Intracellular calcium chelating agent (BAPTA-AM) aids stallion semen cooling and freezing–thawing

Shuaishuai Wu, Igor F. Canisso, Weigang Yang, Ihteshamu Ul Haq, Qiang Liu, Ying Han, Shenming Zeng

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish (US)
Pages (from-to)1235-1242
Number of pages8
JournalReproduction in Domestic Animals
Issue number5
StatePublished - Oct 2018


  • horse
  • semen cooling
  • semen cryopreservation
  • sperm quality

ASJC Scopus subject areas

  • Biotechnology
  • Animal Science and Zoology
  • Endocrinology


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