Secretion of corticotropin-releasing hormone by superfused human placental fragments

Abstract

Corticotropin-releasing hormone (CRH) immunoreactivity (IR) is present in the blood of women in the 3rd trimester of pregnancy and in placental extracts. We have used a placental fragment superfusion system to investigate the release of CRH from fresh placental tissue. Fragments of normal term placenta were mixed with Biogel P2, packed into minicolumns and superfused with carbogen-gassed Earles buffer at 37 degrees C. The rheology of the superfusion system was determined and the oxygen consumption of the superfused placental fragments indicated viability of the tissue preparation over a 5-hour time span. CRH IR in the eluate was measured by radioimmunoassay (RIA) using the 41 residue synthetic peptide human, rat CRH-41 (h, r CRH-41) as the standard, 125I labelled Tyr- h, r CRH as the tracer and rabbit anti-ovine CRH as the antibody. The sensitivity of the assay is 2 pM. Size exclusion chromatography on Sephadex G-50 of the placental column eluate displayed one major peak of CRH IR which co-eluted with that of h, r CRH. Placental fragment superfusate displayed potent CRH bioactivity as assessed by beta-endorphin secretion from ovine pituitary cells. Replacing the superfusing medium of the placental fragments with 45 mM KCl resulted in a prompt increase in the release of CRH IR. These results indicate that placental cells in vitro secrete a molecule of similar molecular weight, immunoreactivity and bioactivity to h, r CRH and that the rate of secretion may be regulated.