ABC) method was applied for immunohistochemical staining. Results: In all age groups of rats studied, positive immunohistochemical staining for the AR appeared in nuclei of epididymal cells. The staining intensity of AR positive cells did not change depending on age. In caput epididymis, immunostainable AR was found in tubular epithelial cells (principal cells, basal cells and apical cells) and peritubular smooth muscle cells. The AR staining in the epithelial cells appeared to be stronger than in the peritubular smooth muscle cells. In the epithelial cells; staining intensity was stronger in principal cells than in basal cells and apical cells. Conclusion: Staining intensity of AR positive epididymal cells irrespective of age indicated the necessity of androgens for postnatal differentiation and maintaining the structure of the epididymis. Stronger staining intensity in principal cells suggested that principal cells are more sensitive to androgen stimulation.

Preclinical immunohistochemical biomarkers for testicular changes in hyperlipidemic rats and the role of Ocimum gratissimum

“…In a study conducted on the peritubular myoid cells, it was reported that these cells contain a large amount of ARs, and the depletion of the receptors in these cells caused a decrease in testis size and oligospermia [27]. The present work showed that AR expression in myoid cells was decreased after 2 weeks of tyloxapol injection and markedly increased in rats supplemented with O. gratissimum.…”

“…The testes were dissected out carefully from each animal without damaging the tunica albuginea and were fixed in Bouin's fixative for 24 h, processed for light microscopy to obtain 5-µm-thick paraffin sections, and stained with H&E to verify histological details [25]; immunohistochemical staining for detection of AR and PCNA was also carried out [26,27].…”

Androgen receptor gene mutations are associated with male infertility in Northeast China: Clinical features and identification of two novel mutations

“…AR has been reported to be a steroid receptor in the nuclear receptor superfamily and is mainly localised in the nucleus of the following cells: sertoli cells, peritubular myoid cells and periarterial cells, Leydig cells and fibroblasts (Gur & Timurkaan, ; Suárez‐Quian, Martínez‐García, Nistal, & Regadera, ; Timurkaan, Gur, & Karan, ; Van Roijen et al, ). Studies using mouse AR gene knockout models found that AR gene defects can cause insufficient androgen action, leading to spermatogenic dysfunction by blocking the meiosis process of spermatogenesis (Chang et al, ; De Gendt et al, ).…”

Thiamine deficiency and oxidative stress induced by prolonged metronidazole therapy can explain its side effects of neurotoxicity and infertility in experimental animals: Effect of grapefruit co-therapy

“…Then, these sections were stained with haematoxylin and eosin (H&E), 22 for examination by light microscopy. Additionally, testicle and epididymis samples were further examined by Masson's trichrome staining 23 for the collagenous fibres, Periodic acid-Schiff (PAS) staining 24 for general carbohydrates and immunohistochemical examination for the presence of androgen receptor detection 25 using a polyclonal antibody (catalogue number #RB-9030-R7, Thermo Fisher Scientific, Fremont, California, USA). For immunohistochemistry, 5-mm-thick sections were mounted on poly-Llysine-coated slides and heated at 60 C for 1 h in an oven to enhance adherence to the slide.…”