99 Short-term storage of equine embryos at 5 or 20°C does not cause lipid peroxidation
| Autor: | D. Scarlet, Christine Aurich, G. D. A. Gastal |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Embryo culture
Embryo Reproductive technology Biology Malondialdehyde Cryopreservation Andrology Lipid peroxidation chemistry.chemical_compound Endocrinology Human fertilization Reproductive Medicine chemistry Genetics Animal Science and Zoology Molecular Biology Gametogenesis Developmental Biology Biotechnology |
| Zdroj: | Reproduction, Fertility and Development. 32:175 |
| ISSN: | 1031-3613 |
| DOI: | 10.1071/rdv32n2ab99 |
| Popis: | Maintaining the integrity of equine embryos during storage for transportation is essential for successful conception after transfer. During storage, reactive oxygen species may originate from embryo metabolism, causing lipid peroxidation and increasing its end product malondialdehyde (MDA); MDA is one of the important biomarkers for oxidative stress. This study aimed to evaluate the temperature curve, pH and lipid peroxidation of equine embryos stored in holding medium after 24h at 5 or 20°C. Embryos (n=33) were collected on Day 7 (n=21, 7 embryos per group) or Day 8 (n=12) after ovulation and assigned to four groups: Day 7 control (D7, fresh); Day 7, 24h at 5°C (E5C); Day 7, 24h at 20°C (E20C); and Day 8 control (D8, fresh 24h time control). After collection, embryos were washed and kept in holding medium (Minitube) for morphological classification and measurements. For pH and lipid peroxidation measurement, embryos were kept in a fixed volume of holding medium (150µL) within a microtube (200µL); the microtube was kept within a falcon tube (50mL) inside of an Equitainer (Hamilton Biovet). The temperature was recorded by a data logger (Testo 175, Testo) every 10min for 24h. The pH was assessed by a pH meter using a microelectrode (InLab Ultra-Micro-ISM, Mettler Toledo) for small sample volumes. Lipid peroxidation was assessed using the MDA assay kit (catalog number MAK085, Sigma-Aldrich) according to the manufacturer's instructions. Statistical analyses were performed using the Kruskal-Wallis nonparametric test and Mann-Whitney U to compare differences among groups. Embryo size differed (P0.05). The temperature curve was similar (P>0.05) among embryos within the treatment groups during the storage period. The pH (7.22±0.07 and 7.22±0.09) did not differ (P>0.05) between E5C and E20C. Lipid peroxidation levels were below the limit of quantification (0.04 nmol) in all groups. The present findings suggest that holding temperature does not affect the size, pH, or lipid peroxidation of equine embryos stored for 24h in holding medium. However, our previous studies (Gastal et al. 2018 J. Equine Vet. Sci. 66, 185) have shown that holding temperature influences the expression of genes involved in equine embryo development. In conclusion, variations regarding embryo development and conception rate after transfer of embryos stored at different temperatures might be related to factors other than lipid peroxidation. |
| Databáze: | OpenAIRE |
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