Study of the mechanism of thyroid hormone secretion in an in vitro model system: indirect evidence for fusion of lysosomes with thyroglobulin liposomes.
The interaction between [131I]-thyroglobulin liposomes and thyroidal lysosomes was used as an in vitro model system for analyzing the relation of colloid droplets to lysosomes in follicular cells. The rates of hydrolysis of [131I]-thyroglobulin in a liposome-lysosome system (Lipo-Lyso system) and a thyroglobulin-lysosome system (TG-Lyso system) were compared. Hydrolysis of thyroglobulin in the Lipo-Lyso system increased hyperbolically and was greater than that in the TG-Lyso system for about 10 h. Liposomal thyroglobulin was not degraded by the 20,000 X g supernatant obtained on disruption of the lysosomal fraction. On varying the pH of the incubation medium, the highest activity was observed under acidic conditions in both systems. Under neutral or weakly alkaline conditions, the Lipo-Lyso system still showed 50% of the maximal hydrolytic activity, while the TG-Lyso system showed no activity. ATP and anaerobic conditions had no effect on either system. Cysteine (5 X 10-2M) and p-chloromercuribenzene sulfonic acid (PCMBS, 10-3 M) had not influence on hydrolysis in the Lipo-Lyso system, but in the TG-Lyso system cysteine greatly increased, and PCMBS significantly reduced the rate of hydrolysis. Dibutyryl cyclic AMP had no effect. Chlorpromazine (Cpz) decreased liposomal thyroglobulin hydrolysis in a concentration-dependent manner. In the TG-Lyso system, concentrations of 10-4M Cpz had no effect. These results strongly suggest that liposomes rapidly fused with lysosomes, providing optimal conditions for hydrolysis of thyroglobulin.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000693:Anaerobiosis; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D002746:Chlorpromazine; D003545:Cysteine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D066298:In Vitro Techniques; D008081:Liposomes / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D008954:Models, Biological; D051381:Rats; D013954:Thyroglobulin / Q000378:metabolism*; D013961:Thyroid Gland / Q000648:ultrastructure; D013963:Thyroid Hormones / Q000378:metabolism*; D013997:Time Factors
D002729:Chloromercuribenzoates; D008081:Liposomes; D013963:Thyroid Hormones; D003994:Bucladesine; D000255:Adenosine Triphosphate; D013954:Thyroglobulin; D003545:Cysteine; D002746:Chlorpromazine