Radioautographic Visualization of the Incorporation of Galactose-3h and Mannose-3h by Rat Thyroids in Vitro in Relation to the Stages of Thyroglobulin Synthesis

Rat thyroid lobes incubated with mannose-(3)H, galactose-(3)H, or leucine-(3)H, were studied by radioautography. With leucine-(3)H and mannose-(3)H, the grain reaction observed in the light microscope is distributed diffusely over the cells at 5 min, with no reaction over the colloid. Later, the grains are concentrated towards the apex, and colloid reactions begin to appear by 2 hr. With galactose-(3)H, the reaction at 5 min is again restricted to the cells but it consists of clumped grains next to the nucleus. Soon after, grains are concentrated at the cell apex and colloid reactions appear in some follicles as early as 30 min. Puromycin almost totally inhibits incorporation of leucine-(3)H and mannose-(3)H, but has no detectable effect on galactose-(3)H incorporation during the 1st hr. Quantitation of electron microscope radioautographs shows that mannose-(3)H label localizes initially in the rough endoplasmic reticulum, and by 1-2 hr much of this reaction is transferred to the Golgi apparatus. At 3 hr and subsequently, significant reactions are present over apical vesicles and colloid, while the Golgi reaction declines. Label associated with galactose-(3)H localizes initially in the Golgi apparatus and rapidly transfers to the apical vesicles, and then to the colloid. These findings indicate that mannose incorporation into thyroglobulin precursors occurs within the rough endoplasmic reticulum; these precursors then migrate to the Golgi apparatus, where galactose incorporation takes place. The glycoprotein thus formed migrates via the apical vesicles to the colloid.