Article| Volume 46, ISSUE 1, P165-178, January 2023

The sirtuin 1 activator YK 3-237 stimulates capacitation-related events in human spermatozoa

  • David Martin-Hidalgo
    Corresponding author.
    Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Universidad de Extremadura, Avda de la Universidad s/n, Cáceres 10003, Spain

    Unit for Multidisciplinary Research in Biomedicine (UMIB), Department of Anatomy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal

    Unidad de Investigación, Complejo Hospitalario Universitario de Cáceres, Avenida Pablo Naranjo s/n, Cáceres 10003, Spain
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  • Lauro González-Fernández
    Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Universidad de Extremadura, Avda de la Universidad s/n, Cáceres 10003, Spain
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  • M. Julia Bragado
    Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Universidad de Extremadura, Avda de la Universidad s/n, Cáceres 10003, Spain
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  • Luis J. Garcia-Marin
    Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Universidad de Extremadura, Avda de la Universidad s/n, Cáceres 10003, Spain
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  • Marco G. Alves
    Unit for Multidisciplinary Research in Biomedicine (UMIB), Department of Anatomy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal
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  • Pedro F. Oliveira
    7QOPNA and LAQV, Department of Chemistry, University of Aveiro, Aveiro, Portugal
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      • SIRT1 is localized in the connecting piece of human spermatozoa.
      • Lysine acetylation is mainly found along the tail of human spermatozoa.
      • SIRT1 activation increases tyrosine phosphorylation.
      • SIRT1 activation improves human sperm responsiveness to Ca2+ ionophore challenge.


      Research question

      Does sirtuin-1 (SIRT1) have a role in the human spermatozoa capacitation process?


      Human spermatozoa were incubated for 6 h in a capacitating medium in presence or absence of the specific SIRT1 activator, YK 3-237. Several sperm parameters were determined by flow cytometry: viability, acrosome reaction and mitochondria membrane status. Sperm motility was determined objectively by computer-assisted semen analysis. Sperm capacitation status was evaluated by the extent of protein tyrosine phosphorylation and by the percentage of spermatozoa with the acrosome reacted by a calcium ionophore challenge.


      SIRT1 was detected in the connecting piece of human spermatozoa where a lysine acetylation pattern was mainly found along the sperm tail. SIRT1 activation accelerates the occurrence of a phenotype associated with human sperm capacitation, with no differences seen in the lysine acetylation pattern. After 1 h of co-incubation of YK 3-237 with human spermatozoa, tyrosine phosphorylation levels were comparable to control levels after 6 h of incubation in capacitating conditions. In addition, the activator improved sperm responsiveness to a Ca2+ ionophore (A23187) challenge determined by an increase in acrosome-reacted spermatozoa (P = 0.025). Importantly, sperm viability and mitochondrial activity-related parameters assessed by flow cytometry were not affected by YK 3-237.


      YK 3-237 induces capacitation-related events in human spermatozoa such an increase of tyrosine phosphorylation levels and acrosome-reacted spermatozoa after the ionophore challenge. Together, these results show that YK 3-237 affects human spermatozoa capacitation-related events by a mechanism independent of protein lysine acetylation but dependent on bicarbonate and calcium.


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      David Martín-Hidalgo is a biology reproductive researcher with a veterinary background. He is interested in improving the outcome of assisted reproductive technology in mammals, mainly humans and domestic animals. His research and expertise have been focused on the study of sperm metabolism and intracellular pathways as a tool to overcome male infertility.
      Key message
      This is the first study to show the role of sirtuins in the human sperm capacitation pathway. Human spermatozoa SIRT1 activated by YK 3-237 enhances capacitation-related events by bringing forward and increasing the levels of protein tyrosine phosphorylation and increasing the percentage of spermatozoa acrosome reacted after calcium ionophore challenge.