Capacitation and Acrosome Reaction Induction on ThawedDama damaDeer Spermatozoa: Glycine Effect as Cryopreservation Diluent Supplement

Abstract

The aim of this research was to evaluate two different diluents for sperm cryopreservation and to study functional parameters in relation to the response to heparin, lysophosphatidylcholine and progesterone, in frozen-thawed semen of fallow deer (Dama dama) during the reproductive season (brama). In this way, fallow deer can be used as a biological model of endangered cervids. Semen was obtained by electroejaculation. Heparin, progesterone and lysophosphatidylcholine were used as capacitation and acrosome reaction inducers, respectively. Capacitation and acrosome reaction were evaluated by chlorotetracycline epifluorescence technique (CTC), membrane integrity by Hypo-osmotic swelling test (HOS) and viability and acrosome integrity by trypan blue stain/DIC. Data was analyzed by ANOVA and Tukey Test (P < 0.05). Semen was cryopreserved in different diluents and Fructose-Tris-Glycine extender was selected. Capacitation with heparin at different incubation times determined that the highest capacitation percentage was obtained at 45 minutes incubation. Progesterone (1 'M) and lysophosphatidylcholine in heparin capacitated sperm induced acrosome reaction (P < 0.05). This study contributes to improve cryopreservation methods and to increase the knowledge about capacitation and acrosome reaction in vitro in deer spermatozoa, allowing an advance in the development of reproductive biotechnologies.

Sperm characterization and identification of sperm sub-populations in ejaculates from pampas deer (Ozotoceros bezoarticus)

“…Thus, it is essential the cryopreservation of pampas deer sperm to implement reproductive biotechnologies. As deer are not easily domesticated, sperm are usually obtained by electroejaculation, as occurs in other deer species (Fernández et al, 2013;Jacobson et al, 1989).…”

Improvement in non-programmable sperm cryopreservation technique in farmed greenlip abalone Haliotis laevigata

“…This layer in turn can subsequently influence the permeability of DMSO entering into sperm, resulting in the DMSO osmotic or toxic stress being minimized (Anchordoguy et al, 1988;He and Woods, 2004). The benefit of glycine has been also found in Dama dama deer (Fernández et al, 2013), and grayling Thymallus thymallus (Lahnsteiner et al, 1992). No significant difference in post-thaw sperm fertilization rate was found with the addition of glucose, galactose or fructose in 6% DMSO, although the addition of glucose resulted in a higher numerical rate than the other two treatments.…”

Characterization of seminal parameters, sperm morphometry, micromorphology, and ultrastructure in gray brocket deer (Mazama gouazoubira, Fischer, 1814)

“…The total mean percentage of viable cells recorded in this study (73.4 ± 18.3) was lower than that recorded for O. bezoarticus (85 to 96%) (Beracochea et al, 2014). For thawed cryopreserved samples of C. elaphus hispanicus and in D. dama , the mean percentage of viable cells was 27% (Martínez‐Pastor et al, 2006) and 82.37% (Fernández et al, 2013), respectively. Non‐reactive cells on HOST might have suffered problems during the maturation phase, developing a less effective capacity to control volume.…”