بیان ژن آدیپونکتین و گیرنده 2 آن (AdipoR2) در مجرای تولیدمثلی بره‌های نر افشاری پیش از بلوغ

نوع مقاله : علمی پژوهشی- فیزیولوژی

نویسندگان

1 گروه علوم دامی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران

2 گروه علوم دامی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران.

چکیده

آدیپونکتین هورمونی است که عمدتا از بافت چربی ترشح می‌شود و شواهد اخیر حاکی از نقش آن در تنظیم سیستم تولیدمثلی پستانداران است. هدف از این پژوهش بررسی پروفیل بیان ژن آدیپونکتین و گیرنده 2 آن (AdipoR2) در نواحی مختلف مجرای تولیدمثلی بره‌های نر نژاد افشاری در دوره‌ پیش از بلوغ بود. بدین منظور، 20 رأس بره نر نابالغ نژاد افشاری به چهار گروه سنی مختلف شامل؛ گروه‌های سنی 1 تا 2، 2 تا 3، 3 تا 4 و 4 تا 5 ماه اختصاص یافتند، سپس دام‌ها به روش جراحی اخته شدند و از چهار بافت سر، بدنه، دم اپیدیدیم و پارانشیم بیضه هر دام نمونه‌برداری به عمل آمد. بیان دو ژن آدیپونکتین و AdipoR2 با استفاده از تکنیک Real Time-QPCR در هر نمونه بافت بررسی شد. بطور خلاصه، بیان مداوم ژن‌های آدیپونکتین و AdipoR2 در تمام مراحل رشد، قبل و هنگام بلوغ مشاهده شد. اگرچه میزان بیان آدیپونکتین و AdipoR2 در بین گروه‌های سنی مختلف به لحاظ آماری متفاوت بود، با این حال تفاوت آماری معنی‌داری در مقدار بیان آن‌ها در بین چهار بافت مختلف پارانشیم بیضه و سر، بدنه و دم اپیدیدیم بره‌های نر نابالغ مشاهده نشد. نتایج بررسی حاضر نشان داد که آدیپونکتین و گیرنده 2 آن (AdipoR2) به طور مداوم در مجرای تولیدمثلی بره‌های نر نابالغ بیان می‌شوند و سطح بیان آنها با افزایش سن تغییر می‌کند. با توجه به تازگی این موضوع، نتایج این مطالعه می‌تواند زمینه ساز مطالعات بیشتر در این زمینه مهم باشد.
واژه های کلیدی: قوچ افشاری، بیضه، تولیدمثل، آدیپونکتین

کلیدواژه‌ها


عنوان مقاله [English]

Gene Expression of Adiponectin and Its Receptor2 (AdipoR2) in Reproductive Tract of Growing Prepubertal Afshari Male Lambs

نویسندگان [English]

  • Mohammad Dorounaki 1
  • Behnam Rostami 1
  • Mohammad Taher Harkinezhad 1
  • Mohammad Hossein Shahir 2
1 Department of Animal Science, Zanjan Faculty of Agriculture, Zanjan, Iran
2 Department of Animal Sciences, Faculty of Agriculture, Zanjan University, Zanjan, Iran.
چکیده [English]

Introduction Adiponectin, a novel adipocytokine mainly produced from adipocytes, acts through binding to its two functional receptors, AdipoR1 and AdipoR2. Some evidence has already suggested that adiponectin can be involved in the regulation of male reproductive functions. Our purpose was to detect of adiponectin and AdipoR2 mRNA transcripts in the reproductive tissues of growing pre-pubertal male lambs as well as adult rams.
Materials and Methods At first, Afshari prepubertal male lambs (n=20), with no apparent abnormalities, were allocated to four different age groups (n=5/age group) including; 1-2 (Group 1), 2-3 (Group 2), 3-4 (Group 3), and 4-5 (Group 4) months of age, afterward were surgically castrated. At a local slaughterhouse, reproductive tissues from five adult Afshari rams (3-4 years-old), with no evident abnormalities, were also collected within 20 minutes after the rams were slaughtered. Both fresh whole testes for lambs and those for adult rams were instantly transported to the laboratory on ice and were frozen in the liquid nitrogen and stored at −80°C until analysis. Total RNA extracted from each tissue sample, converted to cDNA and finally, the relative expression levels of adiponectin and AdipoR2 transcripts were assessed by Quantitative Real time-PCR. Comparative ΔΔCt method was used for quantification of Real-Time PCR outputs. By GenEx enterprise software, obtained fold changes were statistically analyzed.
Results and Discussion By real-time PCR, local gene expression of adiponectin and AdipoR2 was demonstrated in testis parenchyma and three different parts of epididymis (caput, corpus and cauda) for both immature male lambs and adult rams. The results showed that adiponectin and AdipoR2 levels persistently changed during pre-pubertal and pubertal developmental stages. There were no significant differences in adiponectin, and AdipoR2 mRNA levels among various tissues (p>0.05); but, mRNA quantities significantly varied among different age groups (p

