Evidence of melatonin synthesis in the ram reproductive tract

Evidence of melatonin synthesis in the ram reproductive tract

Summary Melatonin is a ubiquitous molecule found in a wide range of fluids, one of them being ram seminal plasma, in which it can reach higher concentrations than those found in blood, suggesting an extrapineal secretion by the reproductive tract. In order to identify the source of the melatonin fou...

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Bibliographic Details
Published in:Andrology (Oxford) Vol. 4; no. 1; pp. 163 - 171
Main Authors: Gonzalez‐Arto, M., Hamilton, T. R dos S., Gallego, M., Gaspar‐Torrubia, E., Aguilar, D., Serrano‐Blesa, E., Abecia, J.A., Pérez‐Pé, R., Muiño‐Blanco, T., Cebrián‐Pérez, J. A., Casao, A.
Format: Journal Article
Language:English
Published: England Wiley Subscription Services, Inc 01-01-2016
Subjects:
AANAT
Acetylserotonin O-Methyltransferase - metabolism
Animals
Arylalkylamine N-Acetyltransferase - metabolism
ASTM
Epididymis - metabolism
extrapineal
Leydig Cells - metabolism
Male
Melatonin - biosynthesis
Melatonin - metabolism
Semen - metabolism
seminal plasma
Sheep
Spermatids - metabolism
Spermatocytes - metabolism
testes
Testis - cytology
Testis - metabolism
seminal plasma
AANAT
ASTM
extrapineal
testes
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Summary:Summary Melatonin is a ubiquitous molecule found in a wide range of fluids, one of them being ram seminal plasma, in which it can reach higher concentrations than those found in blood, suggesting an extrapineal secretion by the reproductive tract. In order to identify the source of the melatonin found in ram seminal plasma, we first tried to determine whether the melatonin levels were maintained during the day. For this purpose, melatonin concentrations were measured in seminal plasma obtained from first ejaculates of six rams at 6:00 a.m. in total darkness, at 10:00 a.m. and at 14:00 p.m. The melatonin concentration was higher (p < 0.05) in ejaculates collected at 6:00 a.m. than at 10:00 and 14:00. There was no statistical difference between the latter. To further corroborate an extrapineal secretion of melatonin, the presence of the two key enzymes involved in melatonin synthesis, arylalkylamine‐N‐acetyltransferase (AANAT) and N‐acetylserotonin‐O‐methyltransferase (ASMT) was analyzed by RT‐PCR, q‐PCR and Western‐blot in ram testes, epididymis, and accessory glands. The RT‐PCR showed the presence of the m‐RNA codifying both AANAT and ASTM in all the tissues under study, but the q‐PCR and Western‐blot revealed that gene expression of these enzymes was significantly higher in the testis (p < 0.05). Immunohistochemistry confirmed the presence of AANAT and ASMT in the testis and revealed that they were found in the Leydig cells, spermatocytes, and spermatids. Also, measurable levels of melatonin were found in testicular tissue and the tail of the epididymis. In conclusion, our study indicates that the testes are one of the likely sources of the high levels of melatonin found in ram seminal plasma, at least during the day.
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ISSN:2047-2919
2047-2927
DOI:10.1111/andr.12117