TY - JOUR
T1 - Intracellular calcium chelating agent (BAPTA-AM) aids stallion semen cooling and freezing–thawing
AU - Wu, Shuaishuai
AU - Canisso, Igor F.
AU - Yang, Weigang
AU - Ul Haq, Ihteshamu
AU - Liu, Qiang
AU - Han, Ying
AU - Zeng, Shenming
N1 - Funding information This work was supported by the Specialized Research Fund for the Doctoral Program for Higher Education (20130008130001) and the Natural Science Foundation of China (31470077). The authors are grateful for the providential assistance provided by all the clinicians and supporting staff at the Equine Teaching and Research Center of the China Agricultural University.
This work was supported by the Specialized Research Fund for the Doctoral Program for Higher Education (20130008130001) and the Natural Science Foundation of China (31470077).
PY - 2018/10
Y1 - 2018/10
N2 - 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.
AB - 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.
KW - horse
KW - semen cooling
KW - semen cryopreservation
KW - sperm quality
UR - http://www.scopus.com/inward/record.url?scp=85050502241&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85050502241&partnerID=8YFLogxK
U2 - 10.1111/rda.13245
DO - 10.1111/rda.13245
M3 - Article
C2 - 29984866
AN - SCOPUS:85050502241
SN - 0936-6768
VL - 53
SP - 1235
EP - 1242
JO - Reproduction in Domestic Animals
JF - Reproduction in Domestic Animals
IS - 5
ER -