title: PSP-I/PSP-II spermadhesin exert a decapacitation effect on highly extended boar spermatozoa.
abstract: PSP-I/PSP-II heterodimer is a major protein of boar seminal plasma that is able to preserve, in vitro, the viability, motility and mitochondrial activity of highly-extended boar spermatozoa. However, a relationship between the protective effects of the heterodimer and sperm capacitation is still unclear. The present study investigated the effect of the PSP-I/PSP-II (1.5 mg/mL) on membrane stability, intracellular calcium concentration ([Ca(2+)](I)) and plasma membrane and acrosome integrity of highly extended boar spermatozoa. Boar spermatozoa were diluted to 1 x 10(6) spermatozoa/mL and incubated at 38 degrees C in Phosphate-buffered saline (PBS) for 10, 30, 60, 120 and 300 min or in modified Tris-buffered medium (mTBM) for 10, 20, 30, 60 and 120 min. After each incubation time, the membrane stability (using Merocyanine-540/Yo-Pro-1), elevation of [Ca(2+)](I) (using Fluo-3-AM/PI) and the sperm plasma membrane and acrosome integrity (using SYBR-14/PI/PE-PNA) were evaluated by flow cytometry. As expected, exposure of the spermatozoa to the PSP-I/PSP-II preserved the plasma membrane and acrosome integrity compared to non-exposed spermatozoa in both media PBS and mTBM (p < .01). The evaluation of membrane stability showed no differences in the percentages of viable sperm with instable plasma membrane in the presence of the PSP-I/PSP-II compared to controls irrespective of the dilution media. The evaluation of the [Ca(2+)](I) levels showed that while spermatozoa incubated in mTBM and exposed to PSP-I/PSP-II had lower [Ca(2+)](I) than controls (39.08% vs. 47.97%, respectively; p < .05), no differences were observed in those samples incubated in PBS. However, a temporal evaluation of the samples showed that a similar proportion of live spermatozoa were able to achieve high levels of [Ca(2+)](I) and membrane instability independent of the presence of PSP-I/PSP-II. In conclusion, PSP-I/PSP-II exert a non-permanent decapacitation effect on highly extended boar spermatozoa that is related with a delay in the increase of [Ca(2+)](I) levels.
authors.affiliation: Department of Medicine and Surgery, Faculty of Veterinary Medicine, University of Murcia, Murcia, Spain. nachof@um.es
authors.forename: Ignacio
authors.initials: I
authors.lastname: Caballero
authors.forename: Juan M
authors.initials: JM
authors.lastname: Vazquez
authors.forename: Gloria M
authors.initials: GM
authors.lastname: Mayor
authors.forename: Carmen
authors.initials: C
authors.lastname: Almiñana
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Calvete
authors.forename: Libia
authors.initials: L
authors.lastname: Sanz
authors.forename: Jordi
authors.initials: J
authors.lastname: Roca
authors.forename: Emilio A
authors.initials: EA
authors.lastname: Martinez
mesh_terms: D000818:Animals; D019281:Dimerization; D005434:Flow Cytometry; D008297:Male; D029589:Seminal Plasma Proteins / Q000502:physiology*; D029607:Seminal Vesicle Secretory Proteins / Q000502:physiology*; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000166:cytology; D013552:Swine
chemical_list: D029589:Seminal Plasma Proteins; D029607:Seminal Vesicle Secretory Proteins; C425042:spermadhesin; C074520:seminal vesicle secretory protein 109, porcine