Isolating RNA from precursor and mature melanocytes from human vitiligo and normal skin using laser capture microdissection.

ABSTRACT: To characterize the gene expression profile of regenerated melanocytes in the narrow band UVB (NBUVB)-treated vitiligo epidermis and their precursors in the hair follicle, we present here a strategy of RNA isolation from in situ melanocytes using human frozen skin. We developed a rapid immunostaining protocol using the NKI-beteb antibody, which labels differentiated and precursor melanocytes, followed by fluorescent laser capture microdissection. This technique enabled the direct isolation, from melanocyte and adjacent keratinocyte populations, of satisfactory quality RNA that was successfully amplified and analysed by qRT-PCR. The melanocyte-specific gene transcripts TYR, DCT, TYRP1 and PMEL were significantly upregulated in our NBUVB-treated melanocyte samples as compared with the kerati

SUBMITTER: Goldstein NB 

PROVIDER: S-EPMC5322465 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature