Fig 1.
Consequences of cryodamage on spermatozoa including structural, functional, and molecular changes.
Created with BioRender.com.
Fig 2.
Motility of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
The values in the columns represent the proportion of motile spermatozoa (%) in each group (±SD). The results were obtained by comparing all groups with each other. The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 3.
Mitochondrial membrane potential of of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
Each bar represents the values of mitochondrial activity (JC1 units) between experimental groups (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 4.
The concentration of cyclic adenosine monophosphate (cAMP) in non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
The graph represented the concentration of cAMP (pmol cAMP/108) between the groups (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 5.
Membrane integrity of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
Each column showed the percentage of cells with intact cell membrane between the groups (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 6.
Acrosome integrity of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
Each bar represented the proportion of spermatozoa (%) with intact acrosome by comparing all groups with each other (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 7.
Occurrence of necrotic spermatozoa.
The graph showed the level of cell necrosis (%) between the fractions of bull spermatozoa (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 8.
DNA fragmentation index of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
The values in the columns represented the percentage (%) of spermatozoa with fragmented DNA between the groups (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 9.
Capacitation CTC patterns of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
All graphs showed individual CTC patterns between groups (±SD). (a) CTC pattern “F”, non-capacitated spermatozoa. (b) CTC pattern “B”, capacitated spermatozoa. (c) CTC pattern “AR”, acrosome-reacted spermatozoa. The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 10.
Intracellular ROS production of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
Each bar represented the concentration of ROS (%) between the groups (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 11.
Intracellular superoxide production of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
The values in columns showed the concentration (%) of superoxide radical between the groups (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 12.
Intracellular production of hydrogen peroxide of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
The graph represented the concentration (%) of hydrogen peroxide in each group (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
Fig 13.
Intracellular production of hydroxyl radical of non-capacitated, in vitro capacitated and cryopreserved bovine spermatozoa.
Each column presents the concentration (%) of hydroxyl radical by comparing all groups with each other (±SD). The level of significance was set at *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.