Sanoguet-Crespo, Edith N.

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    Cambios en parametros de motilidad en espermatozoides bovinos inhibiedo diferentes vias metabolicas durante la capacitacion
    (2018) Sanoguet-Crespo, Edith N.; Arcelay-Ruíz, Enid; College of Agricultural Sciences; Sánchez Rodríguez, Héctor; Curbelo Rodríguez, Jaime; Department of Animal Science; Santos Flores,Carlos J.
    The production of beef cattle in Puerto Rico has remained low during the last fifteen years; meanwhile, the demand of this product remains high, depending greatly on importations. The decline in the number of cattle and beef production in the country is notable. Performing assisted reproduction techniques is a strategy to improve local production, so it is essential to know the physiology of the sperm cell. The present study evaluated the mitochondrial function of bovine spermatozoa during in vitro capacitation by using enzymatic inhibitors [(IE); 3-chloro-1,2-propanediol (3MCPD), Sodium Oxamate (SOXAM), 1-fluoro-2,4-dinitrobenzene (DNFB)] and electron transport chain uncouplers [(DES); Rotenone (Rot) and Carbonyl cyanide 3-chlorophenylhydrazone (CCCP)]. Control treatments [capacitating medium (MC) and MC+dimethylsulfoxide (MC+DMSO)] were used to compare the effect with a non-capacitating medium (MNC). Semen from four Senepol bulls was collected by electroeyaculation, incubated for 4 h at 37° C in a 5% CO2 atmosphere, and analyzed (1x/Hr) using the Computer Assisted Sperm Analysis (CASA) system to evaluate the following parameters: motility (MOT), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), beat-cross frequency (BCF), and linearity (LIN). Data were analyzed using the Proc Mixed in SAS (Statistical Analysis System). There was no interaction between treatment and time for the motility-related parameters evaluated. Although, a treatment effect for MOT, VCL, ALH, BCF and LIN (P<0.05) was observed. Similarly, an effect of Hr (P<0.05) was found. Values for BCF increased, while values for MOT, VCL and ALH decreased, indicating sperm hyperactivation through time during the capacitation; for LIN, the values did not demonstrate a pattern. The established controls, MC and MC+DMSO, did not show differences between themselves for any of the motility parameters (P>0.05). Although, between these and the MNC there was a difference for all the parameters (P<0.0001), except for LIN (P=0.6999). The sperm were negatively affected according to the hours of incubation regardless of the treatment: MOT, VCL, ALH, BCF and LIN (P<0.05). On the other hand, using IE vs. MC had no effect for MOT and ALH (P=0.0829 and P=0.1865, respectively); there were differences (P<0.05) however, for VCL, BCF and LIN. For VCL, SOXAM had greater values than MC (P=0.0299) and 3MCPD (P=0.0168); the later was also lower than DNFB (P=0.0364). There was no difference for VCL between MC vs. 3MCPD and DNF (P=0.9977 and P=0.0612, respectively). Even so, MC was less than 3MCPD for BCF (P=0.0583). Sperm treated with SOXAM and DNFB showed a higher percentage of linearity than MC (P=0.0014 and P=0.0443, respectively) and 3MCPD (P=0.0017 and P=0.0513, respectively). These IE caused a significant decrease for MOT, VCL, and ALH between the 1-4 hours of incubation with a difference of 25.19%, 48.51μm/sec, 1.81μm, respectively (P<0.0001). However, BCF increased 4.57Hz (P<0.0001) and LIN was not relevant (P=0.3388). When using DES, compared to MC+DMSO, CCCP reduced 7.61% on MOT (P=0.0115) and 2.55% on LIN (P=0.0075). On the other hand, VCL decreased 13.84μm/sec when the sperms were treated with ROT (P=0.0071). Sperms exposed to ROT and CCCP showed no differences in the parameters of ALH and BCF (P=0.1860 and P=0.3388, respectively). The uncouplers had a significant impact over time causing a decrease in MOT, VCL, and ALH between the 1-4 hours of incubation with a difference of 24.33%, 41.29μm/sec and 1.56μm, respectively (P<0.0001). On the other hand, BCF increased 5.45Hz (P=0.0004) and LIN did not show difference (P=0.4793). These results suggest that bovine sperms can maintain their motility in both conditions, aerobic and anaerobic; although they confirm that their main metabolic pathway is cellular respiration, as long as they have available glycosoluble substrate. Finally, a good management of the sperm improves its fertilizing capacity, probably helping with reduced fertility problems in cattle. This strategy could contribute to improve reproductive performance, and in this way, promote the local production and consumption of beef, which may help the country's economy.