Endothelin receptor B expression in the rat and rabbit lung as determined by in situ hybridization using nonisotopic probes.
Endothelins are a family of potent vasoactive peptides. Full-length cDNA clones to human endothelin receptor B (ETB) mRNA were random prime-labeled with nucleotides conjugated to digoxigenin for in situ hybridization. The labeled cDNA was used to probe frozen sections of rat and rabbit lung. Detection of the digoxigenin-labeled probe was accomplished by an antibody-enzyme conjugate, anti-digoxigenin alkaline phosphatase. The location of the antibody-antigen complex was visualized as an enzyme-linked color reaction. The hybridization, washings, and detection steps were performed under stringent conditions. The following cell types of the rat and rabbit lung had abundant positive reaction product to the ETB probe: bronchiolar and bronchial epithelium, endothelium of smooth-muscle--walled vessels, and bronchial and bronchiolar-associated lymphoid tissue. Abundant positive reaction product was also observed in cell populations in the lung parenchyma. Additional studies are being performed to identify those populations. The results of this study suggest that in addition to vasoactivity, endothelins play other important roles in the lung.
Durham
S K
SK
Department of Experimental Pathology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ 08543.
Goller
N L
NL
Lynch
J S
JS
Fisher
S M
SM
Rose
P M
PM
D000818:Animals; D000936:Antigen-Antibody Complex / Q000276:immunology / Q000378:metabolism; D015342:DNA Probes; D018076:DNA, Complementary / Q000378:metabolism; D004076:Digoxigenin / Q000276:immunology; D006801:Humans; D017403:In Situ Hybridization; D008168:Lung / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D051381:Rats; D017466:Receptors, Endothelin / Q000096:biosynthesis*
D000936:Antigen-Antibody Complex; D015342:DNA Probes; D018076:DNA, Complementary; D012333:RNA, Messenger; D017466:Receptors, Endothelin; D004076:Digoxigenin