title: Effects of single layer centrifugation with Androcoll-P on boar sperm.
abstract: Single layer centrifugation (SLC) is a useful technique to select porcine spermatozoa for further artificial insemination practices. The aim of this study was to determine possible side-effects related to capacitation due to the process. Semen viability, acrosome integrity and capacitation status were determined through fluorescent probes (SYBR14-PI, FITC-PSA, CTC staining) and Hsp70 immunolocalization and protein tyrosine phosphorylation (by western blotting and immunolocalization) in different groups: control, after SLC with Androcoll (AND), after SLC and washing (AND-Wash) and after SLC, washing and storage for 2h at 17°C with 2.5% of seminal plasma (AND-Wash-SP). Neither viability nor acrosome integrity were impaired by the different treatments; as far as CTC staining, we observed a significant increase (p<0.05) in the capacitation related pattern in AND and AND-Wash, while after exposure for 2h to seminal plasma (AND-Wash-SP group), the increase became less evident; the same trend was observed in Hsp70 immunolocalization for the EL pattern. Neither immunolocalization nor western blotting for tyrosine phosphorylated proteins had an increase in capacitated pattern or in phosphorylation status, except for a 25kDa band that increased in AND and AND-Wash groups and decreased in AND-Wash-SP group. SLC using Androcoll-P induces some capacitation-related changes in boar sperm membrane, as demonstrated by CTC staining and Hsp70 immunolocalization. For protein tyrosine phosphorylation, only a 25kDa protein showed some changes that should be investigated further.
authors.affiliation: Dipartimento di Scienze Mediche Veterinarie, Università degli Studi di Bologna, Italy. diego.bucci3@unibo.it
authors.forename: D
authors.initials: D
authors.lastname: Bucci
authors.forename: M
authors.initials: M
authors.lastname: Spinaci
authors.forename: J
authors.initials: J
authors.lastname: Morrell
authors.forename: C
authors.initials: C
authors.lastname: Vallorani
authors.forename: C
authors.initials: C
authors.lastname: Tamanini
authors.forename: R
authors.initials: R
authors.lastname: Guidetti
authors.forename: G
authors.initials: G
authors.lastname: Galeati
mesh_terms: D000177:Acrosome / Q000502:physiology; D000818:Animals; D002469:Cell Separation / Q000379:methods* / Q000662:veterinary; D002470:Cell Survival; D002498:Centrifugation / Q000379:methods / Q000662:veterinary; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D007315:Insemination, Artificial / Q000662:veterinary; D008297:Male; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D055101:Semen Analysis / Q000662:veterinary; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D013194:Staining and Labeling / Q000379:methods; D013552:Swine / Q000502:physiology
chemical_list: D018840:HSP70 Heat-Shock Proteins; D011505:Protein-Tyrosine Kinases