Abstract
The expression of glypicans in different hair follicle (HF) compartments and their potential roles during hair shaft growth are still poorly understood. Heparan sulfate proteoglycan (HSPG) distribution in HFs is classically investigated by conventional histology, biochemical analysis, and immunohistochemistry. In this report, a novel approach is proposed to assess hair histology and HSPG distribution changes in HFs at different phases of the hair growth cycle using infrared spectral imaging (IRSI). The distribution of HSPGs in HFs was probed by IRSI using the absorption region relevant to sulfation as a spectral marker. The findings were supported by Western immunoblotting and immunohistochemistry assays focusing on the glypican-1 expression and distribution in HFs. This study demonstrates the capacity of IRSI to identify the different HF tissue structures and to highlight protein, proteoglycan (PG), glycosaminoglycan (GAG), and sulfated GAG distribution in these structures. The comparison between anagen, catagen, and telogen phases shows the qualitative and/or quantitative evolution of GAGs as supported by Western immunoblotting. Thus, IRSI can simultaneously reveal the location of proteins, PGs, GAGs, and sulfated GAGs in HFs in a reagent- and label-free manner. From a dermatological point of view, IRSI shows its potential as a promising technique to study alopecia.
Highlights
We propose in this original study to probe the distribution of Heparan sulfate proteoglycan (HSPG) in hair follicle (HF) at different phases of the hair growth cycle by infrared spectral imaging (IRSI) using sulfation as a spectral marker
The present report describes the evolution of GAGs, sulfated GAGs, and, the heparan sulfate (HS)-type GAGs and GPC1 distribution in hair follicles at different phases of the hair growth cycle using spectral imaging
The protein, HSPG, and GAG contribution varied within HFs forming five different clusters corresponding to different parts of the HF (the connective tissue sheath (CTS), the ORS, the IRS, the bulb, and the hair shaft), independently of the phase of the hair growth cycle (Figure 4A–C)
Summary
Two major spectral ranges were used to characterize HSPG distribution: the spectral window 1800–900 cm−1 , known as the fingerprint region, shown to be the most specific range for GAG studies [24], and the spectral window 1350–1190 cm−1 , centered at 1248 cm−1 , that is specific for GAG sulfation [29] Based on this knowledge, we propose in this original study to probe the distribution of HSPGs in HFs at different phases of the hair growth cycle by IRSI using sulfation as a spectral marker. The goal was to study HSPG, GAG, and sulfated GAG distribution and variation in HFs without any staining or labeling It allows investigating these changes during the hair growth cycle. Our data were supported by Western immunoblotting and immunohistochemistry assays, by analyzing the GPC1 expression and distribution in HFs at different phases of the cycle
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