Antenatal betamethasone (BM) therapy accelerates lung development in preterm infants but may induce early programming events with long-term cardiovascular consequences. To elucidate these events, we developed a model of programming whereby pregnant ewes are administered BM (2 doses of 0.17 mg/kg) or vehicle at the 80th day of gestation and offspring are delivered at term. The BM-exposed (BMX) offspring develop elevated blood pressure, decreased baroreflex sensitivity, and alterations in the circulating, renal, and brain renin-angiotensin systems (RAS) by 6-months of age. We compared components of the choroid plexus 4th ventricle (ChP4) and CSF RAS between control and BMX male offspring at 6-months of age. In the ChP, high molecular weight renin protein and Ang I-intact angiotensinogen were unchanged between BMX and control animals. ACE2 activity was 3-fold higher than either NEP or ACE in control and BMX animals. Moreover, all three enzymes were equally enriched ~2.5 fold in ChP4 brush border membrane preparations. CSF Ang-(1-7) levels were significantly lower in BMX animals (351.8 ± 76.8 vs. 77.5 ± 29.7 fmol/mg; p<0.05) and ACE activity was significantly higher (6.6 ± 0.5 vs. 8.9 ± 0.5 fmol/min/ml; P<0.05) while ACE2 and NEP activities were below measurable limits. A thiol-sensitive peptidase contributed to the majority of Ang-(1-7) metabolism in the CSF, with higher activity in the BMX animals. We conclude that in utero BM exposure alters CSF but not ChP RAS components resulting in lower Ang-(1-7) levels in exposed animals.