TY - JOUR
T1 - The role of angiotensin II in glomerular volume dynamics and podocyte calcium handling
AU - Ilatovskaya, Daria V.
AU - Palygin, Oleg
AU - Levchenko, Vladislav
AU - Endres, Bradley T.
AU - Staruschenko, Alexander
N1 - Funding Information:
The authors would like to thank Colleen Lavin (Nikon Inc.) for the technical advice. Drs Yuri A. Negulyaev and Vladislav Chubinskiy-Nadezhdin (Institute of Cytology RAS, St. Petersburg, Russia) are acknowledged for the assistance with the initial calcium imaging experiments. The authors are grateful to Ruben Sandoval (Indiana University) for the technical advice with fluorescent dextrans. This research was partially supported by the NIH R01 HL108880, R01 HL122662, R35 HL135749 (to AS), and K99 DK105160 (to DVI), ADA grant 1–15-BS-172 (to AS), ASN Ben J. Lipps fellowship (to DVI), AHA 16EIA26720006 (to AS), JDRF 1-INO-2016–223-A-N (to AS) and NKF Young Investigator Grant (to OP). Sanofi-Aventis Deutschland GmbH is acknowledged for providing SAR7334.
Publisher Copyright:
© The Author(s) 2017.
PY - 2017
Y1 - 2017
N2 - Podocytes are becoming a primary focus of research efforts due to their association with progressive glomeruli damage in disease states. Loss of podocytes can occur as a result of excessive intracellular calcium influx, and we have previously shown that angiotensin II (Ang II) via canonical transient receptor potential 6 (TRPC6) channels caused increased intracellular Ca2+ flux in podocytes. We showed here with patch-clamp electrophysiology that Ang II activates TRPC channels; then using confocal calcium imaging we demonstrated that Ang II–dependent stimulation of Ca2+ influx in the podocytes is precluded by blocking either AT1 or AT2 receptors (ATRs). Application of Ang(1–7) had no effect on intracellular calcium. Ang II-induced calcium flux was decreased upon inhibition of TRPC channels with SAR7334, SKF 96365, clemizole hydrochloride and La3+, but not ML204. Using a novel 3D whole-glomerulus imaging ex vivo assay, we revealed the involvement of both ATRs in controlling glomerular permeability; additionally, using specific inhibitors and activators of TRPC6, we showed that these channels are implicated in the regulation of glomerular volume dynamics. Therefore, we provide evidence demonstrating the critical role of Ang II/TRPC6 axis in the control of glomeruli function, which is likely important for the development of glomerular diseases.
AB - Podocytes are becoming a primary focus of research efforts due to their association with progressive glomeruli damage in disease states. Loss of podocytes can occur as a result of excessive intracellular calcium influx, and we have previously shown that angiotensin II (Ang II) via canonical transient receptor potential 6 (TRPC6) channels caused increased intracellular Ca2+ flux in podocytes. We showed here with patch-clamp electrophysiology that Ang II activates TRPC channels; then using confocal calcium imaging we demonstrated that Ang II–dependent stimulation of Ca2+ influx in the podocytes is precluded by blocking either AT1 or AT2 receptors (ATRs). Application of Ang(1–7) had no effect on intracellular calcium. Ang II-induced calcium flux was decreased upon inhibition of TRPC channels with SAR7334, SKF 96365, clemizole hydrochloride and La3+, but not ML204. Using a novel 3D whole-glomerulus imaging ex vivo assay, we revealed the involvement of both ATRs in controlling glomerular permeability; additionally, using specific inhibitors and activators of TRPC6, we showed that these channels are implicated in the regulation of glomerular volume dynamics. Therefore, we provide evidence demonstrating the critical role of Ang II/TRPC6 axis in the control of glomeruli function, which is likely important for the development of glomerular diseases.
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U2 - 10.1038/s41598-017-00406-2
DO - 10.1038/s41598-017-00406-2
M3 - Article
C2 - 28331185
AN - SCOPUS:85026782870
SN - 2045-2322
VL - 7
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 299
ER -