Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Watts, Lisa M, Browne, John A, Murphy, Barbara A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
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
_version_ 1782252006530875392
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
work_keys_str_mv AT wattslisam investigationofanoninvasivemethodofassessingtheequinecircadianclockusinghairfolliclecells
AT brownejohna investigationofanoninvasivemethodofassessingtheequinecircadianclockusinghairfolliclecells
AT murphybarbaraa investigationofanoninvasivemethodofassessingtheequinecircadianclockusinghairfolliclecells