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Article

Role of Methylation in Period2 (PER2) Transcription in the Context of the Presence or Absence of Light Signals: Natural and Chemical—Studies on the Pig Model

1
Department of Local Physiological Regulations, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland
2
School of Life Sciences, Chair of Livestock Biotechnology, Technical University of Munich, D-85354 Freising, Germany
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Department of Neurosurgery, School of Medicine, University of Warmia and Mazury, 10-719 Olsztyn, Poland
4
Department of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury, 10-719 Olsztyn, Poland
*
Author to whom correspondence should be addressed.
Academic Editor: Amelia Casamassimi
Int. J. Mol. Sci. 2021, 22(15), 7796; https://doi.org/10.3390/ijms22157796
Received: 19 May 2021 / Revised: 16 July 2021 / Accepted: 16 July 2021 / Published: 21 July 2021
It has been proposed that carbon monoxide (CO) is a chemical light carrier that is transferred by the humoral pathway from the retina to the brain. Here, we aimed to study how deeply CO is involved in regulating the expression of Period2 gene (PER2), one of the genes maintaining the intrinsic biological clock. In our in vivo experiment, we studied whether CO may be a chemical signal and is also equivalent to natural light in three groups of pigs: Normal: housed in natural conditions without any procedures, Control: adapted and kept in constant darkness, infused with blank plasma, and CO treated: adapted and kept in constant darkness infused with CO-enriched plasma. After the experiment, the animals were slaughtered at two times of day: 12 p.m. and 12 a.m. Next, hypothalamus samples were collected. Quantitative PCR, the DNA methylation of the promoter sequence containing enhancers (E-box) and a functional analysis of the PER2 promoter was performed. qPCR showed a differential pattern of PER2 mRNA expression at daytime oscillation in the examined groups. Pyrosequencing revealed daytime changes in the methylation level of regulatory sites of the examined sequence. Luciferase reporter assay confirmed that E-boxes (CANNTG) drive the expression of the porcine PER2 in vitro. In conclusion, changes in methylation over 24 h may regulate the oscillatory manner of PER2 expression. View Full-Text
Keywords: gene expression; E-box; methylation; PER2; carbon monoxide; oscillation gene expression; E-box; methylation; PER2; carbon monoxide; oscillation
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MDPI and ACS Style

Gilun, P.; Flisikowski, K.; Flisikowska, T.; Kwiatkowska, J.; Wąsowska, B.; Koziorowska-Gilun, M. Role of Methylation in Period2 (PER2) Transcription in the Context of the Presence or Absence of Light Signals: Natural and Chemical—Studies on the Pig Model. Int. J. Mol. Sci. 2021, 22, 7796. https://doi.org/10.3390/ijms22157796

AMA Style

Gilun P, Flisikowski K, Flisikowska T, Kwiatkowska J, Wąsowska B, Koziorowska-Gilun M. Role of Methylation in Period2 (PER2) Transcription in the Context of the Presence or Absence of Light Signals: Natural and Chemical—Studies on the Pig Model. International Journal of Molecular Sciences. 2021; 22(15):7796. https://doi.org/10.3390/ijms22157796

Chicago/Turabian Style

Gilun, Przemysław, Krzysztof Flisikowski, Tatiana Flisikowska, Joanna Kwiatkowska, Barbara Wąsowska, and Magdalena Koziorowska-Gilun. 2021. "Role of Methylation in Period2 (PER2) Transcription in the Context of the Presence or Absence of Light Signals: Natural and Chemical—Studies on the Pig Model" International Journal of Molecular Sciences 22, no. 15: 7796. https://doi.org/10.3390/ijms22157796

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