کلیدواژه‌ها [English]

  • Adiponectin
  • Afshari Ram
  • Gene expression
  • reproduction
  • Testis
1- Bauche, I. B., E. l. Ait., S. Mkadem., R. Rezsohazy., T. Funahashi., N. Maeda., L. M. Miranda, and S. M. Brichard. 2006. Adiponectin downregulates its own production and the expression of its AdipoR2 receptor in transgenic mice. Biochemical and Biophysical Research Communications, 345: 1414-1424.
2- Bjursell, M., A. Ahnmark., Y. M. Bohlooly., L. William-Olsson., M. Rhedin., X. R. Peng., K. Ploj., A. K. Gerdin., G. Arnerup., A. Elmgren., A. L. Berg., J. Oscarsson, and D. Linden. 2007. Opposing effects of adiponectin receptors 1 and 2 on energy metabolism. Diabetes, 56: 583-593.
3- Caminos, J. E., R. Nogueiras., F. Gaytan., R. Pineda., C. R. Gonzalez., M. L. Barreiro., J. P. Castano., M. M. Malagon., L. Pinilla., J. Toppari., C. Dieguez, and M. Tena-Sempere. 2008. Novel Expression and direct effects of adiponectin in the rat testis. Endocrinology, 149:3390-3402.
4- Ceddia, R. B., R. Somwar., A. Maida., X. Fang., G. Bikopoulos, and G. Sweeney. 2005. Globular adiponectin increases GLUT4 translocation and glucose uptake but reduces glycogen synthesis in rat skeletal muscle cells. Diabetologia, 48: 132-139.
5- Chabrolle, C., L. Tosca., S. Crochet., S. Tesseraud, and J. Dupont. 2007. Expression of AdipoR1 and AdipoR2 in chicken ovary: potential role in ovarian steroidogenesis. Domestic Animal Endocrinology, 33: 480-487.
6- Chabrolle, C., L. Tosca, and J. Dupont. 2007. Regulation of adiponectin and its receptors in rat ovary by human chorionic gonadotrophin treatment and potential involvement of adiponectin in granulosa cell steroidogenesis. Reproduction, 133: 719-731.
7- Chappaz, E., M. S. Albornoz., D. Campos., L. Chea., M. F. Palin, and D. Bruce. 2008. Adiponectin enhances in vitro development of swine embryos. Domestic Animal Endocrinology, 35: 198-207.
8- Combs, T. P., A. H. Berg., M. W. Rajala., S. Klebanov., P. Iyengar., J. C. Jimenez Chillaron., M. E. Patti., S. L. Klein., R. S. Weinstein, and P. E. Scherer. 2003. Sexual differentiation, pregnancy, calorie restriction, and aging affect the adipocyte-specific secretory protein adiponectin. Diabetes, 52: 268-276.
9- Dai, M. H., T. Xia., G. D. Zhang., X. D. Chen., L. Gan, and S. Q. Feng. 2006. Cloning, expression and chromosome localization of porcine adiponectin and adiponectin receptors genes. Domestic Animal Endocrinology, 30: 117-125.
10- Dupont, J., C. Chabrolle., C. Rame., L. Tosca, and S. Coyral-Castel. 2008. Role of the peroxisome proliferator-activated receptors, adenosine monophosphate-activated kinase, and adiponectin in the ovary. PPAR Research, 2008:1-9.
11- Kadowaki, T. and T. Yamauchi. 2005. Adiponectin and adiponectin receptors. Endocrine Reviews, 26: 439-51.
12- Kershaw, E. E. and J. S. Flier. 2004. Adipose tissue as an endocrine organ. Journal of Clinical Endocrinology and Metabolism, 89: 2548-2556.
13- Lagaly, D. V., P. Y. Aad., J. A. Grado-Ahuir., L. B. Hulsey, and L. J. Spicer. 2008. Role of adiponectin in regulating ovarian theca and granulosa cell function. Molecular and Cellular Endocrinology, 284: 38-45.
14- Ledoux, S., D. B. Campos., F. L. Lopes., M. Dobias-Goff., M. F. Palin, and B. D. Murphy. 2006. Adiponectin induces periovulatory changes in ovarian follicular cells. Endocrinology, 147: 5178-5186.
15- Lord, E., S. Ledoux., B. D. Murphy., D. Beaudry, and M. F. Palin. 2005. Expression of adiponectin and its receptors in swine. Journal of Animal Science, 83: 565-578.
