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Detection of differentially expressed genes between Erhualian and Large White placentas on day 75 and 90 of gestation.

Zhou QY, Fang MD, Huang TH, Li CC, Yu M, Zhao SH - BMC Genomics (2009)

Bottom Line: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality.Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds.Differential expression of six candidate imprinted genes was also confirmed.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, PR China. zhouquanyong@webmail.hzau.edu.cn

ABSTRACT

Background: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality. Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds. To understand the molecular basis of placental development during late gestation in Chinese indigenous and Western breeds with different placental efficiency, female placental samples were collected from six pregnant Erhualian gilts at gestation day 75 (E75) and day 90 (E90) and from six pregnant Large White gilts at gestation day 75 (L75) and day 90 (L90). Two female placentas from one sow were used to extract RNA and then pooled in equal volumes. Twelve pooled samples were hybridized to the porcine Affymetrix GeneChip.

Results: A total of 226 and 577 transcripts were detected that were differentially expressed between E75 and L75 and between E90 and L90 (p < 0.01, q < 0.2), respectively. Gene Ontology (GO) analysis revealed that these genes belong to the class of genes that participate in angiogenesis and development. Real-time RT-PCR confirmed the differential expression of eight selected genes. Significant differential expression of five genes in the VEGF pathway was also detected between the breeds. A search of chromosomal location revealed that 44 differentially expressed genes located to QTL regions related to reproduction. Differential expression of six candidate imprinted genes was also confirmed. Three of the six genes (PLAGL1, DIRAS3, and SLC38A4) showed monoallelic expression in the porcine placenta.

Conclusion: Our study detected many genes that showed differential expression between placentas of two divergent breed of pigs, and confirmed the imprinting of three genes. These findings help to elucidate the genetic control of placental efficiency and improve the understanding of placental development.

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Real time RT-PCR results of 13 genes. (A) Validation of the Microarray data by real-time RT-PCR analysis of eight representative genes. Expression levels of eight genes were detected in E75, L75, E90 and L90 by real-time RT-PCR (red) and microarray (blue). R represents the Pearson correlation coefficient. (B) real-time RT-PCR results of genes related to vascular development and vascular permeability. The x-axis represents the genes and the y-axis shows the fold change in expression. Different superscript alphanumeric characters indicate a statistically significant difference at p < 0.05.
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Figure 2: Real time RT-PCR results of 13 genes. (A) Validation of the Microarray data by real-time RT-PCR analysis of eight representative genes. Expression levels of eight genes were detected in E75, L75, E90 and L90 by real-time RT-PCR (red) and microarray (blue). R represents the Pearson correlation coefficient. (B) real-time RT-PCR results of genes related to vascular development and vascular permeability. The x-axis represents the genes and the y-axis shows the fold change in expression. Different superscript alphanumeric characters indicate a statistically significant difference at p < 0.05.

Mentions: Thirteen genes (ALDH1A1, DIO3, DIRAS3, PLAGL1, PON2, ASCL2, WIF1, SLC38A4, VEGF, VEGFR-1, VEGFR-2, VE-cadherin and β-arrestin2) were selected to confirm the expression by the use of real-time RT-PCR. Among these genes, eight genes were used to validate the microarray data. The results indicated that expression patterns of these genes were consistent with the microarray (Pearson correlation coefficient >0.75, Figure 2A). The other five genes play important roles in placental angiogenesis and vascular permeability, so we also detected their expression by real-time RT-PCR. Four genes showed more than two-fold differences between breeds in the microarray analysis, but were not statistically significant. Gene β-arrestin2 is another important gene in the VEGF pathway but is not on the microarray. Expression of VEGFR-1 was significantly lower in E75 than L75, but significantly higher in E90 than L90. VEGFR-2 showed significantly higher expression in E90 than in L90. The expression levels of the VE-cadherin and β-arrestin2 genes were significantly lower in L90 than L75.


Detection of differentially expressed genes between Erhualian and Large White placentas on day 75 and 90 of gestation.

