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Oviductal Transcriptome Is Modified after Insemination during Spontaneous Ovulation in the Sow.

López-Úbeda R, García-Vázquez FA, Romar R, Gadea J, Muñoz M, Hunter RH, Coy P - PLoS ONE (2015)

Bottom Line: Network 3 integrated the genes with the highest down-regulation level (CYP51, PTH1R and TMOD3).Data in the present study indicate a change in gene expression during gamete encounter at the site of fertilization after a natural sperm selection within the female genital tract.These changes would indicate a modification of the environment preparing the oviduct for a successful fertilization and for an adequate embryo early development.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Veterinary Faculty, University of Murcia, Murcia, Spain; International Excellence Campus for Higher Education and Research (Campus Mare Nostrum), Murcia, Spain.

ABSTRACT
Gene Expression Microarray technology was used to compare oviduct transcriptome between inseminated and non-inseminated pigs during spontaneous oestrus. We used an in vivo model approaching the study from a physiological point of view in which no hormonal treatment (animals were in natural oestrus) and no artificial sperm selection (selection was performed within the female genital) were imposed. It is therefore emphasised that no surgical introduction of spermatozoa and no insemination at a site other than the physiological one were used. This approach revealed 17 genes that were two-fold or more up-regulated in oviducts exposed to spermatozoa and/or developing embryos and 9 genes that were two-fold or more down-regulated. Functional analysis of the genes revealed that the top canonical pathways affected by insemination were related to the inflammatory response and immune system (Network 1) to molecular transport, protein trafficking and developmental disorder (Network 2) and to cell-to-cell signalling and interaction (Network 3). Some of the genes in network 1 had been previously detected in the oviduct of human and animals, where they were over-expressed in the presence of spermatozoa or pre-implantation embryos (C3, IGHG1, ITIH4, TNF and SERPINE1) whereas others were not previously reported (SAA2, ALOX12, CD1D and SPP1). Genes in Network 2 included RAB1B and TOR3A, the latter being described for the first time in the oviduct and clearly expressed in the epithelial cells of the mucosa layer. Network 3 integrated the genes with the highest down-regulation level (CYP51, PTH1R and TMOD3). Data in the present study indicate a change in gene expression during gamete encounter at the site of fertilization after a natural sperm selection within the female genital tract. These changes would indicate a modification of the environment preparing the oviduct for a successful fertilization and for an adequate embryo early development.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemical localization of TOR3A in the porcine oviduct.Strong immunostaining for TOR3A protein was only observed in the oviductal wall of the inseminated sows (Fig 7A, magnified in 7A1). The labelling was absent in non-inseminated animals (Fig 7C magnified in 7C1) as well as in the corresponding negative controls for the inseminated (Fig 7B, magnified in 7B1) and non-inseminated sows (Fig 7D, magnified in 7D1). In all figures, scale bars correspond to 100 μm.
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pone.0130128.g007: Immunohistochemical localization of TOR3A in the porcine oviduct.Strong immunostaining for TOR3A protein was only observed in the oviductal wall of the inseminated sows (Fig 7A, magnified in 7A1). The labelling was absent in non-inseminated animals (Fig 7C magnified in 7C1) as well as in the corresponding negative controls for the inseminated (Fig 7B, magnified in 7B1) and non-inseminated sows (Fig 7D, magnified in 7D1). In all figures, scale bars correspond to 100 μm.

Mentions: Significant gene expression differences derived from microarray analysis were validated by real-time RT-PCR for 8 genes (TOR3A, RAB1B, ALOX12, GSTA1, C3, ITIH4, SPP1 and TMOD3; Table 4, p<0.05). The analysis of the same RNA samples as used for array hybridization allowed a direct comparison of both methods. We further confirmed GSTA1 and TOR3A protein expression in the oviduct by immunohistochemistry (Figs 6 and 7) and the detection of TOR3A by Western blotting into the oviductal fluid (Fig 8).


