Characterization of the Impact of Density Gradient Centrifugation on the Profile of the Pig Sperm Transcriptome by RNA-Seq
Lian, Yu (Centre de Recerca en Agrigenòmica)
Gòdia, Marta (Centre de Recerca en Agrigenòmica)
Castelló Farré, Anna (Universitat Autònoma de Barcelona. Departament de Ciència Animal i dels Aliments)
Rodríguez Gil, Joan Enric (Universitat Autònoma de Barcelona. Departament de Medicina i Cirurgia Animals)
Balasch, Sam (Grup Gepork S.A. (Barcelona))
Sánchez Bonastre, Armando (Centre de Recerca en Agrigenòmica)
Clop, Alex (Centre de Recerca en Agrigenòmica)

Data:	2021
Resum:	RNA-Seq data from human semen suggests that the study of the sperm transcriptome requires the previous elimination from the ejaculates of somatic cells carrying a larger load of RNA. Semen purification is also carried to study the sperm transcriptome in other species including swine and it is often done by density gradient centrifugation to obtain viable spermatozoa from fresh ejaculates or artificial insemination doses, thereby limiting the throughput and remoteness of the samples that can be processed in one study. The aim of this work was to evaluate the impact of purification with density gradient centrifugation by BoviPure TM on porcine sperm. Four boar ejaculates were purified with BoviPure TM and their transcriptome sequenced by RNA-Seq was compared with the RNA-Seq profiles of their paired non-purified sample. Seven thousand five hundred and nineteen protein coding genes were identified. Correlation, cluster, and principal component analysis indicated high-although not complete-similarity between the purified and the paired non-purified ejaculates. 372 genes displayed differentially abundant RNA levels between treatments. Most of these genes had lower abundances after purification and were mostly related to translation, transcription and metabolic processes. We detected a significant change in the proportion of genes of epididymal origin within the differentially abundant genes (1. 3%) when compared with the catalog of unaltered genes (0. 2%). In contrast, the proportion of testis-specific genes was higher in the group of unaltered genes (4%) when compared to the list of differentially abundant genes (0%). No proportion differences were identified for prostate, white blood, lymph node, tonsil, duodenum, skeletal muscle, liver, and mammary gland. Altogether, these results suggest that the purification impacts on the RNA levels of a small number of genes which are most likely caused by the removal of epididymal epithelial cells but also premature germinal cells, immature or abnormal spermatozoa or seminal exosomes with a distinct load of RNAs.
Ajuts:	Ministerio de Economía y Competitividad AGL2013-44978-R Agencia Estatal de Investigación AGL2017-86946-R Agència de Gestió d'Ajuts Universitaris i de Recerca 2014/SGR-1528 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-1060 Ministerio de Economía y Competitividad SEV-2015-0533 Agencia Estatal de Investigación CEX2019-000902-S
Nota:	Altres ajuts: CERCA Programme/Generalitat de Catalunya
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Sperm RNA ; RNA-Seq ; Sperm purification ; Differentially abundant gene ; Somatic cell ; Germline cell ; Exosome
Publicat a:	Frontiers in Veterinary Science, Vol. 8 (July 2021) , art. 668158, ISSN 2297-1769

DOI: 10.3389/fvets.2021.668158
PMID: 34350225

12 p, 714.2 KB

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