Animal Reproduction Science 138 (2013) 276–281 Contents lists available at SciVerse ScienceDirect Animal Reproduction Science journal h om epa ge: www.elsevier.com/locate/anireprosci Effects of single layer centrifugation with Androcoll-P on boar sperm D. Bucci a,∗ , M. Spinaci a , J. Morrell b , C. Vallorani a , C. Tamanini a , R. Guidetti c , G. Galeati a a Dipartimento di Scienze Mediche Veterinarie (DSMVet), Università degli Studi di Bologna, Italy b Swedish University of Agricultural Sciences, Uppsala, Sweden c Inseme S.p.a. Modena, Italy a r t i c l e i n f o Article history: Received 6 August 2012 Received in revised form 20 February 2013 Accepted 28 February 2013 Available online 14 March 2013 Keywords: SLC Androcoll-P Capacitation-related changes a b s t r a c t 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 2 h 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 2 h 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 25 kDa 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 mem- brane, as demonstrated by CTC staining and Hsp70 immunolocalization. For protein tyrosine phosphorylation, only a 25 kDa protein showed some changes that should be investigated further. © 2013 Elsevier B.V. All rights reserved. 1. Introduction Density gradient centrifugation has been used to select spermatozoa for IVF in most of the domestic species, as well as in human reproduction (Mortimer, 2000; Morrell, 2006). However, the limit of this procedure is represented by the low volume of semen that can be processed each ∗ Corresponding author at: Via Tolara di Sopra, 50, 40064 Ozzano dell’Emilia, Bologna, Italy. Tel.: +39 0512097904; fax: +39 0512097899. E-mail addresses: diego.bucci3@unibo.it, diego.bucci1@gmail.com (D. Bucci). time. Recently, a new technique, Single Layer Centrifu- gation (SLC) (Sharma et al., 1997; Morrell, 2006; Morrell et al., 2009) has been set up and introduced and impor- tant improvement of this new technique is the substitution of Percoll, considered potentially dangerous to sperm cells (Mortimer, 2000) with species-specific colloids, based on silane-coated silica, to boar (Morrell et al., 2009), horse (Macías García et al., 2009a,b) and bull (Thys et al., 2009) spermatozoa. Focusing on porcine species, the Swedish University of Agricultural Science (SLU) has recently developed a col- loid, Androcoll-P (Morrell et al., 2009) to be used in the SLC technique (Morrell and Wallgren, 2011). This application 0378-4320/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.anireprosci.2013.02.020