Capillary electroporation affects the expression of miRNA-122-5p from bull sperm cells.

Gene 2021 Feb 05

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Sperm-mediated gene transfer (SMGT) has a potential application in the generation of transgenic animals. Capillary electroporation consists of the application of electrical pulses, resulting in an increased transfection rate. Little is known about the impacts of the transfection of exogenous DNA on sperm epigenetics. MicroRNAs are epigenetic factors that are related to sperm motility. MiRNA-122-5p regulates genes that influence motility, and consequently, the fertilizing potential of sperm. Therefore, we aimed at identifying whether epigenetic factors such as microRNAs could be altered after DNA transfection, using the capillary electroporation technique. In this study, bull sperm was electroporated using voltages of 600 V, 1500 V, and 0 V (control group), with or without exogenous DNA. Parameters of sperm quality were analyzed using CASA and flow cytometry, and expression of the miRNA-122-5p was analyzed using RT-qPCR. It was observed that electroporation increased the internalization of exogenous DNA (P < 0.05), but did not impair the mitochondrial activity (P > 0.05). It reduced sperm motility (P < 0.05). The expression of miRNA-122-5p was upregulated in sperm electroporated at 1500 V, and the presence of exogenous DNA did not affect its expression. Thus, we can conclude that electroporation influences the expression of miRNA-122-5p from bull sperm cells. Copyright © 2020 Elsevier B.V. All rights reserved.

Citation

Lucas Dos Santos da Silva, William Borges Domingues, Bruna Fagundes Barreto, Amanda Weege da Silveira Martins, Eduardo Nunes Dellagostin, Eliza Rossi Komninou, Carine D Corcini, Antônio Sergio Varela Junior, Vinicius Farias Campos. Capillary electroporation affects the expression of miRNA-122-5p from bull sperm cells. Gene. 2021 Feb 05;768:145286