RNA purification from epidermal suction blisters.
Certain inflammatory skin diseases are accompanied by increased cytokine concentrations in the epidermis. To determine whether these cytokines are synthesized in the epidermis or exported from underlying tissues, epidermal RNA was analysed for the presence of their messenger RNAs. We report a method for RNA extraction from pure epidermal samples isolated by the suction blister method. The yield of total RNA was sufficient for hybridization experiments (12-38 micrograms per seven blisters, 5 mm in diameter). Using RNA extracted by this method, we have demonstrated the presence of messenger RNA for glyceraldehyde-3-phosphate-dehydrogenase in 13 preparations from suction blisters obtained from tuberculin skin reactions, positive patch test reactions, or normal skin. We did not, however, observe messenger RNA for interleukin 1 alpha or 8 in these preparations.
Department of Dermatology, University of Aarhus, Marselisborg Hospital, Denmark.
M
M
Kristensen
C G
CG
Larsen
P
P
Jørgensen
K
K
Paludan
D001768:Blister / Q000209:etiology / Q000378:metabolism*; D015152:Blotting, Northern / Q000379:methods; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis; D016209:Interleukin-8 / Q000096:biosynthesis; D015603:Keratinocytes / Q000378:metabolism*; D010328:Patch Tests; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D013396:Suction / Q000379:methods; D014374:Tuberculin Test
D007375:Interleukin-1; D016209:Interleukin-8; D012333:RNA, Messenger