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Updated: Jul 12, 2025
Phosphopeptide Analysis of Rodent Epididymal Spermatozoa
Published on: December 30, 2014
Processing of boar spermatozoa with phosphate-buffered saline at 4˚C induces an increase in 32 kDa
David Martín-Hidalgo1, Mercedes Izquierdo2, Paloma Bartolomé-García3
1Departamento de Bioquímica y Biología Molecular y Genética, Grupo de investigación Señalización Intracelular y Tecnología de la Reproducción (SINTREP), Facultad de Veterinaria, Instituto de Investigación INBIO G+C, Universidad de Extremadura, Av. de las Ciencias, s/n, Cáceres, 10004, Spain.
Abstract:
We aimed to investigate the impact of processing boar spermatozoa with phosphate-buffered saline (PBS) at 4 ˚C on acrosomal integrity and increase in 32 kDa tyrosine-phosphorylated protein (p32). Following cooled PBS washing, we observed a significant increase in p32 levels and in the proportion of dead spermatozoa with compromised acrosomal integrity compared to sperm washing using PBS at room temperature. Interestingly, this increase in p32 was effectively inhibited when cooled PBS was supplemented with 1 mM AEBSF, a serine protease inhibitor. Our findings suggest that the increase of p32 in response to cooled PBS washing in boar spermatozoa is associated with enhanced protease activity in dead spermatozoa.
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