A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas)
Cytokines are fundamental for a functioning immune system, and thus potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for...
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PeerJ Inc.
2017-09-01
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| Online Access: | https://peerj.com/articles/3840.pdf |
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| author | Ming-An Tsai I-Hua Chen Jiann-Hsiung Wang Shih-Jen Chou Tsung-Hsien Li Ming-Yih Leu Hsiao-Kuan Ho Wei Cheng Yang |
| author_facet | Ming-An Tsai I-Hua Chen Jiann-Hsiung Wang Shih-Jen Chou Tsung-Hsien Li Ming-Yih Leu Hsiao-Kuan Ho Wei Cheng Yang |
| author_sort | Ming-An Tsai |
| collection | DOAJ |
| description | Cytokines are fundamental for a functioning immune system, and thus potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a probe-based quantitative gene expression assay for immunological assessment of captive beluga whales (Delphinapterus leucas) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater™ solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset. |
| first_indexed | 2024-03-09T06:59:15Z |
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| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T06:59:15Z |
| publishDate | 2017-09-01 |
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| series | PeerJ |
| spelling | doaj.art-87ca00172a0e453e84636fe79f9cc6912023-12-03T09:57:00ZengPeerJ Inc.PeerJ2167-83592017-09-015e384010.7717/peerj.3840A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas)Ming-An Tsai0I-Hua Chen1Jiann-Hsiung Wang2Shih-Jen Chou3Tsung-Hsien Li4Ming-Yih Leu5Hsiao-Kuan Ho6Wei Cheng Yang7Department of Biology, National Museum of Marine Biology and Aquarium, Pingtung, TaiwanCollege of Veterinary Medicine, National Chiayi University, Chiayi, TaiwanCollege of Veterinary Medicine, National Chiayi University, Chiayi, TaiwanCollege of Veterinary Medicine, National Chiayi University, Chiayi, TaiwanDepartment of Biology, National Museum of Marine Biology and Aquarium, Pingtung, TaiwanDepartment of Biology, National Museum of Marine Biology and Aquarium, Pingtung, TaiwanDepartment of Biology, Hi-Scene World Enterprise Co. Ltd., Pingtung, TaiwanCollege of Veterinary Medicine, National Chiayi University, Chiayi, TaiwanCytokines are fundamental for a functioning immune system, and thus potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a probe-based quantitative gene expression assay for immunological assessment of captive beluga whales (Delphinapterus leucas) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater™ solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset.https://peerj.com/articles/3840.pdfGene expressionqRT-PCRImmunologyBeluga |
| spellingShingle | Ming-An Tsai I-Hua Chen Jiann-Hsiung Wang Shih-Jen Chou Tsung-Hsien Li Ming-Yih Leu Hsiao-Kuan Ho Wei Cheng Yang A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas) PeerJ Gene expression qRT-PCR Immunology Beluga |
| title | A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas) |
| title_full | A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas) |
| title_fullStr | A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas) |
| title_full_unstemmed | A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas) |
| title_short | A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas) |
| title_sort | probe based qrt pcr method to profile immunological gene expression in blood of captive beluga whales delphinapterus leucas |
| topic | Gene expression qRT-PCR Immunology Beluga |
| url | https://peerj.com/articles/3840.pdf |
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