Repulsive guidance molecule a expression in normal human skin cells

Summary: Abstract Body: Background：Repulsive guidance molecules (RGMs) gene family consists of four members, RGMa, RGMb, RGMc and RGMd. Previous studies have found that RGMs expression is limited to a few tissues and their biological functions are not well understood. RGMs are now known as key players in many fundamental processes, such as cell migration, differentiation, iron homeostasis and apoptosis, etc. However, the expression and function of RGMs in normal human skin cells has not been reported. Objective: The aim of the present study was to evaluate RGMs expression in human skin. Method：Human skin cell was derived from prepuce tissue discarded after surgery in children. Suspended melanocyte were obtained from children foreskin with two-step enzyme-digestion method and then cultured in M254 medium supplied with human melanocyte growth supplement for three passages. The keratinocyte was grown in EpiLife® medium with human keratinocyte growth supplement for three passages. The fibroblasts were harvested from human dermis tissues with the tissue explants adherent method and was cultured in DMEM supplemented with 10% fetal bovine serum supplement for three passages. The mRNA and protein levels of RGMs in melanocytes, keratinocytes and fibroblasts were detected by RT-qPCR and western blot analysis, respectively. The expression and localization of RGMs in these cells were detected with immunofluorescent staining under laser scanning confocal microscopy. Results: The results of RT-qPCR and western blot demonstrated that the expression of RGMa mRNA and protein were detected in melanocytes, keratinocytes and fibroblasts. Remarkably, the RGMb and RGMc mRNA and protein were not detected. RGMa was expressed and localized to the nucleus and membrane of melanocytes, keratinocytes and fibroblasts under laser scanning confocal microscopy. Conclusion: RGMa expression in normal human skin melanocytes, keratinocytes and fibroblasts and may play an important role in human skin physiological functions. Yinghua Lan¹, Yu Wang¹, Hongguang Lu¹ 1. Department of Dermatology, Affiliated Hospital of Guizhou Medical University, Guiyang, China. Translational Studies: Cell and Molecular Biology