Novel droplet vitrification combined with fish antifreeze protein type III enhances cryoprotection of semen in wild endangered Persian sturgeonAcipenser persicus(Borodin, 1897)

• DOI: 10.1111/are.12397
• Publication Date: 2014-02-01T06:26:41Z
• Authors: Amirreza Abed-Elm...; Hamid Farahmand; Bagher Mojazi-Amiri; Gholamreza Rafiee; Ruhollah Rahimi
• Keywords: 0301 basic medicine; 03 medical and health sciences

AbstractIn this study, the efficiency of a novel dropletvitrification technique along with different dosesof fish antifreeze protein (AFP) type III on Per-sian sturgeon thawed spermatozoa quality (motil-ity duration and motility percentage) wasinvestigated. Semen of seven male individualswas pooled in equal volumes and diluted with4°C Tris-Hcl (100 mM), pH = 8 extenders con-taining 0, 5, 10, 15 lM of AFP type III in aratio of 1:1 (semen/extenders). Treated semenwas dropped into liquid nitrogen. Solidified drop-lets were stored for 2, 60 and 120 days andthawed by plunging them into a tube containing5 mL Tris-Hcl (100 mM), pH=8 with 1% BSA at37°C. Motility duration in all treatments had nosignificant difference comparing to fresh sperm(P > 0.05), but their motility percentage wassignificantly lower. Treatment with 10 lMofAFP had significantly higher motility percentage(16.11 0.5%) comparing to other treatments(P 0.05), suggesting that antifreezeprotein effectiveness are highly dose dependent,and dose of 10 lM is appropriate in Persiansturgeon spermatozoa droplet vitrification.Besides, the present technique obtained higherquality of spermatozoa comparing to its analoguetechniques.Keywords: Droplet vitrification, Wild Persionsturgeon semen, Fish antifreeze protein type IIIIntroductionThe Persian sturgeon is endemic to the Caspianand Black Sea basins. They are commerciallyimportant fish valued for their meat, but mainlyfor their caviar (Alavi, Cosson, Karami, Mojazi A the first is termed traditional, controlledrate, colligative or conventional freezing and thesecond is called vitrification (Day, Harding,Nadarajan & Benson 2008; Xu, Wang, Wu, Meng,Wu, Zhou & Zhou 2012).On controlled slow cooling, extracellular ice for-mation and vapour discrepancy created across thecell membrane cause the water leave the cell andpenetration of colligative cryoprotectants protectsthe cells against the toxic effects of solute concen-tration (Day et al. 2008). On the other hand, vitri-fication is the rapid cooling of liquid medium inthe absence of ice crystal formation (Chao & Liao2001) and curtails the physical injuries to the cell;In contrast to slow (programmed) cooling, the pro-tocols for vitrification are simple and require only