55 DIFFERENTIAL EXPRESSION OF AQUAPORINS AND SPERMADHESINS IN FROZEN–THAWED 'GOOD FREEZER' AND 'POOR FREEZER' BOAR SPERMATOZOA

• DOI: 10.1071/rdv26n1ab55
• Publication Date: 2013-12-05T01:20:15Z
• Authors: A. C. M. Filho; R. M. Brezinsky; R. C. Youngblood; L. D. M. Da Silva; S. T. Willard; P. L. Ryan; J. M. Feugang
• Keywords: BOAR; Capacitation; Percoll

Cryopreservation is an excellent tool for sperm banking in livestock animals. Although its utilisation widespread species such as cattle, application pigs limited due to various factors associated with either cryopreservation techniques or male-to-male variations. Indeed, the existence of so-called “good freezer” (or GF) and “poor (PF) males has been well established swine yet, potential screen these characteristics still being investigated. Few studies have reported molecular differences between both groups boar spermatozoa, at nucleic acid protein levels. Here we hypothesise that cryotolerance may be differential expression proteins involved membrane transport capacitation. Thus, conducted this study compare spermadhesins, aquaporins (AQP), glucose (GLUT) transporters GF PF frozen-thawed spermatozoa. Three ejaculates 8 boars classified (n = 4) 4), based on post-thaw fertility records were used. Frozen-thawed spermatozoa purified using a Percoll gradient, washed 3 times PBS/polyvinylpyrrolidone (PVP). Total RNA extraction took place different purifications each sire testis cells, was followed by cDNA synthesis. Primer sets designed detection AQP isoforms 1 11, GLUT-3 GLUT-5, spermadhesins (AQN-1, AWN-1, SPM-I, PSP-1, PSP-2) used real-time PCR (pool ejaculates/boar), GAPDH serving housekeeping gene, cells positive controls. Gene analysed comparative quantitation (REST 2009 software; Qiagen Inc., Valencia, CA) via Pfaffl method, amplicons verified gel electrophoreses. The water (AQP-1, -5, -11) glycerol (AQP-3, -7, -9, -10) transmembrane transporters, detected all samples. Expression levels AQP-10 AQP-11 greater than other tested (P < 0.05), no expressions found (GF v. PF; P > 0.05). transcript lower versus (0.705×; Irrespective groups, AQN AWN highest lowest, respectively PSP-1 mRNA level reduced (0.431×; whereas comparable findings indicate express transporter transcripts are essential fluxes occurring during cryopreservation. Additionally, spermadhesin raises questions about their roles early capacitation Differential genes brings additional evidence contribute better description from Further investigations undergoing full characterisation Supported US Department Agriculture-Agricultural Research Service (USDA-ARS) Grant #58-6402-3-0120 Coordenação de Aperfeiçoamento Pessoal Nível Superior (CAPES)-Brazil.