Fractionation of iodinated particles and mitochondria from thyroid by zonal centrifugation and a study of their heterogeneity.
1. The subcellular particles of horse and rat thyroids were fractionated in a B XIV zonal rotor on a non-linear gradient of Ficoll after labelling with radioactive iodine in vitro (horse) or in vivo (rat). In the horse, the resulting fractions were analysed for radioactive iodine, protein and enzymes representative of certain subcellular particles. In the rat, iodine turnover and thyrotrophin stimulation were studied. 2. The population of iodinated particles could be subdivided into three main classes, characterized by differences in beta-galactosidase and acid phosphatase content and position in the gradient. The presence of a fourth class of particles is suggested. 3. It is concluded that iodinated particles isolated from the thyroid are essentially secondary lysosomes. Their heterogeneity is established with respect to their position in the gradient, their content of acid hydrolases and their iodine turnover. 4. The iodine pools of these secondary lysosomes are increased by thyrotrophin without any change in their number. 5. Their functional significance is discussed. 6. The distribution of mitochondria as judged by succinate dehydrogenase was also studied. The succinate dehydrogenase was spread throughout the gradient with a maximum of activity (40%) in the upper layer of the gradient. Separation of mitochondria from lysosomes by this method was not successful.
D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D002501:Centrifugation, Zonal; D005069:Evaluation Studies as Topic; D005696:Galactosidases / Q000032:analysis; D006736:Horses; D007457:Iodine Radioisotopes; D007467:Iodoproteins / Q000032:analysis; D007700:Kinetics; D008247:Lysosomes / Q000032:analysis; D008297:Male; D008722:Methods; D008928:Mitochondria / Q000032:analysis*; D051381:Rats; D013045:Species Specificity; D013053:Spectrophotometry; D013385:Succinate Dehydrogenase / Q000032:analysis; D013961:Thyroid Gland / Q000032:analysis / Q000166:cytology* / Q000201:enzymology
D007457:Iodine Radioisotopes; D007467:Iodoproteins; D013385:Succinate Dehydrogenase; D000135:Acid Phosphatase; D005696:Galactosidases