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Article

Identification of Reference Genes for Circadian Studies on Brain Microvessels and Choroid Plexus Samples Isolated from Rats

1
Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland
2
Institute of Biophysics, Biological Research Centre, 6726 Szeged, Hungary
*
Authors to whom correspondence should be addressed.
Academic Editors: Randy J. Nelson, James C. Walton and Vladimir N. Uversky
Biomolecules 2021, 11(8), 1227; https://doi.org/10.3390/biom11081227
Received: 3 July 2021 / Revised: 11 August 2021 / Accepted: 13 August 2021 / Published: 17 August 2021
(This article belongs to the Special Issue Therapeutic Aspects of Circadian Rhythms)
Delivery of putative compounds of therapeutic value to the brain is limited by brain barriers: the blood–brain barrier located in the endothelium of the brain microvessels (BrMVs) and the blood–cerebrospinal fluid barrier located in the epithelium of the choroid plexus (ChP). Understanding their function and modulation by the circadian clock may enhance the efficacy of brain-targeting therapies. The aim of the present study was to evaluate the stability of 10 reference genes in the BrMV and ChP, isolated from male and female rats at six time points (ZT1, 5, 9, 13, 17, and 21). Gene evaluations were performed by qPCR, analyzed by RefFinder tool, and verified by analyzing the expression of the brain and muscle ARNT-like 1 (Bmal1) using the qPCR and digital PCR methods. We identified as the most stable genes for circadian studies tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (Ywhaz) and apolipoprotein E (Apoe) for BrMV, and beta actin (Actb) and hypoxanthine-guanine phosphoribosyltransferase (Hprt1) for ChP. After verification, ribosomal protein (Rps18) was also included as a sufficient reference gene. Additionally, the observed gender difference in the Bmal1 oscillations in both BrMV and ChP suggests that separate studies for each gender are recommended. View Full-Text
Keywords: reference genes; circadian rhythm; brain barriers; brain microvessels; choroid plexus; qPCR reference genes; circadian rhythm; brain barriers; brain microvessels; choroid plexus; qPCR
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MDPI and ACS Style

Szczepkowska, A.; Harazin, A.; Barna, L.; Deli, M.A.; Skipor, J. Identification of Reference Genes for Circadian Studies on Brain Microvessels and Choroid Plexus Samples Isolated from Rats. Biomolecules 2021, 11, 1227. https://doi.org/10.3390/biom11081227

AMA Style

Szczepkowska A, Harazin A, Barna L, Deli MA, Skipor J. Identification of Reference Genes for Circadian Studies on Brain Microvessels and Choroid Plexus Samples Isolated from Rats. Biomolecules. 2021; 11(8):1227. https://doi.org/10.3390/biom11081227

Chicago/Turabian Style

Szczepkowska, Aleksandra, András Harazin, Lilla Barna, Mária A. Deli, and Janina Skipor. 2021. "Identification of Reference Genes for Circadian Studies on Brain Microvessels and Choroid Plexus Samples Isolated from Rats" Biomolecules 11, no. 8: 1227. https://doi.org/10.3390/biom11081227

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