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Published on: January 28, 2018
Effect of Mito-TEMPO Incorporated Semen Extender on Physico-morphological Attributes and Functional Membrane
A Kumar1, S K Ghosh2, R Katiyar3
1Division of Animal Reproduction, ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India. abhiawadhesarita@gmail.com.
Background:
Sperm mitochondria are the major site of reactive oxygen species (ROS) production and excess production during freezing-thawing process inflicts oxidative damages to spermatozoa. Buffalo spermatozoa are more prone to oxidative damage due to inherently more polyunsaturated fatty acids and low cholesterol to phospholipids ratio in the plasma membrane. A mitochondrial targeted antioxidant, Mito-TEMPO was used in this study.
Objective:
To study the effect of Mito-TEMPO incorporated semen extender on the post-thaw semen quality in buffalo.
Materials And Methods:
A total of 18 ejaculates from three murrah buffalo bulls with ≥70% individual progressive motility were utilized for the study. Each semen sample was equally divided and extended with five groups: Group I (Control, without Mito-TEMPO addition); Group II (10 µM Mito-TEMPO); Group III (50 µM Mito-TEMPO); Group IV (100 µM Mito-TEMPO); Group V (500 µM Mito-TEMPO) to have 80×106 progressive motile sperm/mL of extender, filled and sealed in French mini straws (0.25 mL) and frozen following equilibration. The effect of Mito-TEMPO was assessed at fresh/post-dilution and post-thaw stages by evaluating physico-morphological attributes and functional membrane integrity such as hypo-osmotic swelling test (HOST).
Results:
Initial progressive motility, viability, acrosomal integrity and HOS response was significantly (p<0.05) improved and sperm abnormality was significantly (p<0.05) reduced in extended semen with Mito-TEMPO (50 µM) compared to control at post-thaw stage, although improvement was also observed at 10 and 100 µM in post-thaw samples.
Conclusion:
Mito-TEMPO incorporated semen extender at 50 µM concentration, could be part of a rationale for improving post-thaw semen quality in buffalo.
Insights
Mito-TEMPO antioxidant supplementation in buffalo semen extenders significantly improved post-thaw sperm quality. The 50 µM concentration of Mito-TEMPO enhanced motility, viability, and membrane integrity, reducing abnormalities in buffalo semen.
Area of Science:
- Reproductive Biology
- Animal Science
- Biochemistry
Background:
- Sperm mitochondria produce reactive oxygen species (ROS), causing oxidative damage during cryopreservation.
- Buffalo spermatozoa are particularly vulnerable to oxidative stress due to membrane composition.
- Mito-TEMPO, a mitochondrial-targeted antioxidant, was investigated to mitigate these damages.
Purpose of the Study:
- To evaluate the efficacy of Mito-TEMPO in semen extenders for improving post-thaw semen quality in buffalo.
- To determine the optimal concentration of Mito-TEMPO for cryopreservation of buffalo spermatozoa.
Main Methods:
- Semen from Murrah buffalo bulls was extended with varying concentrations of Mito-TEMPO (10, 50, 100, 500 µM) or a control.
- Semen quality parameters including progressive motility, viability, acrosomal integrity, and hypo-osmotic swelling test (HOST) were assessed post-thaw.
- Physico-morphological attributes and functional membrane integrity were analyzed.
Main Results:
- A significant improvement (p<0.05) in post-thaw progressive motility, viability, acrosomal integrity, and HOST response was observed with 50 µM Mito-TEMPO.
- Sperm abnormality was significantly reduced (p<0.05) at the 50 µM concentration compared to the control.
- Beneficial effects were also noted at 10 and 100 µM Mito-TEMPO concentrations.
Conclusions:
- Incorporating 50 µM Mito-TEMPO into semen extenders is a promising strategy for enhancing post-thaw semen quality in buffalo.
- Mito-TEMPO supplementation can help protect buffalo spermatozoa from cryodamage.
- This approach offers a rationale for improving artificial insemination success in buffalo.
