Experiments are described which test the hypothesis that the cuttlefish controls the relative volumes of gas space and liquid within its cuttlebone by an osmotic mechanism acting across the siphuncular surface of the bone. When the animal is at the bottom of the sea it would maintain the gas space within the cuttlebone by balancing the hydrostatic pressure of the sea by an osmotic force between the liquid within the cuttlebone and the blood.

In cuttlefish kept for some weeks in shallow water in an aquarium all the liquid taken from the cuttlebone is almost isotonic with the animals’ blood.

In animals recently hauled from the bottom of the sea the cuttlebone liquid is markedly hypotonic to sea water and hence to the blood.

These lower osmotic concentrations are given chiefly by reduction in the concentrations of the sodium and chloride ions.

After bringing animals up from about 70 m to the surface of the sea the osmotic concentration of the cuttlebone liquid rises from about 75% of sea water some 40 min after starting to haul the trawl, to about 97% of sea water 6 h later. Extrapolation back to the time at which hauling began gives a concentration of salts close to that predicted by the osmotic hypothesis.

Whilst the cuttlebone liquid is increasing in ionic concentration the liquid deeper in the cuttlebone is hypotonic to that just inside the siphuncular surface. This is explained in terms of the slowness of exchange of salts along the narrow channels between the lamellae of the cuttlebone.