Universität zu Köln

Etich, Julia ORCID: 0000-0003-3238-6692, Bergmeier, Vera, Pitzler, Lena and Brachvogel, Bent (2017). Identification of a reference gene for the quantification of mRNA and miRNA expression during skin wound healing. Connect. Tissue Res., 58 (2). S. 196 - 208. PHILADELPHIA: TAYLOR & FRANCIS INC. ISSN 1607-8438

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Abstract

Aim: Wound healing is a coordinated process to restore tissue homeostasis and reestablish the protective barrier of the skin. miRNAs may modulate the expression of target genes to contribute to repair processes, but due to the complexity of the tissue it is challenging to quantify gene expression during the distinct phases of wound repair. Here, we aimed to identify a common reference gene to quantify changes in miRNA and mRNA expression during skin wound healing. Methods: Quantitative real-time PCR and bioinformatic analysis tools were used to identify suitable reference genes during skin repair and their reliability was tested by studying the expression of mRNAs and miRNAs. Results: Morphological assessment of wounds showed that the injury model recapitulates the distinct phases of skin repair. Non-degraded RNA could be isolated from skin and wounds and used to study the expression of non-coding small nuclear RNAs during wound healing. Among those, RNU6B was most constantly expressed during skin repair. Using this reference gene we could confirm the transient upregulation of IL-1 and PTPRC/CD45 during the early phase as well as the increased expression of collagen type I at later stages of repair and validate the differential expression of miR-204, miR-205, and miR-31 in skin wounds. In contrast to Gapdh the normalization to multiple reference genes gave a similar outcome. Conclusion:RNU6B is an accurate alternative normalizer to quantify mRNA and miRNA expression during the distinct phases of skin wound healing when analysis of multiple reference genes is not feasible.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Etich, Julia UNSPECIFIED orcid.org/0000-0003-3238-6692 UNSPECIFIED Bergmeier, Vera UNSPECIFIED UNSPECIFIED UNSPECIFIED Pitzler, Lena UNSPECIFIED UNSPECIFIED UNSPECIFIED Brachvogel, Bent UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-235711
DOI:	10.1080/03008207.2016.1210606
Journal or Publication Title:	Connect. Tissue Res.
Volume:	58
Number:	2
Page Range:	S. 196 - 208
Date:	2017
Publisher:	TAYLOR & FRANCIS INC
Place of Publication:	PHILADELPHIA
ISSN:	1607-8438
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language RELATIVE QUANTIFICATION; VASCULAR INTEGRITY; TARGET RECOGNITION; GRANULATION-TISSUE; HOUSEKEEPING GENE; III COLLAGEN; PCR DATA; RT-PCR; FIBROBLASTS; MICRORNAS Multiple languages Cell Biology; Orthopedics Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/23571