Nestin in human Skin: exclusive expression in intramesenchymal skin compartments and regulation by leptin

TY - JOUR

T1 - Nestin in human Skin: exclusive expression in intramesenchymal skin compartments and regulation by leptin

AU - Tiede, Stephan

AU - Kloepper, Jennifer E

AU - Ernst, Nancy

AU - Poeggeler, Burkard

AU - Kruse, Charli

AU - Paus, Ralf

PY - 2009/11

Y1 - 2009/11

N2 - Cutaneous nestin+ cells are of substantial interest in regenerative medicine. However, the location of nestin+ cells in situ remains controversial. We therefore sought to determine their location in female human scalp skin, using stringently controlled immunohistochemical techniques, Western blot analysis, and in situ hybridization and complementing those techniques with relative and quantitative reverse transcriptase-PCR of enzymatically digested or laser-capture microdissected human hair follicle (HF) compartments. We show here that the immunoreactivity (IR) patterns obtained with anti-nestin antibodies are highly dependent on the tissue-fixation and immunohistochemical methods used. NESTIN mRNA could not be detected within HF-associated epithelial cells in situ or in RNA extracts of the microdissected HF epithelium. Instead, NESTIN transcripts were found only in intramesenchymal skin compartments. Individual cells showing both, specific nestin IR and NESTIN mRNA were detectable in the connective-tissue sheaths of human HFs, sebaceous and sweat glands. Moreover, stimulation of organ-cultured human scalp skin with the adipokine leptin increased the number of nestin+ cells in these intramesenchymal skin locations, whereas no specific nestin IR could be induced by leptin within the HF epithelium, including the bulge. Therefore, nestin expression at the gene and protein levels in human scalp skin is restricted to the periappendage mesenchyme and can be stimulated by leptin.

AB - Cutaneous nestin+ cells are of substantial interest in regenerative medicine. However, the location of nestin+ cells in situ remains controversial. We therefore sought to determine their location in female human scalp skin, using stringently controlled immunohistochemical techniques, Western blot analysis, and in situ hybridization and complementing those techniques with relative and quantitative reverse transcriptase-PCR of enzymatically digested or laser-capture microdissected human hair follicle (HF) compartments. We show here that the immunoreactivity (IR) patterns obtained with anti-nestin antibodies are highly dependent on the tissue-fixation and immunohistochemical methods used. NESTIN mRNA could not be detected within HF-associated epithelial cells in situ or in RNA extracts of the microdissected HF epithelium. Instead, NESTIN transcripts were found only in intramesenchymal skin compartments. Individual cells showing both, specific nestin IR and NESTIN mRNA were detectable in the connective-tissue sheaths of human HFs, sebaceous and sweat glands. Moreover, stimulation of organ-cultured human scalp skin with the adipokine leptin increased the number of nestin+ cells in these intramesenchymal skin locations, whereas no specific nestin IR could be induced by leptin within the HF epithelium, including the bulge. Therefore, nestin expression at the gene and protein levels in human scalp skin is restricted to the periappendage mesenchyme and can be stimulated by leptin.

KW - Antibody Specificity

KW - Blotting, Western

KW - Epithelial Cells

KW - Female

KW - Hair Follicle

KW - Humans

KW - Immunohistochemistry

KW - In Situ Hybridization

KW - Intermediate Filament Proteins

KW - Leptin

KW - Mesenchymal Stromal Cells

KW - Middle Aged

KW - Nerve Tissue Proteins

KW - Nestin

KW - Organ Culture Techniques

KW - Scalp

KW - Sebaceous Glands

KW - Sweat Glands

KW - Tissue Fixation

KW - Transcription, Genetic

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1038/jid.2009.148

DO - 10.1038/jid.2009.148

M3 - SCORING: Journal article

C2 - 19554024

VL - 129

SP - 2711

EP - 2720

JO - J INVEST DERMATOL

JF - J INVEST DERMATOL

SN - 0022-202X

IS - 11

ER -