16- Ma, H., V. Gomez., L. Lu., X. Yang., X. Wu, and S. Y. Xiao. 2009. Expression of adiponectin and its receptors in livers of morbidly obese patients with non-alcoholic fatty liver disease. Journal of Gastroenterology and Hepatology, 24(2): 233-237.
17- Maddineni, S., S. Metzger., O. Ocon., G. I. I. I. Hendricks, and R. Ramachandran. 2005. Adiponectin gene is expressed in multiple tissues in the chicken: food deprivation influences adiponectin messenger ribonucleic acid expression. Endocrinology, 1464: 250-256.
18- Maeda, N., I. Shimomura., K. Kishida., H. Nishizawa., M. Matsuda., H. Nagaretani., N. Furuyama., H. Kondo., M. Takahashi., Y. Arita., R. Komuro., N. Ouchi., S. Kihara., Y. Tochino., K. Okutomi., M. Horie., S. Takeda., T. Aoyama., T. Funahashi, and Y. Matsuzawa. 2002. Diet-induced insulin resistance in mice lacking adiponectin/ACRP30. Nature Medicine, 8: 731-737.
19- Maillard, V., S. Uzbekova., F. Guignot., C. Perreau., C. Rame., S. Coyral-Castel, and J. Dupont. 2010. Effect of adiponectin on bovine granulosa cell steroidogenesis, oocyte maturation and embryo development. Reproductive Biology and Endocrinology, 8:23-38.
20- Ocoǐn-Grove, O. M., S. M. Krzysik-Walker., S. R. Maddineni., G. L. Hendricks, and R. Ramachandran. 2008. Adiponectin and its receptors are expressed in the chicken testis: influence of sexual maturation on testicular ADIPOR1 and ADIPOR2 mRNA abundance. Reproduction, 136: 627-638.
21- Paschke, L., T. Zemleduch., M. Rucinski., A. Ziolkowska., M. Szyszka, and L. K. Malendowicz. 2010. Adiponectin and adiponectin receptor system in the rat adrenal gland: ontogenetic and physiologic regulation, and its involvement in regulating adrenocortical growth and steroidogenesis. Peptides, 31(9): 1715-1724.
22- Pfaffl, M. W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research, 29: e45.
23- Rahmanifar, F, and M. R. Tabandeh. 2012. Adiponectin and its receptors gene expression in the reproductive tract of ram. Small Ruminant Research, 105: 263-267.
24- Ramachandran, R., O. M. Ocon-Grove, and S. L. Metzger. 2007. Molecular cloning and tissue expression of chicken AdipoR1 and AdipoR2 complementary deoxyribonucleic acids. Domestic Animal Endocrinology, 33 (1): 19-31.
25- Rodriguez-Pacheco, F., A. J. Martinez-Fuentes., S. Tovar., L. Pinilla., M. Tena-Sempere., C. Dieguez., J. P. Castano, and M. M. Malagon. 2007. Regulation of pituitary cell function by adiponectin. Endocrinology, 148: 401-410.
26- Schaffler, A., J. Scholmerich, and B. Salzberger. 2007. Adipose tissue as an immunological organ: Toll-like receptors, C1q/TNFs and CTRPs. Trends in Immunology, 28: 393-399.
27- Tabandeh, M. R., N. Golestani., M. Kafi., A. Hosseini., M. Saeb, and P. Sarkoohi. 2012. Gene expression pattern of adiponectin and adiponectin receptors in dominant and atretic follicles and oocytes screened based on brilliant cresyl blue staining. Animal Reproduction Science, 131: 30-40.
28- Wena, J. P., W. S. Lv., J. Yang., A. F. Nie., X. B. Cheng, and Y. Yang. 2008. Globular adiponectin inhibits GnRH secretion from GT1-7 hypothalamic GnRH neurons by induction of hyperpolarization of membrane potential. Biochemical and Biophysical Research Communications, 371: 756-761.
29- Yamauchi, T., J. Kamon., Y. Ito., A. Tsuchida., T. Yokomizo., S. Kita., T. Sugiyama., M. Miyagishi., K. Hara., M. Tsunoda., K. Murakami., T. Ohteki., S. Uchida., S. Takekawa., H. Waki., N. H. Tsuno., Y. Shibata., Y. Terauchi., P. Froguel., K. Tobe., S. Koyasu., K. Taira., T. Kitamura., T. Shimizu., R. Nagai, and T. Kadowaki. 2003. Cloning of adiponectin receptors that mediate antidiabetic metabolic effects. Nature, 423(6941): 762-769.
CAPTCHA Image