Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae

Despite the reported promising farrowing rates after non-surgical and surgical transfers of vitrified porcine morulae and blastocysts produced in vivo (range: 70–75%), the pregnancy loss is 5–15 fold higher with vitrified than with fresh embryos. The present study aimed to investigate whether vitrif...

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Autores principales:	Cristina Cuello, Cristina A. Martinez, Josep M. Cambra, Alejandro González-Plaza, Inmaculada Parrilla, Heriberto Rodriguez-Martinez, Maria A. Gil, Emilio A. Martinez
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	morula vitrification transcriptome gene expression pig embryo Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/5c716cc938be410892888ee6fe14ad77
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spelling	oai:doaj.org-article:5c716cc938be410892888ee6fe14ad772021-11-12T05:49:43ZVitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae2297-176910.3389/fvets.2021.771996https://doaj.org/article/5c716cc938be410892888ee6fe14ad772021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fvets.2021.771996/fullhttps://doaj.org/toc/2297-1769Despite the reported promising farrowing rates after non-surgical and surgical transfers of vitrified porcine morulae and blastocysts produced in vivo (range: 70–75%), the pregnancy loss is 5–15 fold higher with vitrified than with fresh embryos. The present study aimed to investigate whether vitrification affects the transcriptome of porcine morulae, using microarrays and RT-qPCR validation. Morulae were obtained surgically from weaned sows (n = 13) on day 6 (day 0 = estrus onset). A total of 60 morulae were vitrified (treatment group). After 1 week of storage, the vitrified morulae were warmed. Vitrified-warmed and non-vitrified fresh morulae (control; n = 40) were cultured for 24 h to assess embryo survival by stereomicroscopy after. A total of 30 vitrified/warmed embryos that were deemed viable and 30 fresh control embryos (three pools of 10 for each experimental group) were selected for microarray analysis. Gene expression was assessed with a GeneChip® Porcine Genome Array (Affymetrix). An ANOVA analysis p-unadjusted <0.05 and a fold change cut-off of ±1.5 were set to identify differentially expressed genes (DEGs). Data analysis and biological interpretation were performed using the Partek Genomic Suite 7.0 software. The survival rate of morulae after vitrification and warming (92.0 ± 8.3%) was similar to that of the control (100%). A total of 233 DEGs were identified in vitrified morulae (38 upregulated and 195 downregulated), compared to the control group. Nine pathways were significantly modified. Go-enrichment analysis revealed that DEGs were mainly related to the Biological Process functional group. Up-regulated DEGs were involved in glycosaminoglycan degradation, metabolic pathways and tryptophan metabolism KEGG pathways. The pathways related to the down-regulated DEGs were glycolysis/gluconeogenesis, protein export and fatty acid elongation. The disruption of metabolic pathways in morulae could be related to impaired embryo quality and developmental potential, despite the relatively high survival rates after warming observed in vitro. In conclusion, vitrification altered the gene expression pattern of porcine morulae produced in vivo, generating alterations in the transcriptome that may interfere with subsequent embryo development and pregnancy after embryo transfer.Cristina CuelloCristina A. MartinezJosep M. CambraAlejandro González-PlazaInmaculada ParrillaHeriberto Rodriguez-MartinezMaria A. GilEmilio A. MartinezFrontiers Media S.A.articlemorulavitrificationtranscriptomegene expressionpigembryoVeterinary medicineSF600-1100ENFrontiers in Veterinary Science, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	morula vitrification transcriptome gene expression pig embryo Veterinary medicine SF600-1100
spellingShingle	morula vitrification transcriptome gene expression pig embryo Veterinary medicine SF600-1100 Cristina Cuello Cristina A. Martinez Josep M. Cambra Alejandro González-Plaza Inmaculada Parrilla Heriberto Rodriguez-Martinez Maria A. Gil Emilio A. Martinez Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae
description	Despite the reported promising farrowing rates after non-surgical and surgical transfers of vitrified porcine morulae and blastocysts produced in vivo (range: 70–75%), the pregnancy loss is 5–15 fold higher with vitrified than with fresh embryos. The present study aimed to investigate whether vitrification affects the transcriptome of porcine morulae, using microarrays and RT-qPCR validation. Morulae were obtained surgically from weaned sows (n = 13) on day 6 (day 0 = estrus onset). A total of 60 morulae were vitrified (treatment group). After 1 week of storage, the vitrified morulae were warmed. Vitrified-warmed and non-vitrified fresh morulae (control; n = 40) were cultured for 24 h to assess embryo survival by stereomicroscopy after. A total of 30 vitrified/warmed embryos that were deemed viable and 30 fresh control embryos (three pools of 10 for each experimental group) were selected for microarray analysis. Gene expression was assessed with a GeneChip® Porcine Genome Array (Affymetrix). An ANOVA analysis p-unadjusted <0.05 and a fold change cut-off of ±1.5 were set to identify differentially expressed genes (DEGs). Data analysis and biological interpretation were performed using the Partek Genomic Suite 7.0 software. The survival rate of morulae after vitrification and warming (92.0 ± 8.3%) was similar to that of the control (100%). A total of 233 DEGs were identified in vitrified morulae (38 upregulated and 195 downregulated), compared to the control group. Nine pathways were significantly modified. Go-enrichment analysis revealed that DEGs were mainly related to the Biological Process functional group. Up-regulated DEGs were involved in glycosaminoglycan degradation, metabolic pathways and tryptophan metabolism KEGG pathways. The pathways related to the down-regulated DEGs were glycolysis/gluconeogenesis, protein export and fatty acid elongation. The disruption of metabolic pathways in morulae could be related to impaired embryo quality and developmental potential, despite the relatively high survival rates after warming observed in vitro. In conclusion, vitrification altered the gene expression pattern of porcine morulae produced in vivo, generating alterations in the transcriptome that may interfere with subsequent embryo development and pregnancy after embryo transfer.
format	article
author	Cristina Cuello Cristina A. Martinez Josep M. Cambra Alejandro González-Plaza Inmaculada Parrilla Heriberto Rodriguez-Martinez Maria A. Gil Emilio A. Martinez
author_facet	Cristina Cuello Cristina A. Martinez Josep M. Cambra Alejandro González-Plaza Inmaculada Parrilla Heriberto Rodriguez-Martinez Maria A. Gil Emilio A. Martinez
author_sort	Cristina Cuello
title	Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae
title_short	Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae
title_full	Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae
title_fullStr	Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae
title_full_unstemmed	Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae
title_sort	vitrification effects on the transcriptome of in vivo-derived porcine morulae
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/5c716cc938be410892888ee6fe14ad77
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Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae

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