ProAKAP4 as Indicator of Long-Lasting Motility Marker in Post-Thaw Conditions in Stallions

Author:

Dordas-Perpinyà Marta1234ORCID,Yánez-Ortiz Iván25ORCID,Sergeant Nicolas67,Mevel Vincent1,Catalán Jaime2ORCID,Bruyas Jean-François1,Miró Jordi2ORCID,Briand-Amirat Lamia1ORCID

Affiliation:

1. Oniris, Nantes Veterinary College, Cedex 03, 44307 Nantes, France

2. Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Cerdanyola del Vallès, Spain

3. UVSQ, INRAE, BREED, Université Paris-Saclay, 78352 Jouy-en-Josas, France

4. Ecole Nationale Vétérinaire d’Alfort, BREED, 94700 Maisons-Alfort, France

5. School of Veterinary Medicine, Faculty of Medicine, Health and Life Sciences, International University of Ecuador, Quito 170901, Ecuador

6. INSERM, UMRS, University of Lille, 59000 Lille, France

7. SPQI, 4bioDx—Breeding Section, 59000 Lille, France

Abstract

ProAKAP4, a precursor of AKAP4 (A-kinase anchor protein) found in the flagellum of mammalian and non-mammalian spermatozoa, serves as a structural protein with established correlations to motility parameters across diverse species. This study aimed to determine the proAKAP4 level evolution in thawed stallion semen over a 3 h period, examining its correlation with motility descriptors and mitochondrial membrane potential. Utilizing sixteen ejaculates from four French warmblood stallions, this study involved maintaining thawed samples at 37 °C for 3 h, conducting proAKAP4 enzyme-linked immunosorbent assays (ELISA), computer-assisted sperm analysis (CASA), and mitochondrial membrane potential by JC-1 probe and flow cytometry at 0, 1, and 3 h post-thawing. The findings indicate significant positive correlations (p ≤ 0.05) between proAKAP4 levels and sperm total or progressive motility at all time points analyzed. Spermatozoa velocity descriptors (VAP, VCL, VSL) and spermatozoa lateral head displacement (ALH) display positive correlations (p ≤ 0.05) with ProAKAP4 at the 0 h post-thawing. ProAKAP4 concentration exhibits no discernible difference between batches with or without a cryoprotectant. Notably, proAKAP4 consumption remains insignificant within the initial hour after thawing but becomes significant (p ≤ 0.05) between 1 and 3 h post-thawing. In summary, proAKAP4 demonstrates positive correlations with total and progressive motility in stallion semen for up to 3 h after thawing, albeit showing a noticeable decrease starting from the first hour post-thawing, indicating a progressive consumption as a result of spermatozoa motile activity.

Funder

IFCE

Publisher

MDPI AG

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