Most of our knowledge of the function of the angiotensin type 2 receptor (AT 2 R) has been obtained from transgenic mouse models. The aim of the present study was to investigate the role of the AT 2 R in normotensive Sprague-Dawley (SD) rats by using antisense gene transfer technology to 'knockdown' this specific receptor subtype. A retroviral vector containing full-length AT 2 R antisense cDNA (AT 2 R-AS) was constructed and the effectiveness of the transduction of AT 2 R-AS was studied in vitro. In subsequent in vivo studies, 5-day-old normotensive SD rats received a single intracardiac bolus (25 µl) of AT 2 R-AS viral particles. When animals reached adulthood, direct blood pressure (BP), and both pressor and dipsogenic responses to angiotensin II were investigated. Long-lasting expression of the AT 2 R-AS transcript and a reduction in mRNA and binding of the AT 2 R was observed in vitro. Expression of AT 2 R-AS transcript was maintained for 90 days in heart, kidney, lung and brain, indicating a high degree of transgene transduction in vivo. As adults, systolic BP and the pressor responses to angiotensin were significantly elevated in AT 2 R-AS-treated rats. However, AT 2 R-AS-treated rats displayed significantly reduced dipsogenic responses to both angiotensin and water deprivation. Collectively, these data demonstrate that a single neonatal injection of the retroviral vector containing antisense to the AT 2 receptors in rats results in similar cardiovascular and dipsogenic responses as reported in AT 2 R knockout mice.