Cockroach fat body incubated in a simple physiological saline solution releases trehalose to the surrounding medium. The output of trehalose occurs in the absence of ambient Ca2+ and decreases slowly with time. In two separate experiments, 0.1 mM CaCl2 added to the saline increased the output of trehalose on average by 70% but higher concentrations of Ca2+ did not further increase the efflux of trehalose. Stimulation of trehalose efflux by corpus cardiacum extract is absolutely dependent on extracellular Ca2+, no increase occurring beyond the basal level in the absence of the ion. The activity of corpus cardiacum extract increases as the concentration of CaCl2 is increased to 0.5 mM. This concentration of Ca2+ in the saline permits the extract to increase trehalose efflux by as much as 60% above the basal level. Corpus cardiacum extract, as well as the hypertrehalocaemic agents cyclic AMP and theophylline, increase significantly the influx of Ca2+ into fat body in vitro. The basal efflux of trehalose from fat body and that stimulated by corpus cardiacum extract is not dependent on extracellular Mg2+.