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Dissecting the Protective Effect of the Seminal Plasma Spermadhesin PSP-I/PSP-II on Boar Sperm Functionality

JUAN J. CALVETE

LIBIA SANZ

From the ^* Departamento de Medicina y Cirugía Animal, Facultad de Veterinaria, Universidad de Murcia, Murcia, Spain; and the Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain.

Correspondence to: Juan M. Vázquez, Departamento de Medicina y Cirugía Animal, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, 30071 Murcia, Spain (e-mail: vazquez{at}um.es).

To dissect the protective activity of PSP-I/PSP-II, the effect of the isolated subunits PSP-I and PSP-II and their affinity-purified tryptic peptide and glycan fractions on the viability, mitochondrial activity, and motility of highly diluted boar spermatozoa was investigated. High dilution exerted a negative effect on control spermatozoa. Incubation of spermatozoa with PSP-I/PSP-II or with its PSP-II subunit had a protective effect on sperm functionality, high mitochondrial membrane potential, and sperm motility. These effects were less pronounced when spermatozoa were incubated with the PSP-I subunit. It was noteworthy that motility was abolished by incubation of spermatozoa with isolated PSP-I. Trypsin-degraded PSP-I/PSP-II, PSP-I, and PSP-II reproduced the effects of the native proteins. Incubating spermatozoa with the glycan-depleted tryptic-peptide fraction of PSP-I/PSP-II for 5 hours preserved a higher percentage of viable spermatozoa than when sperm was incubated for the same time with the native heterodimer, trypsin-digested PSP-I/PSP-II, the glycan fraction or without added proteins. However, sperm motility decreased as the concentration of added peptide fraction increased. On the other hand, spermatozoa incubated with the glycan fraction showed lower values than spermatozoa incubated with the peptide fraction. We concluded that the subunits of the PSP-I/PSP-II heterodimeric spermadhesin exert different activities on sperm functions. The finding that the beneficial effect of the native PSP-I/PSP-II on the functionality of highly diluted boar spermatozoa is largely preserved in its isolated PSP-II subunit and does not appear to require the glycan moiety points to a peptide moiety as a potential sperm function-preserving additive of highly diluted boar spermatozoa.

     Key words: Boar seminal plasma, spermadhesin PSP-I/PSP-II, protective effect on sperm function, sperm survival, highly diluted spermatozoa

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