BACKGROUND: A recent study of several compounds, structurally related to amphetamine, provided evidence that mammalian spermatozoa might have adrenergic receptors able to regulate cAMP production. The present study investigated this possibility using physiological and immunochemical analyses of mouse and human spermatozoa. METHODS: Antibodies specific for different receptor subtypes were used for Western blotting of mouse and human sperm lysates and for immunocytochemical evaluation of whole mouse and human spermatozoa. Uncapacitated and capacitated mouse spermatozoa were incubated with specific agonists and antagonists for alpha2-, beta1-, beta2- and beta3-adrenergic receptors for approximately 35 min and then assessed using chlortetracycline (CTC) fluorescence. RESULTS: Western blotting revealed proteins of the correct size for all these receptors; immunolocalization indicated their presence on the head, especially acrosomal and neck regions, and flagellum of both mouse and human spermatozoa. CTC results indicated significant responses to agonists for all of the beta-receptors in uncapacitated cells, with agonist effectiveness being beta1 > beta2 > beta3; relevant antagonists blocked responses. In contrast, an agonist and antagonist for alpha2-receptors acted only on capacitated spermatozoa. CONCLUSION: These experiments provide the first good evidence that mammalian spermatozoa have both beta-adrenergic receptors, known to stimulate cAMP production by membrane-associated adenylyl cyclases (mACs), and alpha2-adrenergic receptors, known to inhibit cAMP production by mACs. Responses are capacitation state dependent and provide a mechanism for inhibiting spontaneous acrosome reactions and helping to maintain fertilizing ability. These results suggest that the use of amphetamine-related compounds, either for medical or for social reasons, might have an unexpected positive impact on fertility.
BACKGROUND: A recent study of several compounds, structurally related to amphetamine, provided evidence that mammalian spermatozoa might have adrenergic receptors able to regulate cAMP production. The present study investigated this possibility using physiological and immunochemical analyses of mouse and human spermatozoa. METHODS: Antibodies specific for different receptor subtypes were used for Western blotting of mouse and human sperm lysates and for immunocytochemical evaluation of whole mouse and human spermatozoa. Uncapacitated and capacitated mouse spermatozoa were incubated with specific agonists and antagonists for alpha2-, beta1-, beta2- and beta3-adrenergic receptors for approximately 35 min and then assessed using chlortetracycline (CTC) fluorescence. RESULTS: Western blotting revealed proteins of the correct size for all these receptors; immunolocalization indicated their presence on the head, especially acrosomal and neck regions, and flagellum of both mouse and human spermatozoa. CTC results indicated significant responses to agonists for all of the beta-receptors in uncapacitated cells, with agonist effectiveness being beta1 > beta2 > beta3; relevant antagonists blocked responses. In contrast, an agonist and antagonist for alpha2-receptors acted only on capacitated spermatozoa. CONCLUSION: These experiments provide the first good evidence that mammalian spermatozoa have both beta-adrenergic receptors, known to stimulate cAMP production by membrane-associated adenylyl cyclases (mACs), and alpha2-adrenergic receptors, known to inhibit cAMP production by mACs. Responses are capacitation state dependent and provide a mechanism for inhibiting spontaneous acrosome reactions and helping to maintain fertilizing ability. These results suggest that the use of amphetamine-related compounds, either for medical or for social reasons, might have an unexpected positive impact on fertility.
Authors: Javier A Urra; Franz Villaroel-Espíndola; Alejandra A Covarrubias; Joan Enric Rodríguez-Gil; Alfredo Ramírez-Reveco; Ilona I Concha Journal: PLoS One Date: 2014-11-17 Impact factor: 3.240