DELIVERY OF ANGIOTENSIN-II TYPE-1 RECEPTOR ANTISENSE INHIBITS ANGIOTENSIN ACTION IN NEURONS FROM HYPERTENSIVE RAT-BRAIN

Increased brain angiotensin II (AII) type 1 receptor (AT(1)R) expression has been implicated in the hyperactive brain angiotensin system and the development and maintenance of hypertension in the genetically spontaneously hypertensive (SH) rat. Neuronal cells in primary culture from the cardioregulatory-relevant brain areas (hypothalamus/brainstem) mimic increased brain AT(1)R gene expression and AT(1)R function of the adult SH rat. They have been utilized in the present study to determine whether cellular actions of AII could be regulated by the transfer of AT(1)R antisense (AT(1)R.AS) with the use of a retroviral-mediated gene delivery system developed for the central nervous system cultures. AII stimulates norepinephrine (NE) uptake in neuronal cultures of both normotensive (Wistar Kyoto) and SH rat brains. This neuromodulatory action is mediated by the AT(1)R subtype, is significantly higher in SH neurons, and is associated with a parallel stimulation of mRNAs for c-fos and NE transporter. Infection of neuronal cultures with a retrovirus vector that contains AT(1)R-AS (LNSV-AT(1)R-AS) results in an inhibition of AT(1)R-mediated stimulation of both c-fos and NE transporter mRNA, as well as NE uptake in both strains of rats; however, the inhibition is more pronounced in SH neurons compared with Wistar Kyoto rat brain neurons. The higher sensitivity of the SH rat brain neurons is further supported by our observation that a certain dose of LNSV-AT(1)R-AS that fails to induce inhibition of cellular actions of AII in WKY neurons causes a significant inhibition of AII actions in SH neurons. These observations show that retrovirally mediated delivery of AT(1)R-AS could be used to selectively control the actions of AII in primary neuronal cultures from SH rat brain.