Thyroid hormone effects on brain somatostatin-like immunoreactivity (SRIF-LI) were studied in an in vitro model system. Serum was removed from the nutrient culture medium of fetal day-18 rat cerebral cortex cells maintained in primary, long-term, dispersed monolayer culture. Chronic administration of either T3 or T4 in serum-free medium was associated with suppressed release of SRIF-LI into the culture medium (36-43 h accumulation), cell content of peptide and acute release in response to potassium-induced depolarization. Suppression was dose-dependent with an IC50 of less than 1 nM for T3. The most dramatic effects were observed for K+-induced release. Thirty-five to 50% suppression was typically observed with T3 at a near maximum dose (3 nM). Reverse T3 and diiodotyrosine were less potent and effective than T3. TRIAC and diiodothyronine also possessed significant suppressive activity. T3 suppression of release depended on duration of pretreatment. Administered for less than 16 h, T3 failed to significantly suppress K+-induced release, but significant suppression was observed for pretreatment periods of 16 h or longer. Indirect fluorescent immunohistochemical examination revealed a reduction in the number of cells positively stained for SRIF-LI in T3-treated dishes relative to controls. Upon removal of T3 and subsequent recovery in serum supplemented medium for 24 h, T3-treated and control cells exhibited similar levels of SRIF-LI release and cell content. T3-treated and control cells incorporated [3H] leucine into trichloracetic acid precipitable counts to similar extents. Dexamethasone and several sex steroids failed to modify the effects of T3 and did not independently influence SRIF-LI levels. Acute cycloheximide administration did not reverse T3 effects. The data indicate that primary brain cell cultures may be useful models to examine direct peripheral hormone actions on nervous tissue. Thyroid hormones suppress SRIF-LI levels in a dose, time and structure-dependent manner, which appears to be reversible. The findings are consistent with a possible integration of peripheral hormone and brain peptide physiology.