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Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells
BACKGROUND: A comprehensive understanding of the equine circadian clock involves the evaluation of circadian clock gene expression. A non-invasive and effective method for detecting equine clock gene expression has yet to be established. Currently, research surrounding this area has relied on collec...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3514281/ https://www.ncbi.nlm.nih.gov/pubmed/23039139 http://dx.doi.org/10.1186/1740-3391-10-7 |
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author | Watts, Lisa M Browne, John A Murphy, Barbara A |
author_facet | Watts, Lisa M Browne, John A Murphy, Barbara A |
author_sort | Watts, Lisa M |
collection | PubMed |
description | BACKGROUND: A comprehensive understanding of the equine circadian clock involves the evaluation of circadian clock gene expression. A non-invasive and effective method for detecting equine clock gene expression has yet to be established. Currently, research surrounding this area has relied on collecting tissue biopsies or blood samples that can often be costly, time consuming and uncomfortable for the animal. METHODS: Five mares were individually stabled under a light–dark (LD) cycle that mimicked the external environmental photoperiod during a time of year corresponding with the vernal equinox. Hair follicles were collected every 4 h over a 24-h period by plucking hairs from the mane. RNA was extracted and quantitative (q) PCR assays were performed to determine temporal expression patterns for the core clock genes; ARNTL, CRY1, PER1, PER2, NR1D2 and the clock controlled gene, DBP. RESULTS: Repeated measures ANOVA for the clock gene transcripts PER1 and PER2 and the clock controlled gene, DBP, revealed significant variation in expression over time (p < .05, respectively). Cosinor analysis confirmed a significant 24-h temporal component for PER1 (p = .002) and DBP (p = .0033) and also detected rhythmicity for NR1D2 (p = .0331). CONCLUSIONS: We show that the extraction of RNA from equine hair follicle cells can identify the circadian 24 h oscillations of specific clock genes and a clock-controlled gene and therefore provide a valuable non-invasive method for evaluating the equine peripheral circadian clock. This method will serve as a useful tool for future evaluations of equine circadian rhythms and their response to environmental changes. |
format | Online Article Text |
id | pubmed-3514281 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-35142812012-12-05 Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells Watts, Lisa M Browne, John A Murphy, Barbara A J Circadian Rhythms Research BACKGROUND: A comprehensive understanding of the equine circadian clock involves the evaluation of circadian clock gene expression. A non-invasive and effective method for detecting equine clock gene expression has yet to be established. Currently, research surrounding this area has relied on collecting tissue biopsies or blood samples that can often be costly, time consuming and uncomfortable for the animal. METHODS: Five mares were individually stabled under a light–dark (LD) cycle that mimicked the external environmental photoperiod during a time of year corresponding with the vernal equinox. Hair follicles were collected every 4 h over a 24-h period by plucking hairs from the mane. RNA was extracted and quantitative (q) PCR assays were performed to determine temporal expression patterns for the core clock genes; ARNTL, CRY1, PER1, PER2, NR1D2 and the clock controlled gene, DBP. RESULTS: Repeated measures ANOVA for the clock gene transcripts PER1 and PER2 and the clock controlled gene, DBP, revealed significant variation in expression over time (p < .05, respectively). Cosinor analysis confirmed a significant 24-h temporal component for PER1 (p = .002) and DBP (p = .0033) and also detected rhythmicity for NR1D2 (p = .0331). CONCLUSIONS: We show that the extraction of RNA from equine hair follicle cells can identify the circadian 24 h oscillations of specific clock genes and a clock-controlled gene and therefore provide a valuable non-invasive method for evaluating the equine peripheral circadian clock. This method will serve as a useful tool for future evaluations of equine circadian rhythms and their response to environmental changes. BioMed Central 2012-10-05 /pmc/articles/PMC3514281/ /pubmed/23039139 http://dx.doi.org/10.1186/1740-3391-10-7 Text en Copyright ©2012 Watts et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Watts, Lisa M Browne, John A Murphy, Barbara A Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells |
title | Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells |
title_full | Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells |
title_fullStr | Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells |
title_full_unstemmed | Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells |
title_short | Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells |
title_sort | investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3514281/ https://www.ncbi.nlm.nih.gov/pubmed/23039139 http://dx.doi.org/10.1186/1740-3391-10-7 |
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