MicroRNA profile analysis in human and mice sperm collected from different regions of the reproductive tract

Abstract

World-wide 50% of human infertility is due to male infertility problems. Thus, assisted reproductive technologies (ARTs), such as testicular sperm extraction and aspiration, are employed to overcome these infertility problems. However, recent reports indicated that the employment of ARTs may be associated with an increased in the incidence of congenital malformations and epigenetic defects on the next generations. Sperm non-coding RNAs (ncRNAs), such as microRNAs (miRNA) and tsRNAs, are non-coding RNAs molecules involved in controlling gene levels by posttranscription and transcription mechanisms, thus playing an important role in developmental biology. Besides, the ncRNAs can escape the reprograming process, which means that they can be transmitted to the next generation characterizing an intergenerational transfer. Specific sperm borne miRNAs can induced epigenetic changes, when injected in the zygote, thus leading to alterations present in the sperm donor. Thus, our hypothesis is that sperm obtained from testis of men and mice (immature sperm) present a different miRNAs expression compared to sperm obtained from epididymis cauda (mature sperm) and from sperm ejaculated (in human). For this, we will obtain and purify sperm from testis and epididymis cauda from men and mice and ejaculated from men. Total RNA extraction will be performed and microarray analyzes will be performed. Then, data will be analyzed by bioinformatics analyses to verify relation with embryo development. Analysis of variance will be performed to verify differences between miRNA expressions. Tukey test will be employed when necessary. The present study will elucidated the difference between miRNA profiles of sperm and will indicate if there is a relation with embryo development and epigenetic alterations. (AU)