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.

Keywords

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

Öz

Amaç: Bu çalışmanın amacı postnatal süreçte kaput epididimisteki androjen reseptör (AR)'ün gelişimsel dağılımını araştırmaktır. Gereç ve yöntem: Bu çalışmada; rastgele seçilen üçer adet sıçan 21, 56, 90 ve 120 günlük yaşlarda dietil eter ile uyutuldu. Bunu takiben kaput epididimisler, hızlı bir şekilde vücuttan uzaklaştırılarak, Bouin solüsyonunda +4°C de 36 saat süreyle tespit edildi. Bu işlemi müteakip doku örnekleri rutin histolojik prosedürlerden geçirilerek parafine gömüldü. Parafin bloklardan mikrotomda 5μm kalınlığında poly-L-lysin kaplı lamlara alınan doku kesitlerine, immunohistokimyasal boyama öncesinde, antijen maskelenmesi problemini gidermek için, mikrodalga ışınımlı "antijen retrieval" yöntemi uygulandı. İmmunohistokimyasal boyama Avidin-Biotin-Peroksidaz Kompleks (ABC) tekniği kullanılarak gerçekleştirildi. Bulgular: İncelenen tüm yaş grubu sıçanlarda; pozitif AR immunboyanma, epididimal hücrelerin sadece çekirdeklerinde gözlendi. AR pozitif hücrelerdeki boyanma yoğunluğu yaşa bağlı olarak değişim göstermedi. Kaput epididimiste pozitif AR immunboyanma; tubuler epitel hücreler (esas hücreler, bazal hücreler ve apikal hücreler) ve peritubuler düz kas hücrelerinde görüldü. Epitel hücrelerdeki boyanma yoğunluğu düz kas hücrelerine göre daha belirgindi. Epitel dokuda ise, esas hücrelerdeki boyanma yoğunluğu bazal ve apikal hücrelere göre daha güçlüydü. Sonuç: Epididimal hücrelerdeki AR yoğunluğunun, yaşa bağlı olarak değişmediği bulgusu; androjenin epididimisin postnatal farklılaşması ve epitel dokusunun yapısının korunabilmesi için gerekli olduğunu göstermektedir. Esas hücrelerin boyanma yoğunluğunun daha güçlü olması, bu hücrelerin androjen sitümülasyonuna daha duyarlı olduğu şeklinde yorumlanabilir.

Anahtar Kelimeler

• Androjen reseptör, immunohistokimya, kaput epididimis, antijen retrieval, sıçan

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