Oviductal Transcriptome Is Modified after Insemination during Spontaneous Ovulation in the Sow.

López-Úbeda R, García-Vázquez FA, Romar R, Gadea J, Muñoz M, Hunter RH, Coy P - PLoS ONE (2015)

Immunohistochemical localization of TOR3A in the porcine oviduct.Strong immunostaining for TOR3A protein was only observed in the oviductal wall of the inseminated sows (Fig 7A, magnified in 7A1). The labelling was absent in non-inseminated animals (Fig 7C magnified in 7C1) as well as in the corresponding negative controls for the inseminated (Fig 7B, magnified in 7B1) and non-inseminated sows (Fig 7D, magnified in 7D1). In all figures, scale bars correspond to 100 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476686&req=5

pone.0130128.g007: Immunohistochemical localization of TOR3A in the porcine oviduct.Strong immunostaining for TOR3A protein was only observed in the oviductal wall of the inseminated sows (Fig 7A, magnified in 7A1). The labelling was absent in non-inseminated animals (Fig 7C magnified in 7C1) as well as in the corresponding negative controls for the inseminated (Fig 7B, magnified in 7B1) and non-inseminated sows (Fig 7D, magnified in 7D1). In all figures, scale bars correspond to 100 μm.
Mentions: Significant gene expression differences derived from microarray analysis were validated by real-time RT-PCR for 8 genes (TOR3A, RAB1B, ALOX12, GSTA1, C3, ITIH4, SPP1 and TMOD3; Table 4, p<0.05). The analysis of the same RNA samples as used for array hybridization allowed a direct comparison of both methods. We further confirmed GSTA1 and TOR3A protein expression in the oviduct by immunohistochemistry (Figs 6 and 7) and the detection of TOR3A by Western blotting into the oviductal fluid (Fig 8).

Bottom Line: Network 3 integrated the genes with the highest down-regulation level (CYP51, PTH1R and TMOD3).Data in the present study indicate a change in gene expression during gamete encounter at the site of fertilization after a natural sperm selection within the female genital tract.These changes would indicate a modification of the environment preparing the oviduct for a successful fertilization and for an adequate embryo early development.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Veterinary Faculty, University of Murcia, Murcia, Spain; International Excellence Campus for Higher Education and Research (Campus Mare Nostrum), Murcia, Spain.

ABSTRACT
Gene Expression Microarray technology was used to compare oviduct transcriptome between inseminated and non-inseminated pigs during spontaneous oestrus. We used an in vivo model approaching the study from a physiological point of view in which no hormonal treatment (animals were in natural oestrus) and no artificial sperm selection (selection was performed within the female genital) were imposed. It is therefore emphasised that no surgical introduction of spermatozoa and no insemination at a site other than the physiological one were used. This approach revealed 17 genes that were two-fold or more up-regulated in oviducts exposed to spermatozoa and/or developing embryos and 9 genes that were two-fold or more down-regulated. Functional analysis of the genes revealed that the top canonical pathways affected by insemination were related to the inflammatory response and immune system (Network 1) to molecular transport, protein trafficking and developmental disorder (Network 2) and to cell-to-cell signalling and interaction (Network 3). Some of the genes in network 1 had been previously detected in the oviduct of human and animals, where they were over-expressed in the presence of spermatozoa or pre-implantation embryos (C3, IGHG1, ITIH4, TNF and SERPINE1) whereas others were not previously reported (SAA2, ALOX12, CD1D and SPP1). Genes in Network 2 included RAB1B and TOR3A, the latter being described for the first time in the oviduct and clearly expressed in the epithelial cells of the mucosa layer. Network 3 integrated the genes with the highest down-regulation level (CYP51, PTH1R and TMOD3). Data in the present study indicate a change in gene expression during gamete encounter at the site of fertilization after a natural sperm selection within the female genital tract. These changes would indicate a modification of the environment preparing the oviduct for a successful fertilization and for an adequate embryo early development.

No MeSH data available.


Related in: MedlinePlus