Identification of genes and proteins related to hair growth and analysis of alopecia inducing and preventing factor

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At first, we succeed to purify mRNA directly from hair root and outer root sheath. Using mRNA from the normal male scalp hair and the scalp hair from male-type alopecia, expression difference among mRNA species were analyzed by DNA tip analysis. We focused on difference of expression of keratin that is supposed to affect the quality of hair. We also focused on genes related to melanin synthesis like tyrosinase (TYR) and tyrosinase related protein 1 (TYRP1) because hair color changes were frequently observed in male-type alopecia. TYR is a rate-limiting enzyme of this cascade and TYRP1 is essential to maintain TYR function. On DNA tip analysis, hair keratin KA36 and keratin 25 showed abundant expression in normal hair. In alopecia pair, hair keratin acidic 5 (KRTA5) and basic 5 (KRTB5) as well as keratin 15 (KRT15) were abundantly expressed but TYR and TYRP I were not. Next, we tried to quantitate their expression using RT-PCR. Expression of KRTA5 and B5 was approx. 3.5 times as much and p-actin expression in normal hair, whereas that in alopecia hair was approx. 5.5 times. KRT15 expression in normal hair and alopecia hair was respectively approx. 0.5 times and 6.5 times as that of β-actin, viz. KRT expression in alopecia was more than 13 times higher than that in normal hair. TYR and TYRP1 expression was approx. 0.1 and 0.3 times as that of (-actin, respectively. It is not always observed in all alopecia hair but some alopecia hair showed almost null expression of them. Even in alopecia patient, hair obtained from the non-bald area displayed the expression pattern similar with normal hair. Thus, we consider that the analysis of these genes can show us the way to distinguish the area affected and non-affected by male-type alopecia and apply to the patent.