Expression of ghrelin receptor (GHSR-1a) in rat epididymal spermatozoa and the effects of its activation |
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| Authors: | Andrzej Łukaszyk Małgorzata Kotwicka Anna Jankowska Aldona Kasprzak Marcin Ruciński Karolina Sterzyńska Agnieszka Ziółkowska Piotr Sawiński Marek Ruchala |
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| Affiliation: | 1. Department of Histology and Embryology, Poznań University of Medical Sciences, Poznań, Poland;2. Department of Cell Biology, Poznań University of Medical Sciences, Poznań, Poland;3. Department of Endocrinology, Metabolism and Internal Medicine, Poznań University of Medical Sciences, Poznań, Poland;2. Department of Endocrinology, Institute of Zoology, Jagiellonian University;3. Department of Experimental Hematology, Institute of Zoology, Jagiellonian University, Krakow, Poland;2. Morphological Sciences Department, Faculty of Veterinary Sciences, National University of Litoral (FCV-UNL), Esperanza, Santa Fe, Argentina;3. Argentine National Research Council (CONICET);1. Department of Biomedical Sciences, Applied Biology Section, Italy;2. Interdepartmental Centre for Research and Therapy of Male Infertility, University of Siena, Siena, Italy;3. Rinaldi Fontani Institute, Firenze, Italy;4. Department of Pathology and Veterinary Clinic, Surgery Clinic Section, University of Sassari, Sassari, Italy;5. AGRIS, Department of Research for Increased Stallion Reproduction, Ozieri, Sassari, Italy |
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| Abstract: | In this study we demonstrated the expression of the ghrelin receptor GHSR-1a in rat spermatids and epididymal spermatozoa, as well as some effects of ghrelin on the spermatozoa in vitro. For the demonstration of GHSR-1a the immunocytochemical, immunofluorescence and Western blotting techniques were applied using three different types of antibodies. The response of spermatozoa to ghrelin was tested in a series of in vitro experiments and their effects were evaluated using confocal microscopy and flow cytometry. GHSR-1a protein was found as expressed in the Golgi and acrosomes of spermatids and acrosome regions or the head cell membrane of epididymal spermatozoa. The GHSR-1a expression in spermatozoa was also confirmed by Western blot. No differences were found in percentage of spermatozoa showing annexin-V binding and expression of active form caspase-3 between control and ghrelin-treated spermatozoa. This result may indicate no pro-apoptotic effects of ghrelin neither at 10?9 nor 10?6 mol/L concentration. Ghrelin (10?6 mol/L) increased free intracellular calcium ion concentration in the rat spermatozoa. Moreover, stimulation with 10?6 mol/L ghrelin increased, while 10?4 mol/L ghrelin decreased the number of spermatozoa showing progressive motility. In conclusion, the expression of the GHSR-1a receptor in spermatozoa, as well as ghrelin influences on sperm motility and intracellular calcium ion concentration suggest that such biological effects of ghrelin may be produced under in vivo conditions. |
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