Comparison of suction blistering and tape stripping for analysis of epidermal genes, proteins and lipids
Jazyk angličtina Médium print-electronic
Typ dokumentu srovnávací studie, časopisecké články
PubMed
28905096
DOI
10.1007/s00403-017-1776-6
PII: 10.1007/s00403-017-1776-6
Knihovny.cz E-zdroje
- Klíčová slova
- Confocal microscopy, Epidermis, Stratum corneum, Suction blistering, Tape stripping,
- MeSH
- chromatografie na tenké vrstvě MeSH
- claudin-1 analýza MeSH
- dospělí MeSH
- epidermis chemie MeSH
- filagriny MeSH
- imunohistochemie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé středního věku MeSH
- lidé MeSH
- lipidy analýza MeSH
- messenger RNA analýza MeSH
- odsávání MeSH
- proteiny intermediálních filament analýza MeSH
- proteiny analýza MeSH
- puchýř MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- claudin-1 MeSH
- CLDN1 protein, human MeSH Prohlížeč
- filagriny MeSH
- FLG protein, human MeSH Prohlížeč
- lipidy MeSH
- messenger RNA MeSH
- proteiny intermediálních filament MeSH
- proteiny MeSH
Analysis of epidermal genes, proteins and lipids is important in the research and diagnosis of skin diseases. Although punch biopsy is the first-choice technique for the skin sampling, it is unnecessarily invasive for obtaining a sample just for the epidermal analysis. Here we compare two less invasive methods, suction blistering (SB) and tape stripping (TS), for the analysis of selected epidermal genes (quantitative real-time reverse transcription PCR, qRT-PCR), proteins (western blotting, WB), and lipids in ten healthy volunteers. TS provided significantly less material than SB and no viable epidermal layers could be obtained according to the reflectance confocal microscopy. Consistently, only the SC protein filaggrin and housekeeping GAPDH together with FLG and RPL13A mRNA were detected by TS. In the SB samples, WB and qRT-PCR could easily detect all the selected proteins (claudin-1, occludin, filaggrin, laminin and GAPDH) and genes (CLDN1, OCLN, FLG, LAMA3 and RPL13A), respectively. A single SB sample further provided enough of material for immunohistochemistry and lipid analyses, which was not feasible with the TS samples. Immunohistochemistry of the SB samples showed intact epidermal structure and a characteristic expression of claudin-1. Infrared spectroscopy showed well-ordered lipids with both orthorhombic and hexagonal packing and high-performance thin layer chromatography confirmed all lipid classes (including ceramide subclasses) in correct proportions. Taken together, SB represents a reliable sampling technique that can be utilized for multipurpose epidermal analyses in various studies.
Department of Research and Development Contipro a s Dolní Dobrouč Czech Republic
Skin Barrier Research Group Faculty of Pharmacy Charles University Hradec Králové Czech Republic
Citace poskytuje Crossref.org