Zhou QY, Fang MD, Huang TH, Li CC, Yu M, Zhao SH - BMC Genomics (2009)

Real time RT-PCR results of 13 genes. (A) Validation of the Microarray data by real-time RT-PCR analysis of eight representative genes. Expression levels of eight genes were detected in E75, L75, E90 and L90 by real-time RT-PCR (red) and microarray (blue). R represents the Pearson correlation coefficient. (B) real-time RT-PCR results of genes related to vascular development and vascular permeability. The x-axis represents the genes and the y-axis shows the fold change in expression. Different superscript alphanumeric characters indicate a statistically significant difference at p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2724418&req=5

Figure 2: Real time RT-PCR results of 13 genes. (A) Validation of the Microarray data by real-time RT-PCR analysis of eight representative genes. Expression levels of eight genes were detected in E75, L75, E90 and L90 by real-time RT-PCR (red) and microarray (blue). R represents the Pearson correlation coefficient. (B) real-time RT-PCR results of genes related to vascular development and vascular permeability. The x-axis represents the genes and the y-axis shows the fold change in expression. Different superscript alphanumeric characters indicate a statistically significant difference at p < 0.05.
Mentions: Thirteen genes (ALDH1A1, DIO3, DIRAS3, PLAGL1, PON2, ASCL2, WIF1, SLC38A4, VEGF, VEGFR-1, VEGFR-2, VE-cadherin and β-arrestin2) were selected to confirm the expression by the use of real-time RT-PCR. Among these genes, eight genes were used to validate the microarray data. The results indicated that expression patterns of these genes were consistent with the microarray (Pearson correlation coefficient >0.75, Figure 2A). The other five genes play important roles in placental angiogenesis and vascular permeability, so we also detected their expression by real-time RT-PCR. Four genes showed more than two-fold differences between breeds in the microarray analysis, but were not statistically significant. Gene β-arrestin2 is another important gene in the VEGF pathway but is not on the microarray. Expression of VEGFR-1 was significantly lower in E75 than L75, but significantly higher in E90 than L90. VEGFR-2 showed significantly higher expression in E90 than in L90. The expression levels of the VE-cadherin and β-arrestin2 genes were significantly lower in L90 than L75.

Bottom Line: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality.Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds.Differential expression of six candidate imprinted genes was also confirmed.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, PR China. zhouquanyong@webmail.hzau.edu.cn

ABSTRACT

Background: Placental efficiency is strongly associated with litter size, fetal weight and prenatal mortality. Together with its rapid growth during late gestation, the Large White pig breed shows a significant increase in placental size and weight, but this does not occur in the highly prolific Chinese pig breeds. To understand the molecular basis of placental development during late gestation in Chinese indigenous and Western breeds with different placental efficiency, female placental samples were collected from six pregnant Erhualian gilts at gestation day 75 (E75) and day 90 (E90) and from six pregnant Large White gilts at gestation day 75 (L75) and day 90 (L90). Two female placentas from one sow were used to extract RNA and then pooled in equal volumes. Twelve pooled samples were hybridized to the porcine Affymetrix GeneChip.

Results: A total of 226 and 577 transcripts were detected that were differentially expressed between E75 and L75 and between E90 and L90 (p < 0.01, q < 0.2), respectively. Gene Ontology (GO) analysis revealed that these genes belong to the class of genes that participate in angiogenesis and development. Real-time RT-PCR confirmed the differential expression of eight selected genes. Significant differential expression of five genes in the VEGF pathway was also detected between the breeds. A search of chromosomal location revealed that 44 differentially expressed genes located to QTL regions related to reproduction. Differential expression of six candidate imprinted genes was also confirmed. Three of the six genes (PLAGL1, DIRAS3, and SLC38A4) showed monoallelic expression in the porcine placenta.

Conclusion: Our study detected many genes that showed differential expression between placentas of two divergent breed of pigs, and confirmed the imprinting of three genes. These findings help to elucidate the genetic control of placental efficiency and improve the understanding of placental development.

Show MeSH