Melatonin reduces cAMP-stimulated capacitation of ram spermatozoa
Silvia Gimeno-Martos
A , Adriana Casao A , Marc Yeste B , José A. Cebrián-Pérez A , Teresa Muiño-Blanco A and Rosaura Pérez-Pé A C
+ Author Affiliations
- Author Affiliations
A Department of Biochemistry and Molecular and Cell Biology, Institute of Environmental Sciences of Aragón, School of Veterinary Medicine, University of Zaragoza, C/Miguel Servet 177, 50013, Zaragoza, Spain.
B Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, C/Maria Aurèlia Campany 69, Campus Montilivi, E-17003 Girona, Spain.
C Corresponding author. Email: rosperez@unizar.es
Reproduction, Fertility and Development 31(2) 420-431 https://doi.org/10.1071/RD18087
Submitted: 6 March 2018 Accepted: 7 August 2018 Published: 13 September 2018
Abstract
The presence of melatonin receptors on the surface of ram spermatozoa has led to speculation about melatonin having a role in sperm functionality. The aim of this study was to elucidate the mechanism through which melatonin regulates ram sperm capacitation induced by a cocktail containing cAMP-elevating agents. Cocktail samples capacitated in the presence of 1 µM melatonin showed lower percentages of capacitated spermatozoa (chlortetracycline staining; P < 0.001) together with a decrease in protein tyrosine phosphorylation (P < 0.01) and lower levels of reactive oxygen species (ROS) and cAMP (P < 0.05) compared with cocktail samples without the hormone. Determination of kinematic parameters, together with principal component and cluster analyses, allowed us to define four sperm subpopulations (SP). After 3 h of incubation with cAMP-elevating agents, the percentages of spermatozoa belonging to SP1 (high straightness) and SP4 (less-vigorous spermatozoa with non-linear motility) increased while SP2 and SP3 (rapid spermatozoa starting hyperactivation or already hyperactivated) decreased compared with the control sample. The presence of melatonin at 100 pM and 10 nM restored these subpopulations to values closer to those found in the control sample. These results indicate that melatonin at micromolar concentrations modulates ram sperm capacitation induced by cAMP-elevating agents, reducing ROS and cAMP levels, whereas at lower concentrations melatonin modifies motile sperm subpopulations. These findings warrant further studies on the potential use of melatonin for controlling capacitation in artificial insemination procedures.
Additional keywords: kinematic parameters, protein tyrosine phosphorylation, reactive oxygen species.
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