Analysis of Gene Expression in Isolated Single Hair Follicles Using Semiquantitative Reverse-Transcriptase Polymerase Chain Reaction
May 1995
in “
Journal of Investigative Dermatology
”
gene expression single hair follicles semequantitative reverse-transcriptase-polymerase chain reaction RT-PCR intercellular adhesion molecule ICAM-1 interferon IFN-γ anagen hair follicles RNA extraction reverse transcription PCR amplification specific primers β-actin internal standard high-performance liquid chromatography HPLC hair biology pathophysiology of hair growth gene expression single hair follicles RT-PCR ICAM-1 interferon IFN-γ anagen hair follicles RNA extraction reverse transcription PCR amplification specific primers β-actin internal standard HPLC hair biology pathophysiology of hair growth
TLDR Researchers developed a new way to measure gene activity in single hair follicles and found that a specific gene's activity changes with different amounts and times of treatment.
In 1995, researchers established a novel in vitro assay for the semiquantitative measurement of gene expression in single hair follicles using semiquantitative reverse-transcriptase-polymerase chain reaction (RT-PCR). They demonstrated the method by analyzing the expression of intercellular adhesion molecule (ICAM)-1 mRNA in hair follicles cultured with interferon (IFN)-γ. Isolated anagen hair follicles were cultured, and after incubation, RNA was extracted, reverse transcribed, and amplified using specific primers for ICAM-1 and β-actin (as an internal standard). The PCR products were quantified using high-performance liquid chromatography (HPLC). The study found a concentration and time dependency of IFN-γ-induced ICAM-1 expression in individual hair follicles, with peak expression after 7 hours of stimulation with 100 U/ml IFN-γ. This method provided a tool to study various aspects of hair biology and could offer new insights into the biology and pathophysiology of hair growth.
