FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Circadian Rhythms / Investigation of a non-invasive method of assessing the equi...
Journal of circadian rhythms2012; 10(1); 7; doi: 10.1186/1740-3391-10-7

Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells.

Abstract: 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.
Get Full Text
Publication Date: 2012-10-05 PubMed ID: 23039139PubMed Central: PMC3514281DOI: 10.1186/1740-3391-10-7Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article is about the development and testing of a non-invasive method to study gene expression related to the circadian clock in horses using hair follicle cells.

Methodology

The study involved:

  • Five mares. They were individually kept in stables under a light-dark cycle that imitated the changing photoperiods of the vernal equinox.
  • Every 4 hours over a 24-hour cycle, hair follicles were collected from each horse by plucking hair from their mane.
  • RNA was extracted from these follicles and quantitative PCR assays were conducted to determine the temporal expression patterns of a set of genes related to the circadian clock: ARNTL, CRY1, PER1, PER2, and NR1D2, as well as DBP, a gene controlled by the circadian clock.

Results

The results indicated:

  • Significant variations in the expression of the clock gene transcripts PER1 and PER2, and the clock-controlled gene DBP, over time.
  • Cosinor analysis confirmed a 24-hour temporal component for PER1 and DBP and detected rhythm for NR1D2.
  • In effect, these results indicate that they were able to identify 24-hour oscillations of specific clock genes, and a clock-controlled gene, from RNA extracted from horse hair follicles.

Conclusions and Implications

  • The researchers concluded that the method they used is a valuable, non-invasive way to study the circadian clocks in horses.
  • This method will serve as a useful tool for future studies exploring equine circadian rhythms and their response to environmental changes.
  • Given that previous methods required tissue biopsies or blood samples, which are invasive, costly, time-consuming, and potentially uncomfortable for the animal, this novel method represents a significant improvement.

Cite This Article

APA
Watts LM, Browne JA, Murphy BA. (2012). Investigation of a non-invasive method of assessing the equine circadian clock using hair follicle cells. J Circadian Rhythms, 10(1), 7. https://doi.org/10.1186/1740-3391-10-7

Publication

Journal of circadian rhythms
ISSN: 1740-3391
NlmUniqueID: 101200389
Country: England
Language: English
Volume: 10
Issue: 1
Pages: 7

Researcher Affiliations

Watts, Lisa M
  • School of Agriculture and Food Science, University College Dublin, Belfield, Dublin 4, Ireland. barbara.murphy@ucd.ie.
Browne, John A
    Murphy, Barbara A

