Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs
Alternative splicing is a cellular mechanism in eukaryotes that results in considerable diversity of gene products. It plays an important role in several diseases and cellular signal regulation. Heat stress is a major factor that induces immunosuppression in pigs. Little is known about the correlati...
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KeAi Communications Co., Ltd.
2014-11-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311913605742 |
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| author | Xiang-hong JU Han-jin XU Yan-hong YONG Li-long AN Ying-mei XU Pei-rong JIAO Ming LIAO |
| author_facet | Xiang-hong JU Han-jin XU Yan-hong YONG Li-long AN Ying-mei XU Pei-rong JIAO Ming LIAO |
| author_sort | Xiang-hong JU |
| collection | DOAJ |
| description | Alternative splicing is a cellular mechanism in eukaryotes that results in considerable diversity of gene products. It plays an important role in several diseases and cellular signal regulation. Heat stress is a major factor that induces immunosuppression in pigs. Little is known about the correlation between alternative splicing and heat stress in pigs. Therefore, this study aimed to clone, sequence and quantify the alternative splicing variant of toll-like receptor 4 (TLR4) in Bama miniature pigs (Sus scrofa domestica) following exposure to heat stress. The results showed that the second exon of TLR4 was spliced and 167 bp shorter in the alternative splicing variant, and the protein was putatively identified as a type of truncated membrane protein consisting of extramembrane, transmembrane and intramembrane regions lacking a signal peptide. Further, it was not a nonclassical secretory protein. Five potential reference genes were screened for their potential as reliable standards to quantify the expression of TLR4 alternative spliced variants by real-time quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). The stability of these reference genes was ranked using the geNorm and NormFinder programs, and ribosomal protein L4 (RPL4) and TATA box-binding protein (TBP) were found to be the two genes showing the most stable expression in the in vitro cultured peripheral blood mononuclear cells (PBMCs) during heat shock. The mRNA level of the TLR4 gene (both classical and spliced) in stressed pigs increased significantly (P<0.05). Further, the expression levels of the alternative spliced variant of TLR4 (TLR4-ASV) showed a 2–3 folds increase in heat-stressed PBMCs as compared to control pigs. The results of the present study suggested that heat shock might modulate the host immune response by regulating the expressions of TLR4 and its alternative splicing variant. |
| format | Article |
| id | doaj-art-3c3e5246bed0467bb7095a05a686ef6a |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2014-11-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-3c3e5246bed0467bb7095a05a686ef6a2025-08-20T03:58:23ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192014-11-0113112479248710.1016/S2095-3119(13)60574-2Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature PigsXiang-hong JU0Han-jin XU1Yan-hong YONG2Li-long AN3Ying-mei XU4Pei-rong JIAO5Ming LIAO6Department of Veterinary Medicine, Guangdong Ocean University, Zhanjiang 524088, P.R. China; Key Laboratory of Animal Disease Control and Prevention, Ministry of Agriculture/College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R. China; JU Xiang-hong, Mobile: 13553523590Department of Animal Science, Guangdong Ocean University, Zhanjiang 524088, P.R. ChinaDepartment of Veterinary Medicine, Guangdong Ocean University, Zhanjiang 524088, P.R. ChinaDepartment of Animal Science, Guangdong Ocean University, Zhanjiang 524088, P.R. ChinaDepartment of Animal Science, Guangdong Ocean University, Zhanjiang 524088, P.R. ChinaKey Laboratory of Animal Disease Control and Prevention, Ministry of Agriculture/College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R. ChinaKey Laboratory of Animal Disease Control and Prevention, Ministry of Agriculture/College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R. China; Correspondence LIAO Ming, Tel/Fax: +86-20-85280240Alternative splicing is a cellular mechanism in eukaryotes that results in considerable diversity of gene products. It plays an important role in several diseases and cellular signal regulation. Heat stress is a major factor that induces immunosuppression in pigs. Little is known about the correlation between alternative splicing and heat stress in pigs. Therefore, this study aimed to clone, sequence and quantify the alternative splicing variant of toll-like receptor 4 (TLR4) in Bama miniature pigs (Sus scrofa domestica) following exposure to heat stress. The results showed that the second exon of TLR4 was spliced and 167 bp shorter in the alternative splicing variant, and the protein was putatively identified as a type of truncated membrane protein consisting of extramembrane, transmembrane and intramembrane regions lacking a signal peptide. Further, it was not a nonclassical secretory protein. Five potential reference genes were screened for their potential as reliable standards to quantify the expression of TLR4 alternative spliced variants by real-time quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). The stability of these reference genes was ranked using the geNorm and NormFinder programs, and ribosomal protein L4 (RPL4) and TATA box-binding protein (TBP) were found to be the two genes showing the most stable expression in the in vitro cultured peripheral blood mononuclear cells (PBMCs) during heat shock. The mRNA level of the TLR4 gene (both classical and spliced) in stressed pigs increased significantly (P<0.05). Further, the expression levels of the alternative spliced variant of TLR4 (TLR4-ASV) showed a 2–3 folds increase in heat-stressed PBMCs as compared to control pigs. The results of the present study suggested that heat shock might modulate the host immune response by regulating the expressions of TLR4 and its alternative splicing variant.http://www.sciencedirect.com/science/article/pii/S2095311913605742alternative splicing variantBama miniature pigtoll-like receptor 4heat stress |
| spellingShingle | Xiang-hong JU Han-jin XU Yan-hong YONG Li-long AN Ying-mei XU Pei-rong JIAO Ming LIAO Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs Journal of Integrative Agriculture alternative splicing variant Bama miniature pig toll-like receptor 4 heat stress |
| title | Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs |
| title_full | Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs |
| title_fullStr | Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs |
| title_full_unstemmed | Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs |
| title_short | Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs |
| title_sort | heat stress upregulates the expression of tlr4 and its alternative splicing variant in bama miniature pigs |
| topic | alternative splicing variant Bama miniature pig toll-like receptor 4 heat stress |
| url | http://www.sciencedirect.com/science/article/pii/S2095311913605742 |
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