Immunohistochemical Localization of Ghrelin in Testicular Tissues of Holstein Bulls
الموضوعات :آ. بایزیدی آذر 1 , ب. شکرالهی 2
1 - Department of Animal Science, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran
2 - Department of Animal Science, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran
الکلمات المفتاحية: Ghrelin, Testis, Holstein bulls, leydig cell, sertoli cell,
ملخص المقالة :
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.
Barreiro M.L., Gaytan F., Caminos J.L., Pinilla L., Casanueva F.F., Aguilar E., Dieguez C. and Tena-Semper M. (2002).Cellular location and hormonal regulation of ghrelin expression in rat testis. Biol. Reprod. 67, 1768-1776.
Barreiro M.L., Suominen J.S., Gaytan F., Pinilla L., Chopin L.K., Casanueva F.F., Dieguez C., Aguilar E., Toppari J. and Tena-Sempere M. (2003). Developmental, stage-specific and hormonally regulated expression of growth hormone secretagogue receptor messenger RNA in rat testis. Biol. Reprod. 68, 1631-1640.
Barreiro M.L. and Tena-Sempere M. (2004). Ghrelin and reproduction: a novel signal linking energy status and fertility? Mol. Cell. Endocrinol. 226, 1-9.
Casanueva F.F. and Dieguez C. (1999). Neuroendocrine regulation and actions of leptin. Front. Neuroendocrinol. 20, 317-323.
Ciampani T., Fabbri A., Isidori A. and Dufau M.L. (1992). Growth hormone-releasing hormone is produced by rat leydig cell in culture and acts as a positive regulator of leydig cell function. Endocrinology. 131, 2785-2792.
Cowley M.A., Smith R.G., Diano S., Tschop M., Pronchuk N., Grove K.L., Strasburger C.J., Bidlingmaier M., Esterman M., Heiman M.L., Garcia-Segura L.M., Nillni E.A., Mendez P., Low M.J., Sotonyi P., Friedman J.M., Liu H., Pinto S., Colmers W.F., Cone R.D. and Horvath T.L. (2003). The distribution and mechanism of action of ghrelin in the CNS demonstrates a novel hypothalamic circuit regulating energy homeostasis. Neuron. 37, 649-661.
Dupont J., Maillard V., Coyral-Castel S., Rame C. and Froment P. (2010). Ghrelin in female and male reproduction. Int. J. Pept. 1, 1-8.
Fernandez-Fernandez R., Tena-Sempere M., Aguilar E. and Pinilla L. (2004). Ghrelin effects on gonadotropin secretion in male and female rats. Neurosci. Lett. 362, 103-107.
Furuta M., Funabashi T. and Kimura F. (2001). Intracerebroventricular administration of ghrelin rapidly suppresses pulsatile luteinizing hormone secretion in ovariectomized rats. Biochem. Biophys. Res. Commun. 288, 780-785.
Garcia M.C., Lopez M., Alvarez C.V., Casanueva F., Tena-Sempere M. and Dieguez C. (2007). Role of ghrelin in reproduction. Reproduction. 133, 531-540.
Gaytan F., Barreiro M.L., Chopin L.K., Herington A.C., Morales C., Pinilla L., Casanueva F.F., Aguilar E., Dieguez C. and Tena-Sempere M. (2003). Immunolocalization of ghrelin and its functional receptor, the type 1a growth hormone secretagogue receptor, in the cyclic human ovary. J. Clin. Endocrinol. Metab. 88, 879-887.
Ishikawa T., Fujioka H., Ishimura T., Takenaka A. and Fujisawa M. (2007). Ghrelin expression in human testis and serum testosterone level. J. Androl. 28, 320-324.
Kojima M., Hosoda H., Date Y., Nakazato M., Matsuo H. and Kangawa K. (1999). Ghrelin is a growth-hormone-releasing acylated peptide from stomach. Nature. 402, 656-560.
Lorenzi T., Meli R., Marzioni D., Morroni M., Baragli A., Castellucci M., Gualillo O. and Muccioli G. (2009). Ghrelin: a metabolic signal affecting the reproductive system. Cytokine. Growth. Factor. Rev. 20, 137-152.
Lukaszyk A., Kotwicka M., Jankowska A., Kasprzak A., Rucinski M., Sterzynska K., Ziolkowska A., Sawinski P. and Ruchala M. (2012). Expression of ghrelin receptor (GHSR-1a) in rat epididymal spermatozoa and the effects of its activation. Reprod. Biol. 12, 293-300.
Lukaszyk A., Kotwicka M., Jankowska A., Kasprzak A., Rucinski M., Sterzynska K., Ziolkowska A., Sawinski P. and Ruchala M. (2012). Expression of ghrelin receptor (GHSR-1a) in rat epididymal spermatozoa and the effects of its activation. Reprod. Biol. 12, 293-300.
Miller D.W., Harrison J.L., Brown Y.A., DoyleU., Lindsay A., Adam C.L. and Lea R.G. (2005). Immunohistochemical evidence for an endocrine/paracrine role for ghrelin in the reproductive tissues of sheep. Reprod. Biol. Endocrinol. 3, 60-71.
Muccioli G., Lorenzi T., Lorenzi M., Ghe C., Arnoletti E., Raso G.M., Castellucci M., Gualillo O. and Meli R. (2011). Beyond the metabolic role of ghrelin: a new player in the regulation of reproductive function. Peptides. 32, 2514-2521.
Tena-Sempere M., Barreiro M.L., Gonzalez L.C., Gaytan F., Zhang F.P., Caminos J.E., Pinilla L., Casanueva F.F., Dieguez C. and Aguilar E. (2002). Novel expression and functional role of ghrelin in rat testis. Endocrinology. 143, 717-725.
Wren A.M., Small C.J., Ward H.L., Murphy K.G., Dakin C.L., Taheri S., Kennedy A.R., Roberts G.H., Morgan D.G., Ghatei M.A. and Bloom S.R. (2000). The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion. Endocrinology. 141, 4325-4338.
Wenlong Z., Zhihai L., Juan S. and Shulin C. (2008). Expression of ghrelin in the porcine hypothalamo-pituitary-ovary axis during the oestrous cycle. Anim. Reprod. Sci. 109, 356-367.
