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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ftdoajarticles:oai:doaj.org/article:87ca00172a0e453e84636fe79f9cc691 2024-01-07T09:42:26+01:00 A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas) Ming-An Tsai I-Hua Chen Jiann-Hsiung Wang Shih-Jen Chou Tsung-Hsien Li Ming-Yih Leu Hsiao-Kuan Ho Wei Cheng Yang 2017-09-01T00:00:00Z https://doi.org/10.7717/peerj.3840 https://doaj.org/article/87ca00172a0e453e84636fe79f9cc691 EN eng PeerJ Inc. https://peerj.com/articles/3840.pdf https://peerj.com/articles/3840/ https://doaj.org/toc/2167-8359 doi:10.7717/peerj.3840 2167-8359 https://doaj.org/article/87ca00172a0e453e84636fe79f9cc691 PeerJ, Vol 5, p e3840 (2017) Gene expression qRT-PCR Immunology Beluga Medicine R Biology (General) QH301-705.5 article 2017 ftdoajarticles https://doi.org/10.7717/peerj.3840 2023-12-10T01:51:13Z 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. Article in Journal/Newspaper Beluga Beluga* Delphinapterus leucas Directory of Open Access Journals: DOAJ Articles PeerJ 5 e3840 |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Gene expression qRT-PCR Immunology Beluga Medicine R Biology (General) QH301-705.5 |
spellingShingle |
Gene expression qRT-PCR Immunology Beluga Medicine R Biology (General) QH301-705.5 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) |
topic_facet |
Gene expression qRT-PCR Immunology Beluga Medicine R Biology (General) QH301-705.5 |
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. |
format |
Article in Journal/Newspaper |
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 |
title |
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_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_sort |
probe-based qrt-pcr method to profile immunological gene expression in blood of captive beluga whales (delphinapterus leucas) |
publisher |
PeerJ Inc. |
publishDate |
2017 |
url |
https://doi.org/10.7717/peerj.3840 https://doaj.org/article/87ca00172a0e453e84636fe79f9cc691 |
genre |
Beluga Beluga* Delphinapterus leucas |
genre_facet |
Beluga Beluga* Delphinapterus leucas |
op_source |
PeerJ, Vol 5, p e3840 (2017) |
op_relation |
https://peerj.com/articles/3840.pdf https://peerj.com/articles/3840/ https://doaj.org/toc/2167-8359 doi:10.7717/peerj.3840 2167-8359 https://doaj.org/article/87ca00172a0e453e84636fe79f9cc691 |
op_doi |
https://doi.org/10.7717/peerj.3840 |
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PeerJ |
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5 |
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e3840 |
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1787423407909896192 |