A novel approach to identifying physical markers of cryo-damage in bull spermatozoa.
Cryopreservation is an efficient way to store spermatozoa and plays a critical role in the livestock industry as well as in clinical practice. During cryopreservation, cryo-stress causes substantial damage to spermatozoa. In present study, the effects of cryo-stress at various cryopreservation steps, such as dilution / cooling, adding cryoprtectant, and freezing were studied in spermatozoa collected from 9 individual bull testes. The motility (%), motion kinematics, capacitation status, mitochondrial activity, and viability of bovine spermatozoa at each step of the cryopreservation process were assessed using computer-assisted sperm analysis, Hoechst 33258/chlortetracycline fluorescence, rhodamine 123 staining, and hypo-osmotic swelling test, respectively. The results demonstrate that the cryopreservation steps reduced motility (%), rapid speed (%), and mitochondrial activity, whereas medium/slow speed (%), and the acrosome reaction were increased (P < 0.05). Differences (Δ) of the acrosome reaction were higher in dilution/cooling step (P < 0.05), whereas differences (Δ) of motility, rapid speed, and non-progressive motility were higher in cryoprotectant and freezing as compared to dilution/cooling (P < 0.05). On the other hand, differences (Δ) of mitochondrial activity, viability, and progressive motility were higher in freezing step (P < 0.05) while the difference (Δ) of the acrosome reaction was higher in dilution/cooling (P < 0.05). Based on these results, we propose that freezing / thawing steps are the most critical in cryopreservation and may provide a logical ground of understanding on the cryo-damage. Moreover, these sperm parameters might be used as physical markers of sperm cryo-damage.
Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do, 456-756, Republic of Korea.
Sung-Jae
SJ
Yoon
Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do, 456-756, Republic of Korea.
Woo-Sung
WS
Kwon
Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do, 456-756, Republic of Korea.
Md Saidur
MS
Rahman
Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do, 456-756, Republic of Korea.
June-Sub
JS
Lee
Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do, 456-756, Republic of Korea.
Myung-Geol
MG
Pang
D020101:Acrosome Reaction; D000818:Animals; D015415:Biomarkers; D002417:Cattle; D015925:Cryopreservation / Q000379:methods; D005434:Flow Cytometry; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D055101:Semen Analysis; D012662:Semen Preservation / Q000379:methods; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology*
D015415:Biomarkers