1 An organ culture system is described for the in vitro maintenance of superior cervical sympathetic ganglia taken from mice of any age. The relation of tyrosine hydroxylase (T-OH) activity to ganglionic noradrenaline (NA) content has been investigated under various culture conditions.2 Depolarizing stimuli such as raised extracellular potassium and ouabain evoked increases of approximately 70% in the T-OH activity of cultured ganglia over a 48 h period. Exposure to a high concentration of potassium (high K(+)) for 30 min at the start of a 48 h culture was sufficient to elicit significant increases in T-OH activity.3 Depolarization-induced rises in T-OH activity were observed after culture in the presence or absence of nerve growth factor.4 The NA content of ganglia, cultured for 48 h in the presence of high K(+), ouabain, reserpine, clorgyline and alpha-methyl-p-tyrosine, showed no constant relation to their T-OH activity.5 Dibutyryl cyclic adenosine 3'-5'-monophosphate (dibutyryl cyclic AMP) mimicked high K(+) in its effect on ganglionic T-OH activity and NA content. Theophylline enhanced the potassium effects.6 Rises in the T-OH activity of ganglia cultured in the presence of high K(+) and dibutyryl cyclic AMP were abolished if the protein synthesis inhibitors cycloheximide or actinomycin D were present in the culture medium.7 It is concluded that the link between prolonged depolarization and rises in T-OH activity does not seem to depend upon changes in ganglionic NA content. In the intact animals, trans-synaptic modulation may take the form of a depolarization-induced rise in the cyclic AMP content of sympathetic ganglionic neurones leading to nuclear mediated synthesis of T-OH.