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• W2023017922 abstract "ObjectiveTo verify the presence of protein precursor pro-enkephalin (PENK) and met-enkephalin in human spermatozoa and to characterize the effects of exogenous and endogenous enkephalins on sperm motility.DesignWe carried out expression assays for met-enkephalin and its protein precursor PENK by reverse transcription–polymerase chain reaction (RT-PCR), Western blot, and immunofluorescence techniques in sperm cells and motility analysis after incubation of semen samples with met-enkephlin enzyme inhibitors and the opioid receptor antagonist naloxone. Met-enkephalin secretion was analyzed by flow cytometry.SettingAssisted reproduction unit and academic research laboratory.Patient(s)Semen from 50 normozoospermic healthy human donors.Intervention(s)Spermatozoa isolated from semen on discontinuous Percoll gradient (40%–80%) followed by a swim-up was used for all techniques.Main Outcome Measure(s)Immunoblotting blots, indirect immunofluorescence antibody assays, RT-PCR blots, flow cytometry plots, and percentage of motile sperm.Result(s)We found by RT-PCR and immunofluorescence that met-enkephalin and its protein precursor PENK are present in the head of human sperm cells. Endogenous met-enkephalin increased sperm motility, whereas the addition of exogenous met-enkephalin had a biphasic effect on motility, likely due to the activation of distinct receptor subtypes.Conclusion(s)We provide evidence for a new role of met-enkephalin as an endogenous mediator of sperm motility. This autocrine regulation of sperm function by the opioid system represents a new mechanism of regulation of male factor fertility and could be useful as an emerging target for male contraception. To verify the presence of protein precursor pro-enkephalin (PENK) and met-enkephalin in human spermatozoa and to characterize the effects of exogenous and endogenous enkephalins on sperm motility. We carried out expression assays for met-enkephalin and its protein precursor PENK by reverse transcription–polymerase chain reaction (RT-PCR), Western blot, and immunofluorescence techniques in sperm cells and motility analysis after incubation of semen samples with met-enkephlin enzyme inhibitors and the opioid receptor antagonist naloxone. Met-enkephalin secretion was analyzed by flow cytometry. Assisted reproduction unit and academic research laboratory. Semen from 50 normozoospermic healthy human donors. Spermatozoa isolated from semen on discontinuous Percoll gradient (40%–80%) followed by a swim-up was used for all techniques. Immunoblotting blots, indirect immunofluorescence antibody assays, RT-PCR blots, flow cytometry plots, and percentage of motile sperm. We found by RT-PCR and immunofluorescence that met-enkephalin and its protein precursor PENK are present in the head of human sperm cells. Endogenous met-enkephalin increased sperm motility, whereas the addition of exogenous met-enkephalin had a biphasic effect on motility, likely due to the activation of distinct receptor subtypes. We provide evidence for a new role of met-enkephalin as an endogenous mediator of sperm motility. This autocrine regulation of sperm function by the opioid system represents a new mechanism of regulation of male factor fertility and could be useful as an emerging target for male contraception." @default.
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• W2023017922 date "2012-09-01" @default.
• W2023017922 modified "2023-10-09" @default.
• W2023017922 title "Autocrine regulation of human sperm motility by the met-enkephalin opioid peptide" @default.
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• W2023017922 doi "https://doi.org/10.1016/j.fertnstert.2012.05.036" @default.
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