Angiotensin type 2 receptors in the intermediolateral cell column of the spinal cord: Negative regulation of sympathetic nerve activity and blood pressure

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Date of download: 5/28/2016 Copyright © The American College of Cardiology. All rights reserved. From: Contractile Function During Angiotensin-II Activation:

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Angiotensin type 2 receptors in the intermediolateral cell column of the spinal cord: Negative regulation of sympathetic nerve activity and blood pressure Jie Chao, Juan Gao, Karma-Jaya K. Parbhu, Lie Gao International Journal of Cardiology Volume 168, Issue 4, Pages 4046-4055 (October 2013) DOI: 10.1016/j.ijcard.2013.06.051 Copyright © 2013 The Authors Terms and Conditions

Fig. 1 Expression of AT1R and AT2R in spinal cord. A) Confocal immunofluorescent images showing AT1R and AT2R expressions in the T11 segment of the spinal cord. B) Western-blot showing AT1R and AT2R protein expressions in different regions of the spinal cord. (a) Representative blots and (b) mean data of blot density. #P<0.05 vs white matter, *P<0.05 vs IML; n=3 in each group. C) Gene expression of AT1R and AT2R in different regions of the spinal cord. All genes were examined by real-time RT-PCR with gene-specific primers. ΔC(t) (a) and relative fold change to IML (b) were calculated with CT value (see details in Methods). n=3 in each group. International Journal of Cardiology 2013 168, 4046-4055DOI: (10.1016/j.ijcard.2013.06.051) Copyright © 2013 The Authors Terms and Conditions

Fig. 2 Effect of Ang II on sympathetic nerve activity and blood pressure in the IML. A) Representative original recording of MAP and RSNA showing the responses to Ang II (a), Ang II following TEMPO (b), and Ang II following apocynin (c) in the IML. B) Mean data showing change of mean arterial pressure (a), sympathetic nerve activity (b), systolic pressure (c), diastolic pressure (d) and heart rate (e) after treatment with saline, Ang II, Ang II following TEMPO, or Ang II following apocynin, respectively. *P<0.05 vs saline group, #P<0.05 vs Ang II-treated group; n=5 in each group. International Journal of Cardiology 2013 168, 4046-4055DOI: (10.1016/j.ijcard.2013.06.051) Copyright © 2013 The Authors Terms and Conditions

Fig. 3 Effect of AT2R on sympathetic nerve activity and blood pressure. A) Representative original recording of MAP and RSNA showing the responses induced by CGP42112 (a), CGP42112 following PD123319 (b), and CGP42112 following l-NAME (c) in the IML. B) Mean data showing change of mean arterial pressure (a), sympathetic nerve activity (b), systolic pressure (c), diastolic pressure (d) and heart rate (e) after treatment of saline, CGP42112, CGP42112 following PD123319, or CGP42112 following l-NAME, respectively. *P<0.05 vs saline group, #P<0.05 vs Ang II-treated group; n=5 in each group. International Journal of Cardiology 2013 168, 4046-4055DOI: (10.1016/j.ijcard.2013.06.051) Copyright © 2013 The Authors Terms and Conditions

Fig. 4 AT2R activation inhibits the response to AT1R activation. A) Representative original recording of MAP and RSNA showing the responses induced by PD123319 (a), Ang II (b), and Ang II following PD123319 (c) in the IML. B) Mean data showing change of mean arterial pressure (a), sympathetic nerve activity (b), systolic pressure (c), diastolic pressure (d) and heart rate (e) after treatment of saline, PD123319, Ang II or Ang II following PD123319, respectively. *P<0.05 vs saline group, #P<0.05 vs Ang II-treated group; n=5 in each group. International Journal of Cardiology 2013 168, 4046-4055DOI: (10.1016/j.ijcard.2013.06.051) Copyright © 2013 The Authors Terms and Conditions

Fig. 5 Expression of AT1R and AT2R in acute isolated SPNs. Immunostaining of AT1R (A) or AT2R (B) is colocalized with MAP2 and ChAT-positive staining. AT1R and AT2R are colocalized in these neurons (C). DAPI is a nuclear marker; MAP2 is a mature neuronal marker; ChAT is the specific SPN marker. Scale bars, 20μm. International Journal of Cardiology 2013 168, 4046-4055DOI: (10.1016/j.ijcard.2013.06.051) Copyright © 2013 The Authors Terms and Conditions

Fig. 6 Effects of Ang II on electrophysiological characteristics of isolated IML neurons. A) Representative whole cell patch-clamp recording showing that the transient K+ current was enhanced by CGP42112 treatment (100 nM). B) Peak K+ current measured in response to a test pulse from −80 to +80mV. C) Complete current–voltage relationships from vehicle, CGP42112, PD123319, and PD123319+CGP42112 treatments. D) Resting membrane potential. Values are means±SE. *P<0.05 vs vehicle group. #P<0.05 vs CGP42112 group. n=10 in each group. International Journal of Cardiology 2013 168, 4046-4055DOI: (10.1016/j.ijcard.2013.06.051) Copyright © 2013 The Authors Terms and Conditions