Plasma membrane, acrosome and mitochondria analysis of sheep sperm after cryopreservation with addition of folic acid in the tris-yolk medium
Keywords:
Folic acid, Sheep, SemenAbstract
Folic acid is closely linked to cobalamin, which is a carrier of hydroxymethyl and ant groups. The objective of this work was to evaluate the effect of adding folic acid to the TRIS-yolk diluter and possible effect on the membrane, mitochondria and acrosome of sheep sperm after thawing the semen. There were six sheep and seven collections of each animal in sessions between 48 and 72 hours. After collection and analysis, the semen samples were mixed and submitted to the formation of a pool. Diluted in TRIS-yolk medium, and divided into 3 groups; Group control; Group 2: 10,000 µM of folic acid and in Group 3: 5000 µM of folic acid. Subsequently, the semen samples were packaged in 0.25 mL straws and processed in a semen freezing machine. After a few days, the semen was thawed and evaluated: plasma membrane integrity, acrosome function and integrity. They were analyzed using an analysis of variance (ANOVA) followed by the Newman-Keuls test, using SAS. The addition of a micronutrient with potential antioxidant action in the semen indicates that it can reduce the action of free radicals that alter the plasma membrane and sperm DNA. The evaluation of the plasma membrane integrity, mitochondrial activity and the acrosome integrity of post-thawed sperm were not significantly different between the groups evaluated. Thus, it is concluded that the addition of folic acid in concentrations of 5000 µM and 10000 µM to the TRIS-yolk seminal diluter does not significantly influence the variables evaluated in this experiment.
References
AL-MASKARI, M.Y.; WALY, M.I.; ALI, A.; AL-SHUAIBI, Y.S.; OUHTIT, A. Folate and vitamin B12 deficiency and hyperhomocysteinemia promote oxidative stress in adult type 2 diabetes. Nutrition, v.28, p.23-26, 2012.
CBRA. Colégio Brasileiro de Reprodução Animal, Belo Horizonte, Minas Gerais. Manual para exame andrológico e avaliação de sêmen animal, 2013. 104p.
COLETO, Z.F.; GUERRA, M.M.P.; BATISTA, A.M. Avaliação do sêmen congelado de caprinos com drogas fluorescentes. Revista Brasileira de Medicina Veterinária, São Paulo, v.24, p.101-104, 2002.
FIGUEIREDO, G.C.; REZENDE, M.P.G.; FIGUEIREDO, M.P.; BOZZI, R.; SOUZA JUNIOR, A.A.O.; CARNEIRO, P.L.S.; MALHADO, C.H.M. Morphofunctional characteristics of Dorper sheep crossed with Brazilian native breeds. Small Ruminant Research. v.170, p.143-148, 2019.
GUTHRIE, H.D.; WELCH, G.R. Determination of intracellular reactive oxygen species and high mitochondrial membrane potential in Percoll-treated viable boar sperm using fluorescence-activated flow cytometry. Journal of Animal Science, v. 84, p. 2089-2100, 2006.
JOSHI, R. Free radical scavenging behavior of folic acid: evidence for possible antioxidant activity. Free Radical Biology Medicine, v.30, p.1390-1399, 2001.
MAIA, F.A. Avaliação dos parâmetros seminais de indivíduos inférteis em uso de polivitamínico e polimineral. 2009. 91p. (Dissertação de Mestrado). Faculdade de Medicina de Botucatu, Universidade Estadual Paulista, Botucatu, 2009.
ROTH, T.L.; ANDERSON, G.B.; BONDURANT, R.H.; PASHEN, R.L. Production of sheep X goat hybrid sheep chimeras by inner cell mass transplantation. Biology Reproduction, v.41, p.675-682, 1989.
VIDAL, A.H.; BATISTA, A.M.; SILVA, E.C.B.; GOMES, W.A.; PELINCA, M.A. SILVA, S.V.; GUERRA, M.M. P. Soybean lecithin-based extender as na alternative for goat sperm cryopreservation. Small Ruminant Research, v.109, p.47-51, 2013.
WATSON, P.F. The causes of reduced fertility with cryopreserved. Animal Reproduction Science, v.60–61, p.481–492, 2000.
