The fate of homologous 125I-labelled immunoglobulin G within rat visceral yolk sacs incubated in vitro.
When 125I-labelled rat IgG (immunoglobulin G) is incubated in vitro with visceral yolk sacs from 17.5-day-pregnant rats, the protein is readily degraded. The major radioactive digestion product that accumulates in the medium is [125I]iodo-L-tyrosine. When rotenone (10 microM) is also present in the incubation medium, the rate of digestion of IgG is inhibited to the same extent as the rate of pinocytosis of 125I-labelled polyvinylpyrrolidone. Proteolysis is likewise inhibited when either NH4Cl (30 mM) or leupeptin (30 micrograms/ml) is present in the medium. The above findings strongly suggest that the observed proteolysis occurs within lysosomes. Normally, yolk sacs that have been exposed in vitro to radiolabelled substrates release radioactivity slowly when the tissue is re-incubated, unless the substrate can be degraded within lysosomes and released in the form of low-molecular-weight hydrolysis products. However, in such experiments 125I-labelled rat IgG shows quite exceptional behaviour in being rapidly released in an apparently intact form (as well as being degraded). If an agent that inhibits pinocytosis (e.g. rotenone or 2,4-dinitrophenol) is present in the incubation medium during exposure of the tissue to 125I-labelled rat IgG, it abolishes release of macromolecular radioactivity on re-incubation of the tissue. Enhanced tissue accumulation of 125I-labelled rat IgG, induced by the presence of leupeptin in the medium during the uptake phase, resulted in no concomitant increase in the amount of 125I-labelled IgG released in macromolecular form on re-incubation of the tissue. These findings indicate that the observed rapid release of 125I-labelled IgG is unlikely to represent release from lysosomes and is more compatible with release from a separate class of vesicle that does not fuse with lysosomes.
D019297:2,4-Dinitrophenol; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D004140:Dinitrophenols / Q000494:pharmacology; D005260:Female; D007074:Immunoglobulin G / Q000378:metabolism*; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D007976:Leupeptins / Q000494:pharmacology; D011205:Povidone / Q000494:pharmacology; D011247:Pregnancy; D051381:Rats; D012402:Rotenone / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000378:metabolism; D015017:Yolk Sac / Q000187:drug effects / Q000276:immunology*
D004140:Dinitrophenols; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D007976:Leupeptins; D000643:Ammonium Chloride; D012402:Rotenone; D012710:Serum Albumin, Bovine; D011205:Povidone; C032854:leupeptin; D019297:2,4-Dinitrophenol