Skip to Content
Merck
  • Capacitation and acrosome reaction induction on thawed Dama dama deer spermatozoa: glycine effect as cryopreservation diluent supplement.

Capacitation and acrosome reaction induction on thawed Dama dama deer spermatozoa: glycine effect as cryopreservation diluent supplement.

Zoological science (2013-12-11)
Silvina Fernández, Adrián Sestelo, Miguel Rivolta, Mariana Córdoba
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Glycine 1 M solution
Sigma-Aldrich
Glycine hydrochloride, ≥99% (HPLC)
Supelco
Glycine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Glycine, ReagentPlus®, ≥99% (HPLC)
SAFC
Glycine
Supelco
Glycine hydrochloride solution, 100 mM amino acid in 0.1 M HCl, analytical standard
Sigma-Aldrich
Glycine, 99%, FCC
Sigma-Aldrich
Glycine, meets analytical specification of Ph. Eur., BP, USP, 99-101% (based on anhydrous substance)
Supelco
Glycine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
Glycine, BioUltra, Molecular Biology, ≥99.0% (NT)
Sigma-Aldrich
Glycine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, ≥98.5%
Sigma-Aldrich
Glycine, tested according to Ph. Eur.
Sigma-Aldrich
Glycine, puriss. p.a., reag. Ph. Eur., buffer substance, 99.7-101% (calc. to the dried substance)
Supelco
Glycine, analytical standard, for nitrogen determination according to Kjeldahl method
Sigma-Aldrich
Glycine, BioXtra, ≥99% (titration)
Sigma-Aldrich
Glycine, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Glycine, ACS reagent, ≥98.5%