Abstract 060: Angiotensin Type-2 Receptors In The Median Preoptic Nucleus Demarcate A Novel Depressor Neural Circuit In Mice

Abstract

Blood pressure (BP) is under constant control and regulation by an extensive neural network that spans a number of brain regions. This includes the median preoptic nucleus (MnPO) which directly projects to the pre-autonomic neurons of the hypothalamus to increase BP. While angiotensin type-1 receptors (AT 1 R) are expressed in the MnPO and project to the hypothalamus to increase BP, the role of angiotensin type-2 receptors (AT 2 R) is not clear. Here, we combine the use of AT 2 R-Cre mice, neuroanatomical techniques, in vivo optogenetics, pharmacology, and cardiovascular physiology to test the hypothesis that AT 2 R neurons of MnPO (MnPO AT2R ) elicit cardioprotective effects. We found an abundance of AT 2 R-containing neurons in the MnPO that expressed the mRNA(s) for AT 2 R with high fidelity (~77%; 241 of 313 neurons; n=7). Using RNAscope in situ hybridization, we found that ~ 42% (153 of 364 neurons; n=8) of MnPO AT2R expressed genetic markers for GABA, an inhibitory neurotransmitter. Next, we delivered a Cre-inducible AAV to direct expression of channelrhodopsin-2 (ChR2) and/or enhanced yellow fluorescent protein (eYFP) within MnPO AT2R of AT 2 R-Cre mice. Using in vitro electrophysiology, we confirmed functional expression of ChR2 within MnPO AT2R , with pulses of light reliably producing action potentials (n= 12 neurons/3 animals). Subsequently, we found that activation of MnPO AT2R elicited inhibitory post-synaptic currents in neighboring AT 2 R-negative neurons. The in vivo optogenetic excitation of MnPO AT2R elicited depressor responses in both adult male (ΔSBP: +2.8 ± 1.3 vs. -4.1 ± 1.9 mmHg, p<0.05 eYFP vs. ChR2, n=15) and female mice (ΔSBP: +0.8 ± 2.2 vs.-5.3 ± 1.9 mmHg, p<0.05 eYFP vs. ChR2, n=5). These responses were abolished in the presence of the autonomic ganglion blocker, hexamethonium. Finally, to assess whether the depressor effects were AT 2 R-mediated, we administered the selective AT 2 R-agonist, compound 21, into the MnPO and observed depressor responses (ΔSBP: +0.5 ± 0.3 vs.-4.9 ± 1.0 mmHg, p<0.05 Saline vs. C21, n=7). In conclusion, our results demonstrate that MnPO AT2R are depressor inhibitory neurons that can be targeted with compound 21 to lower blood pressure.