      References

      This article includes 27 references
      1. Murphy BA. Chronobiology and the horse: recent revelations and future directions.. Vet J 2010 Aug;185(2):105-14.
        doi: 10.1016/j.tvjl.2009.04.013pubmed: 19427248google scholar: lookup
      2. Lowrey PL, Takahashi JS. Mammalian circadian biology: elucidating genome-wide levels of temporal organization.. Annu Rev Genomics Hum Genet 2004;5:407-41.
        doi: 10.1146/annurev.genom.5.061903.175925pmc: PMC3770722pubmed: 15485355google scholar: lookup
      3. Reppert SM, Weaver DR. Coordination of circadian timing in mammals.. Nature 2002 Aug 29;418(6901):935-41.
        doi: 10.1038/nature00965pubmed: 12198538google scholar: lookup
      4. Ko CH, Takahashi JS. Molecular components of the mammalian circadian clock.. Hum Mol Genet 2006 Oct 15;15 Spec No 2:R271-7.
        doi: 10.1093/hmg/ddl207pubmed: 16987893google scholar: lookup
      5. Gekakis N, Staknis D, Nguyen HB, Davis FC, Wilsbacher LD, King DP, Takahashi JS, Weitz CJ. Role of the CLOCK protein in the mammalian circadian mechanism.. Science 1998 Jun 5;280(5369):1564-9.
        doi: 10.1126/science.280.5369.1564pubmed: 9616112google scholar: lookup
      6. Sato TK, Panda S, Miraglia LJ, Reyes TM, Rudic RD, McNamara P, Naik KA, FitzGerald GA, Kay SA, Hogenesch JB. A functional genomics strategy reveals Rora as a component of the mammalian circadian clock.. Neuron 2004 Aug 19;43(4):527-37.
        doi: 10.1016/j.neuron.2004.07.018pubmed: 15312651google scholar: lookup
      7. Panda S, Antoch MP, Miller BH, Su AI, Schook AB, Straume M, Schultz PG, Kay SA, Takahashi JS, Hogenesch JB. Coordinated transcription of key pathways in the mouse by the circadian clock.. Cell 2002 May 3;109(3):307-20.
        doi: 10.1016/S0092-8674(02)00722-5pubmed: 12015981google scholar: lookup
      8. Benloucif S, Guico MJ, Reid KJ, Wolfe LF, L'hermite-Balériaux M, Zee PC. Stability of melatonin and temperature as circadian phase markers and their relation to sleep times in humans.. J Biol Rhythms 2005 Apr;20(2):178-88.
        doi: 10.1177/0748730404273983pubmed: 15834114google scholar: lookup
      9. Refinetti R, Menaker M. The circadian rhythm of body temperature.. Physiol Behav 1992 Mar;51(3):613-37.
        doi: 10.1016/0031-9384(92)90188-8pubmed: 1523238google scholar: lookup
      10. Murphy BA, Elliott JA, Sessions DR, Vick MM, Kennedy EL, Fitzgerald BP. Rapid phase adjustment of melatonin and core body temperature rhythms following a 6-h advance of the light/dark cycle in the horse.. J Circadian Rhythms 2007 Aug 24;5:5.
        doi: 10.1186/1740-3391-5-5pmc: PMC2020455pubmed: 17718919google scholar: lookup
      11. Boivin DB, James FO, Wu A, Cho-Park PF, Xiong H, Sun ZS. Circadian clock genes oscillate in human peripheral blood mononuclear cells.. Blood 2003 Dec 1;102(12):4143-5.
        doi: 10.1182/blood-2003-03-0779pubmed: 12893774google scholar: lookup
      12. Bjarnason GA, Jordan RC, Wood PA, Li Q, Lincoln DW, Sothern RB, Hrushesky WJ, Ben-David Y. Circadian expression of clock genes in human oral mucosa and skin: association with specific cell-cycle phases.. Am J Pathol 2001 May;158(5):1793-801.
        doi: 10.1016/S0002-9440(10)64135-1pmc: PMC1891949pubmed: 11337377google scholar: lookup
      13. Akashi M, Soma H, Yamamoto T, Tsugitomi A, Yamashita S, Yamamoto T, Nishida E, Yasuda A, Liao JK, Node K. Noninvasive method for assessing the human circadian clock using hair follicle cells.. Proc Natl Acad Sci U S A 2010 Aug 31;107(35):15643-8.
        doi: 10.1073/pnas.1003878107pmc: PMC2932591pubmed: 20798039google scholar: lookup
      14. Martin AM, Elliott JA, Duffy P, Blake CM, Ben Attia S, Katz LM, Browne JA, Gath V, McGivney BA, Hill EW, Murphy BA. Circadian regulation of locomotor activity and skeletal muscle gene expression in the horse.. J Appl Physiol (1985) 2010 Nov;109(5):1328-36.
        doi: 10.1152/japplphysiol.01327.2009pubmed: 20847133google scholar: lookup
      15. Refinetti R, Lissen GC, Halberg F. Procedures for numerical analysis of circadian rhythms.. Biol Rhythm Res 2007;38(4):275-325.
        doi: 10.1080/09291010600903692pmc: PMC3663600pubmed: 23710111google scholar: lookup
      16. Halberg F, Tong YL, Johnson EA. Circadian system phase, an aspect of temporal morphology: procedures and illustrative examples. The Cellular Aspects of Biorhythms 1967; pp. 22–48.
      17. Piccione G, Caola G, Refinetti R. The circadian rhythm of body temperature of the horse. Biological Rhythm Research 2002;33:113–119.1.
        doi: 10.1076/brhm.33.1.113.1322google scholar: lookup
      18. McGlynn OF, Browne JA, Blake CM, Murphy BA. Time of day influences cytokine and clock gene response to immune stimulation in equine whole blood. Anim Genet 2010;41:202–204.
      19. Kolla BP, Auger RR. Jet lag and shift work sleep disorders: how to help reset the internal clock.. Cleve Clin J Med 2011 Oct;78(10):675-84.
        doi: 10.3949/ccjm.78a.10083pubmed: 21968474google scholar: lookup
      20. Krystal AD. How the circadian rhythm affects sleep, wakefulness, and overall health: background for understanding shift work disorder.. J Clin Psychiatry 2012 Feb;73(2):e05.
        doi: 10.4088/JCP.11073br1pubmed: 22401482google scholar: lookup
      21. Konturek PC, Brzozowski T, Konturek SJ. Gut clock: implication of circadian rhythms in the gastrointestinal tract.. J Physiol Pharmacol 2011 Apr;62(2):139-50.
        pubmed: 21673361
      22. Boyce P, Barriball E. Circadian rhythms and depression.. Aust Fam Physician 2010 May;39(5):307-10.
        pubmed: 20485718
      23. McCarthy JJ, Andrews JL, McDearmon EL, Campbell KS, Barber BK, Miller BH, Walker JR, Hogenesch JB, Takahashi JS, Esser KA. Identification of the circadian transcriptome in adult mouse skeletal muscle.. Physiol Genomics 2007 Sep 19;31(1):86-95.
        doi: 10.1152/physiolgenomics.00066.2007pmc: PMC6080860pubmed: 17550994google scholar: lookup
      24. Yan J, Wang H, Liu Y, Shao C. Analysis of gene regulatory networks in the mammalian circadian rhythm.. PLoS Comput Biol 2008 Oct;4(10):e1000193.
        doi: 10.1371/journal.pcbi.1000193pmc: PMC2543109pubmed: 18846204google scholar: lookup
      25. Reddy AB, Field MD, Maywood ES, Hastings MH. Differential resynchronisation of circadian clock gene expression within the suprachiasmatic nuclei of mice subjected to experimental jet lag.. J Neurosci 2002 Sep 1;22(17):7326-30.
        pmc: PMC6757999pubmed: 12196553doi: 10.1523/jneurosci.22-17-07326.2002google scholar: lookup
      26. Nagano M, Adachi A, Nakahama K, Nakamura T, Tamada M, Meyer-Bernstein E, Sehgal A, Shigeyoshi Y. An abrupt shift in the day/night cycle causes desynchrony in the mammalian circadian center.. J Neurosci 2003 Jul 9;23(14):6141-51.
        pmc: PMC6740348pubmed: 12853433doi: 10.1523/jneurosci.23-14-06141.2003google scholar: lookup
      27. Yamazaki S, Numano R, Abe M, Hida A, Takahashi R, Ueda M, Block GD, Sakaki Y, Menaker M, Tei H. Resetting central and peripheral circadian oscillators in transgenic rats.. Science 2000 Apr 28;288(5466):682-5.
        doi: 10.1126/science.288.5466.682pubmed: 10784453google scholar: lookup

