In situ hybridization of atrial natriuretic peptide mRNA in the endothelial cells of human umbilical vessels.
The localization of mRNA encoding preproatrial natriuretic peptide (ANP) was investigated in cultured human umbilical vein endothelial cells (HUVEC) and tissue preparations of umbilical vein and artery. The techniques used were in situ hybridization and in situ hybridization combined with immunocytochemistry, using 32P-radiolabelled and non-radioactive digoxigenin labelled complementary RNA probes. Human ANP mRNAs are mainly localized in the endothelial cells of the umbilical vein and, to a lesser extent, in the endothelial cells of the umbilical artery. The autoradiographic labelling and the intensity of digoxigenin staining were significantly reduced by treatment with RNase before in situ hybridization. This study provides unequivocal evidence for the expression of the ANP gene in the endothelial cells of human umbilical vessels, confirming that these endothelial cells have the ability to synthesize this peptide. The functional significance of the presence of the ANP mRNA in the endothelial cells of human umbilical vessels is discussed.
Cai
W Q
WQ
Department of Anatomy and Developmental Biology, University College London, UK.
Terenghi
G
G
Bodin
P
P
Burnstock
G
G
Polak
J M
JM
D009320:Atrial Natriuretic Factor / Q000096:biosynthesis*; D002478:Cells, Cultured; D004076:Digoxigenin / Q000276:immunology / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D015347:RNA Probes; D012333:RNA, Messenger / Q000378:metabolism*; D012260:Ribonucleases / Q000276:immunology / Q000378:metabolism; D014469:Umbilical Arteries / Q000166:cytology / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism
D015347:RNA Probes; D012333:RNA, Messenger; D009320:Atrial Natriuretic Factor; D012260:Ribonucleases; D004076:Digoxigenin