Immunohistochemical Localization of Ghrelin in Testicular Tissues of Holstein Bulls
محورهای موضوعی : Camel
آ. بایزیدی آذر
1
(
Department of Animal Science, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran
)
ب. شکرالهی
2
(
Department of Animal Science, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran
)
کلید واژه: Ghrelin, Testis, Holstein bulls, leydig cell, sertoli cell,
چکیده مقاله :
Ghrelin, a 28-amino acid acylated peptide, was identified as the endogenous ligand for the growth hormone (GH) secretagogue receptor and its role in reproduction has recently been established. It is believed that the site of ghrelin expression in testes may indicate its role in local regulation. This study investigated the immunohistochemical (IHC) localization of ghrelin in Holstein bull testis using a monoclonal anti-ghrelin antibody as primary antibody and donkey anti-rabbit IgG horseradish peroxidase (HRP) polyclonal antibody as secondary antibody. Samples of testis were collected from three Holstein bulls aged of 1.5 to 2 years old and preserved in 10% formalin for posterior inclusion in paraffin. Histological sections with 5 micron in thickness were prepared for IHC. Immunoreactions were assessed for spermatogenic cells and leydig and sertoli cells. It is believed that the site of ghrelin expression in the spermatogenesis process, leydig and sertoli cells, may indicate its role in local regulations. This is one of the first studies to provide molecular evidence for the presence of ghrelin within testicular tissue cells of Holstein bulls.
گرلین، پپتید 28 اسید آمینه آسیله شده، به عنوان لیگاند اندوژنوسی برای گیرنده ترشحی هورمون رشد (GH) و نقش آن در تولید مثل که اخیراً منتشر شده شناسایی شده است. عقیده بر این است که محل بیان گرلین در بیضه‌ها ممکن است نقش آن را در تنظیم موضعی نشان دهد. در این مطالعه موقعیت‌یابی ایمنوهیستوشیمی (IHC) گرلین در بافت بیضه گاو هلشتاین با استفاده از آنتی‌بادی مونوکلونال ضد گرلین به عنوان آنتی‌بادی اولیه و آنتی‌بادی پلی‌کلونال دانکی ضد IgG خرگوشی گونژوگه شده با هورس رادیش پراکسیداز (HRP) به عنوان آنتی‌بادی ثانویه انجام گرفت. نمونه‌های بیضه از سه گاو هلشتاین 2-5/1 ساله جمع‌آوری شدند و جهت قرار گرفتن در پارافین در مرحله بعدی در فرمالین 10 درصد نگهداری شدند. مقاطع بافتی با ضخامت 5 میکرون برای آزمایش IHC تهیه شد. واکنش‌های ایمنی برای سلول‌های اسپرماتوژنیک و سلول‌های لایدیگ و سرتولی ارزیابی شدند. عقیده بر این است که محل بیان گرلین در مراحل اسپرماتوژنز، سلول‌های لایدیگ و سرتولی ممکن است نقش آن را در تنظیم موضعی نشان دهد. این یکی از اولین تحقیقاتی است که شواهد مولکولی را برای وجود گرلین در سلول‌های بافت بیضه گاو هلشتاین فراهم می‌کند.
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