      Citations

      This article has been cited 6 times.
      1. Collery A, Browne JA, O'Brien C, Sheridan JT, Murphy BA. Optimised Stable Lighting Strengthens Circadian Clock Gene Rhythmicity in Equine Hair Follicles.. Animals (Basel) 2023 Jul 17;13(14).
        doi: 10.3390/ani13142335pubmed: 37508112google scholar: lookup
      2. Kudlova N, Slavik H, Duskova P, Furst T, Srovnal J, Bartek J, Mistrik M, Hajduch M. An efficient, non-invasive approach for in-vivo sampling of hair follicles: design and applications in monitoring DNA damage and aging.. Aging (Albany NY) 2021 Dec 6;13(23):25004-25024.
        doi: 10.18632/aging.203744pubmed: 34874896google scholar: lookup
      3. Zhang CZ, Sun HZ, Li SL, Sang D, Zhang CH, Jin L, Antonini M, Zhao CF. Effects of photoperiod on nutrient digestibility, hair follicle activity and cashmere quality in Inner Mongolia white cashmere goats.. Asian-Australas J Anim Sci 2019 Apr;32(4):541-547.
        doi: 10.5713/ajas.18.0154pubmed: 30145869google scholar: lookup
      4. Ferrante A, Gellerman D, Ay A, Woods KP, Filipowicz AM, Jain K, Bearden N, Ingram KK. Diurnal Preference Predicts Phase Differences in Expression of Human Peripheral Circadian Clock Genes.. J Circadian Rhythms 2015 Jun 5;13:4.
        doi: 10.5334/jcr.aepubmed: 27103930google scholar: lookup
      5. Plikus MV, Van Spyk EN, Pham K, Geyfman M, Kumar V, Takahashi JS, Andersen B. The circadian clock in skin: implications for adult stem cells, tissue regeneration, cancer, aging, and immunity.. J Biol Rhythms 2015 Jun;30(3):163-82.
        doi: 10.1177/0748730414563537pubmed: 25589491google scholar: lookup
      6. Sato M, Matsuo T, Atmore H, Akashi M. Possible contribution of chronobiology to cardiovascular health.. Front Physiol 2013;4:409.
        doi: 10.3389/fphys.2013.00409pubmed: 24478711google scholar: lookup
      Subscribe to our newsletter
      Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.