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...
Published in: | PeerJ |
---|---|
Main Authors: | , , , , , , , |
Other Authors: | |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
PeerJ
2017
|
Subjects: | |
Online Access: | http://dx.doi.org/10.7717/peerj.3840 https://peerj.com/articles/3840.pdf https://peerj.com/articles/3840.xml https://peerj.com/articles/3840.html |
id |
crpeerj:10.7717/peerj.3840 |
---|---|
record_format |
openpolar |
spelling |
crpeerj:10.7717/peerj.3840 2024-06-02T08:04:16+00:00 A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterus leucas) Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng Ministry of Science and Technology 2017 http://dx.doi.org/10.7717/peerj.3840 https://peerj.com/articles/3840.pdf https://peerj.com/articles/3840.xml https://peerj.com/articles/3840.html en eng PeerJ http://creativecommons.org/licenses/by/4.0/ PeerJ volume 5, page e3840 ISSN 2167-8359 journal-article 2017 crpeerj https://doi.org/10.7717/peerj.3840 2024-05-07T14:14:20Z 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 RNA later ™ 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 PeerJ Publishing PeerJ 5 e3840 |
institution |
Open Polar |
collection |
PeerJ Publishing |
op_collection_id |
crpeerj |
language |
English |
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 RNA later ™ 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. |
author2 |
Ministry of Science and Technology |
format |
Article in Journal/Newspaper |
author |
Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng |
spellingShingle |
Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterus leucas) |
author_facet |
Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng |
author_sort |
Tsai, Ming-An |
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 |
publishDate |
2017 |
url |
http://dx.doi.org/10.7717/peerj.3840 https://peerj.com/articles/3840.pdf https://peerj.com/articles/3840.xml https://peerj.com/articles/3840.html |
genre |
Beluga Beluga* Delphinapterus leucas |
genre_facet |
Beluga Beluga* Delphinapterus leucas |
op_source |
PeerJ volume 5, page e3840 ISSN 2167-8359 |
op_rights |
http://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.7717/peerj.3840 |
container_title |
PeerJ |
container_volume |
5 |
container_start_page |
e3840 |
_version_ |
1800748899213770752 |