Adenosina trifosfato (ATP) na criopreservação de sêmen suíno

Abstract

Pig spermatozoa is a metabolically active cell, which requires energy input, in the form of adenosine triphosphate (ATP), to maintain its homeostasis. From the hydrolysis of ATP in adenosine monophosphate, motor activity is signaled for flagellar movement. However, when the spermatozoa of the swine species are subjected to technological processes of semen conservation, such as cryopreservation, there are energy expenditures above the normally expected, with losses in the concentration of viable cells, promoting low sperm motility reducing their seminal quality. Cellular structures such as plasma membrane, mitochondria, and acrosome may be negatively affected by the cryoinjurias that undergo the cells due to the freeze / thaw processes of cryopreservation. The objective of this study was to evaluate the effects of the addition of exogenous ATP, in different concentrations, in the swine semen freezing diluent, through kinetic analysis and cellular structures. In the cryopreservation methodology, diluents of cooling, Lactose-11% (80%) and egg yolk were used. The study was performed from 23 samples of inseminating doses of porcine ejaculates (n = 23) from an insemination center. (20%) and freezing with 89.5% of DR + 1.5% Equex-Paste® and 9% Glycerol. In the freezing diluent, exogenous ATP (Adenosine 5'-triphosphate disodiumsalthydrate - Sigma-Aldrich (0.25 mM), T4 (2.5 mM) and T5 (25.0 mM). The vals obtained a total volume of 0.5 mL each Concentration of 500x106 spermatozoa, were thawed in a water bath (37 °C) and the kinetics were read in the three-phase Computer Assisted Sperm Analysis (SpermVision 3.5) system and the cell structures by the Flow Cytometer (Attune® Cytometer) The results were varied, however, it was evidenced that the treatment with 0.25 mM of ATP was the most satisfactory (Total time and progressive motility), while the treatment with 25mM was the one that presented the lowest performance, but this one, obtained a better result in the times of one and two hours after the thawing.