Effects of Leucosporidium‐derived ice‐binding protein (LeIBP) on bull semen cryopreservation

Abstract

We examined the effect of ice‐binding protein derived from Leucosporidium (LeIBP) on the cryopreservation of bull semen and compared it with that derived from previously reported Antifreeze Protein III (AFPIII). Six concentrations of LeIBP (10 –1 ~ 10 4 μg/ml) and AFPIII (10 –1 ~ 10 4 μg/ml) were added to the bull semen extender, respectively. Sperm kinematic parameters were measured to examine sperm toxicity and cryopreserved sperm quality. Measures of antioxidant activity such as superoxide dismutase (SOD), reduced glutathione/oxidative glutathione (GSH/GSSG), and total antioxidant capacity (TAC) were analysed to identify the effect of LeIBP on sperm quality. In addition, sperm viability was analysed using a flow cytometer and fluorescence microscope by SYBR14/PI staining. The results showed that the LeIBP groups (0.1, 1 and 10 μg/ml) were less toxic, and the quality of the sperm were dramatically improved in the extenders containing 0.1 μg/ml LeIBP among concentrations of LeIBP and AFPIII. The SOD activity of LeIBP was greater than that of AFPIII and control. In addition, sperm viability was enhanced in the LeIBP‐treated group. In summary, LeIBP is a useful cryoprotective adjuvant for bull sperm cryopreservation, and the most efficient concentration of LeIBP is 0.1 μg/ml.

Improving Sperm Cryopreservation With Type III Antifreeze Protein: Proteomic Profiling of Cynomolgus Macaque (Macaca fascicularis) Sperm

“…Furthermore, ROS level was decreased in mouse oocytes vitrified with AFP III (Salvay et al, 2010 ). Another study also demonstrated that reduced glutathione/oxidative glutathione (GSH/GSSG) and total antioxidant capacity (TAC) were higher in bull semen cryopreservation with AFP III compared with the control group (Jang et al, 2020 ). In our study, the results also suggested that AFP III could protect the proteins associated with ROS production in mitochondria and GSH synthesis during sperm freezing and thawing.…”

Antifreeze Proteins: Novel Applications and Navigation towards Their Clinical Application in Cryobanking

“…AFP III improved survival by preserving protein expression and modulating the release of cytochrome C and free radicals at the molecular level. Jang et al [ 98 ] compared LeIBP with AFP III in the preservation of Korean bull semen and LeIBP showed better outcomes even in embryo formation after IVF.…”

Application of Nanoparticles and Melatonin for Cryopreservation of Gametes and Embryos

“…In particular, embryos are very difficult to cryopreserve because of their size, survival sensitivity, and multi-compositional structure [ 7 , 8 ]. Several studies have reported that aberrant effects such as genotoxicity, necrosis, and abnormal apoptotic activity are responsible for the cytotoxicity observed in gametes and embryos during cryopreservation and vitrification [ 9 , 10 , 11 , 12 ]. To overcome these impediments, a number of researchers have studied the effect of co-treatment of gametes and embryos with protective adjuvants and freezing media, which was found to reduce toxicity as well as increase the success rate of artificial insemination.…”

“…Several organisms that live in extremely cold temperatures, including bacteria, fungi, microalgae, insects, crustaceans, and fish, produce specific proteins, called antifreeze proteins (AFPs) or ice-binding proteins (IBPs), that can inhibit ice recrystallization [ 9 , 11 , 13 , 14 , 15 , 16 , 17 , 18 , 19 ]. In a subzero temperature environment, these proteins interact with ice crystals and regulate thermal hysteresis in order to stabilize the cell membrane [ 20 ].…”