A time sequence study was performed on experimental animals to investigate long-term effects of intravenously administered glycerol on the epithelial cell activity in the endolymphatic sac (ES) and on the ES volume. Fifteen to 60 min after systemic glycerol administration, the ES volume decreased. During this time, the ES lumen was often obliterated. Subsequently, the lumen dilated. Meanwhile, many light epithelial cells showed granules with floccular and/or lamellar contents. Concomitant deposition of floccular material into the luminal space suggested secretion of macromolecular substances, presumably from these transformed light cells. The number of granule-containing cells was significantly increased 2 h (p < 0.01) and 4 h (p < 0.01) after glycerol administration. The ES was significantly dilated after 4 h (p < 0.01) and 6 h (p < 0.05). Thus systemic alterations in osmotic pressure led to a reversible change in ES volume, with initial collapse followed by dilation and normalization after 8 h. The secretory response of the ES preceded the volume increase. A great variability in ES volume indicated high compliance of this organ system. A secretion/degradation system or turnover of osmotically active macromolecular complexes in the epithelial lining and ES lumen seems to be linked to the ability of the ES to hold fluid volumes within a wide range. This may serve as a micromechanical pressure-volume-regulating device for monitoring endolymph fluid homeostasis.