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The contents and membranes of the secretory granules in the rat neurohypophysis were labelled in vivo with [³⁵S]cysteine and [³H]choline respectively. Density-gradient centrifugation of the labelled granules showed the membrane label to be distributed largely between two peaks: one associated with intact granules and one with the characteristics of empty granule envelopes. Stimulation of hormone release in vitro led to the movement of membrane label from the intact granule fraction to the other one, consistent with the recapture of membrane as large vesicles. Freezing and thawing the crude granule fraction, ostensibly to aid osmotic fracture, produced a single membrane component with a density intermediate between the two original fractions.