Intracellular calcium chelating agent (BAPTA‐AM) aids stallion semen cooling and freezing–thawing. (9th July 2018)

• Record Type: Journal Article
• Authors: Wu, Shuaishuai; Canisso, Igor F.; Yang, Weigang; Ul Haq, Ihteshamu; Liu, Qiang; Han, Ying; Zeng, Shenming
• Is Part Of: Reproduction in domestic animals. Volume 53:Number 5(2018)
• Page Start: 1235
• Page End: 1242
• Publication Date: 2018-07-09
• Subjects: horse -- semen cooling -- semen cryopreservation -- sperm quality; Animal breeding -- Periodicals; Veterinary pathology -- Periodicals; Livestock -- Breeding -- Periodicals; 636.082
• DOI: 10.1111/rda.13245
• Languages: English
• ISSNs: 0936-6768

Abstract

Abstract: This study aimed to investigate the effects of different concentrations of 1, 2‐bis‐(o‐aminophenoxy)‐ethane‐N, N, N0 N0‐tetraacetic acid, tetra‐acetoxymethyl ester (BAPTA‐AM), an intracellular calcium chelating agent, on stallion semen cooling and freezing–thawing. After collection, semen was extended (1:1 v/v) on a skim milk‐based extender, centrifuged and resuspended at 400 million/ml into cooling or freezing extenders containing 0, 5, 25, 50, 100 and 200 μΜ BAPTA‐AM. Motility parameters were assessed after cooling in Equitainer at 5°C for 12, 24, 48, 72 and 120 hr and after freezing–thawing. In addition, mitochondrial membrane potential, intracellular ATP, reactive oxygen species and malondialdehyde concentrations were measured in cryopreserved‐thawed semen. Cooled stored (48 hr) semen containing 50 μΜ BAPTA‐AM and control extender (0 μΜ BAPTA‐AM) was used to assess fertility. Inclusion of 50 μΜ BAPTA‐AM resulted in superior sperm motility parameters during cooled storage when compared to other groups ( p < 0.05). Furthermore, semen cryopreserved in extender containing 50 μΜ BAPTA‐AM showed increased intracellular ATP and mitochondrial membrane potential, whereas reactive oxygen species and malondialdehyde were increased after thawing for all groups ( p < 0.05). Addition of 50 μΜ BAPTA‐AM to cooling extender resulted in similar pregnancy rates to the control group (75% vs. 73.6%, respectively; p > 0.05). In conclusion, the addition of BAPTA‐AM to semen extenders aided stallion semen cryopreservation in a dose‐dependent manner. Furthermore, the cooling extender supplemented with 50 μΜ BAPTA‐AM could be used to prolong the sperm motility during cooling without apparently compromising fertility. Field trials should be conducted to assess fertility of cryopreserved stallion semen with BAPTA‐AM.