Induction of acrosome reaction by 4-Br-A23187 alters the glycoproteomic profile of boar spermatozoa.

Autor: Martín-Hidalgo D; Departamento de Fisiología, Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain., Izquierdo M; Centro de Investigaciones Científicas y Tecnológicas de Extremadura (CICYTEX), Badajoz, Spain., Garrido N; MED-Mediterranean Institute for Agriculture, Environment and Development & CHANGE-Global Change and Sustainability Institute, Escola Superior Agrária de Elvas, Departamento de Ciência Agrárias e Veterinárias, Elvas, Portugal., Bartolomé-García P; Centro de Selección y Reproducción Animal (CENSYRA), Badajoz, Spain., Macías-García B; Departamento de Medicina Animal, Grupo de Investigación Medicina Interna Veterinaria (MINVET), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain., González-Fernández L; Departamento de Bioquímica y Biología Molecular y Genética, Grupo de Investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Instituto de Investigación INBIO G+C, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain. Electronic address: lgonfer@unex.es.
Jazyk: angličtina
Zdroj: Theriogenology [Theriogenology] 2024 Jul 15; Vol. 223, pp. 108-114. Date of Electronic Publication: 2024 Apr 17.
DOI: 10.1016/j.theriogenology.2024.04.006
Abstrakt: Protein glycosylation is a post-translational modification involved in wide range of biological processes. In mammalian spermatozoa this modification has been identified in numerous proteins, and membrane glycoproteins are involved in the fertilization process. The objective of the present study was to identify changes in protein glycosylation after acrosome reaction (AR) induction using the 4-Br-A23187 ionophore. Our results showed that treatment with 10 μM of 4-Br-A23187 for 20 min significantly increased the percentage of live acrosome-reacted spermatozoa compared to the control (69.8 ± 0.8 vs. 6.4 ± 0.5; mean % ± SEM, respectively). Also, we observed an increase in 32 kDa tyrosine-phosphorylated protein (p32) and a decrease in serine/threonine phosphorylation of the protein kinase A substrates (phospho-PKA-substrates) after ionophore treatment. Furthermore, changes in glycosylated proteins following AR induction were analyzed using different HRP-conjugated lectins (GNA, DSA, and SNA), revealing changes in mannose and sialic acid residues. Proteomic analysis of isolated proteins using GNA lectin revealed that 50 proteins exhibited significantly different abundance (q-value < 0.01). Subsequent analysis using Uniprot database identified 39 downregulated and 11 upregulated proteins in the presence of 4-Br-A23187. Notably, six of these proteins were classified as transmembrane proteins, namely LRRC37A/B like protein 1 C-terminal domain-containing protein, Membrane metalloendopeptidase like 1, VWFA domain-containing protein, Syndecan, Membrane spanning 4-domains A14 and Serine protease 54. This study shows a novel protocol to induce acrosome reaction in boar spermatozoa and identifies new transmembrane proteins containing mannose residues. Further work is needed to elucidate the role of these proteins in sperm-oocyte fusion.
Competing Interests: Declaration of competing interest The authors have declared that no competing interests exist.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: