Seminal plasma improves cryopreservation of Iberian red deer epididymal sperm

Abstract

We tested the protective action of seminal plasma on epididymal spermatozoa from Iberian red deer, especially considering cryopreservation, as a means for germplasm banking improvement. We obtained seminal plasma by centrifuging electroejaculated semen, and part of it was thermically inactivated (denatured plasma; 55 degrees C 30 min). Epididymal samples (always at 5 degrees C) were obtained from genitalia harvested after regulated hunting, and pooled for each assay (five in total). We tested three seminal plasma treatments (mixing seminal plasma with samples 2:1): no plasma, untreated plasma and denatured plasma; and four incubation treatments: 32 degrees C 15 min, 5 degrees C 15 min, 5 degrees C 2h and 5 degrees C 6h. After each incubation, samples were diluted 1:1 with extender: Tes-Tris-Fructose, 10% egg yolk, 4% glycerol; equilibrated for 2h at 5 degrees C, extended down to 10(8) spz./mL and frozen. Sperm quality was evaluated before 1:1 dilution, before freezing and after thawing the samples, assessing motility (CASA) and viability (percentage of viable and acrosome-intact spermatozoa; PI/PNA-FITC and fluorescent microscopy). Plasma treatment, both untreated and denatured, rendered higher viability before freezing and higher results for most parameters after thawing. The improvement was irrespective of incubation treatment, except for viability, which rendered slightly different results for untreated and denatured plasma. This may be due to the presence of thermolabile components. We still have to determine the underlying mechanisms involved in this protection. These results might help to improve the design of cryopreservation extenders for red deer epididymal sperm.

Artificial Insemination in Veterinary Science

“…Epididymal spermatozoa have been harvested from a variety of species like cats (Filliers et al, 2008;Hermansson & AxnÈr, 2007), dogs (Garcia-Macias et al, 2006;Hewitt et al, 2001;Nothling et al, 2007;Ponglowhapan et al, 2006), rats (Yamashiro et al, 2007), horses (Braun et al, 1994;Bruemmer, 2006;Heise et al, 2011;Johnson & Coutinho da Silva, 2008;Melo et al, 2008), cattle (Goovaerts et al, 2006;Martins, Rumpf, Pereira, & Dode, 2007), pigs (Ikeda et al, 2002), sheep (Garcia-Macias et al, 2006), goats (Blash, Melican, & Gavin, 2000), red deer (Fernandez-Santos et al, 2006;Garcia-Macias et al, 2006;MartÌnez-Pastor et al, 2006), Spanish Ibex , African buffalo (Herold et al, 2006), North American buffalo (Lessard at al., 2009) and monkeys (Goff et al, 2009;Ng et al, 2002). Pregnancies and offspring after AI with epididymal spermatozoa have been produced amongst others in horses (Barker & Gandier, 1957;Heise et al, 2010;Papa et al, 2008), dogs (Hori, Hagiuda, Kawakami, & Tsutsui, 2005) and Spanish Ibex (SantiagoMoreno et al, 2006).…”

Effects on brown bear (Ursus arctos) spermatozoa freezability of different extender and dilution ratios used for pre-freezing centrifugation

“…A consequence of centrifugation is the removal of seminal plasma and their potential effects on semen quality. Although some studies in several species have shown the beneficial effects of seminal plasma addition on spermatozoa during processing and cryopreservation (deer: Martinez-Pastor et al 2006;ram: Ollero et al 1997), others have reported deleterious effects (dog: Rota et al 2006;goat: Pellicer-Rubio et al 1997;ram: Ritar and Salamon 1982;stallion: Carver and Ball 2002;Sieme et al 2004;bull: Way et al 2000).…”

The Quality of Cryopreserved Sperm Collected from Feline Caudal Epididymides Using Seminal Plasma

“…In other animal species, improvement in sperm qualities or fertility by SP has been frequently reported [3,4,6,7]. For example, in Iberian red deer [6], no difference due to the presence or absence of SP was noted in epididymal sperm motility before freezing, but motility was significantly higher when sperm were treated with SP after freezing-thawing. The same phenomenon has been reported [4,7] in dogs.…”

“…However, no difference was noted in lectin binding between feline epididymal and ejaculated sperm. The effects of SP on sperm functions have been reported to be positive or negative in various animal species [3,4,[6][7][8]11].Kawakami et al[5] investigated protein attachment on the surfaces of canine caudal epididymal and ejaculated sperm. They observed that PHA-E lectin did not bind to caudal epididymal sperm but bound to ejaculated sperm and suggested that a PHA-E lectin-binding glycoprotein, N-acetyl-D-galactosamine, is a decapacitation factor.…”

“…However, no difference was noted in lectin binding between feline epididymal and ejaculated sperm. The effects of SP on sperm functions have been reported to be positive or negative in various animal species [3,4,[6][7][8]11].…”