Shorter K, Vafaee T, Picksley SM, Randall VA
Department of Biomedical Sciences, University of Bradford, UNITED KINGDOM

Macrophage-stimulating protein (MSP) also known as hepatocyte growth factor-like protein (HGFL) or scatter factor 2 belongs to the hepatocyte growth factor/scatter factor family. MSP is synthesised predominantly in the liver and acts as an inflammatory cytokine and regulator of cell growth. McElwee et al recently investigated MSP in hair follicles, using immunohistochemistry, western blotting, in vitro and in vivo studies, concluding MSP was a modulator of hair growth. The aim of this study was to investigate whether the mRNA for MSP is actually produced by human hair follicles. Human skin samples were treated with the RNAlater(tm) stabilisation reagent to preserve the mRNA by inhibiting ribonucleases. Individual anagen hair follicles were isolated by micro-dissection, pooled and total RNA extracted. RNA quality was checked before further purification to isolate poly(A)RNA; DNase treatment removed any contaminating DNA, before cDNA synthesis by reverse transcription. Reverse transcription-polymerase chain reaction (RT-PCR) was carried out on hair follicle and rat liver (positive control) cDNA, using specifically designed primers to detect MSP expression in both species. RT-PCR reactions were also performed using primers for the highly expressed gene, ß-actin, to assess the cDNAs' quality. PCR products were separated by gel electrophoresis, checked for appropriate sizes, extracted and sequenced. The ß-actin gene was highly expressed in all samples, demonstrating cDNA quality. Both hair follicle and rat liver samples produced appropriate sized bands for MSP. Sequence analysis confirmed PCR products from both rat liver and human hair follicles correlated to the known MSP gene. These results demonstrate that human hair follicles express mRNA for MSP supporting a role for MSP as a paracrine regulator in human hair growth.