Prion protein 2 (dublet) gene (PRND): role in ovine semen capacitation, cryopreservation and fertility
L. M. Ferreira A , M. Garcia-Herreros B , A. Domingos C , C. C. Marques A , P. Mesquita A D , J. P. Barbas A D , M. C. Baptista A , J. Pimenta A D , A. E. M. Horta A , J. A. M. Prates D and R. M. L. N. Pereira A D E
+ Author Affiliations
- Author Affiliations
A Unidade de Biotecnologia e Recursos Genéticos, Instituto Nacional de Investigação Agrária e Veterinária, Quinta da Fonte Boa, 2005-048 Vale de Santarém, Portugal.
B National Secretariat of Higher Education, Science, Technology, and Innovation (SENESCYT), Av. 9 de Octubre N22-64 and Ramírez Dávalos, PC 170517 Quito, Ecuador.
C Global Health and Tropical Medicine, Instituto de Higiene e Medicina Tropical, Centro de Malária e Doenças Tropicais, Rua da Junqueira 100, 1349-008 Lisboa, Portugal.
D CIISA (Centro de Investigação Interdisciplinar em Sanidade Animal), Faculdade de Medicina Veterinária, Universidade de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal.
E Corresponding author. Email: rosalnp@gmail.com
Reproduction, Fertility and Development 29(5) 985-997 https://doi.org/10.1071/RD15214
Submitted: 18 October 2014 Accepted: 24 January 2016 Published: 6 April 2016
Abstract
The aim of the present study was to examine the role of Doppel protein in the capacitation process and fertilising ability of both fresh and frozen–thawed (FT) spermatozoa from rams carrying different prion protein 2 (dublet) (PRND) gene polymorphisms. The detection efficacy of new anti-Doppel monoclonal antibodies and PRND mRNA quantification were also explored in ovine spermatozoa. Three different genotypes (AA, GA, GG) were identified for codon 26 of ovine PRND-c.78G>A. Using flow cytometry, a higher fluorescence was detected in fresh compared with FT sperm samples incubated with anti-Doppel primary and fluorescein isothiocyanate-conjugated secondary antibodies (P < 0.05). Capacitation was affected by semen treatment (fresh and FT) and male PRND genotype (P < 0.05). After IVF, the use of fresh semen resulted in a higher cleavage rate than the use of FT spermatozoa (P = 0.004). IVF using spermatozoa from individuals classified as carriers of the AA or GA PRND genotypes resulted in higher cleavage rates than seen using spermatozoa from GG carriers (P ≤ 0.0006). Finally, using semen from rams with the AA PRND genotype resulted in the highest Day 6 and Day 8 embryo rates (P ≤ 0.04). In conclusion, the results of the present study confirm that the identification of different PRND genotypes is important for studying the sperm capacitation process and for improving sperm cryoresistance and embryo production. Furthermore, the detection of Doppel in ejaculated ovine spermatozoa, along with its low expression after cryopreservation, strongly suggests an important physiological function of this protein in male fertility.
Additional keywords: Doppel protein, IVF, ovine, sperm capacitation, sperm cryopreservation.
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