High glucose-induced apoptosis in cultured podocytes involves TRPC6-dependent calcium entry via the RhoA/ROCK pathway

He, Yang; Zhao, Bo; Liao, Chia-Shu; Zhang, Rui; Meng, Kexin; Xu, Jian; Jiao, Jundong

doi:10.1016/j.bbrc.2013.03.087

Cited by 56 publications

(51 citation statements)

References 26 publications

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“…TRPC6, a predominant member of these TRPC channels, plays a vital role in the regulation of cell functions, especially in dynamic actin cytoskeleton regulation [18, 19, 20]. Upon stimulation with angiotensin II or high glucose in podocytes, a TRPC6-mediated Ca 2+ influx increases the activity of RhoA, a small Rho GTPase, thereby leading to stress fiber formation to induce a stationary homeostatic podocyte phenotype [16, 21]. Interestingly, the renoprotective effects of CaSR agonists from cytoskeleton damage is associated with RhoA activity because treatment with the Rho-kinase inhibitor H1152, but not ERK1/2, p38 and JNK inhibitors, abolished R-568-induced increases in stress fiber and focal adhesion formation [9].…”

Section: Introductionmentioning

confidence: 99%

Calcium-Sensing Receptor Stimulation in Cultured Glomerular Podocytes Induces TRPC6-Dependent Calcium Entry and RhoA Activation

Zhang

Wang

et al. 2017

Cell Physiol Biochem

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Background/Aims: Recent studies provided compelling evidence that stimulation of the calcium sensing receptor (CaSR) exerts direct renoprotective action at the glomerular podocyte level. This protective action may be attributed to the RhoA-dependent stabilization of the actin cytoskeleton. However, the underlying mechanisms remain unclear. Methods: In the present study, an immortalized human podocyte cell line was used. Fluo-3 fluorescence was utilized to determine intracellular Ca²⁺ concentration ([Ca²⁺]_i), and western blotting was used to measure canonical transient receptor potential 6 (TRPC6) protein expression and RhoA activity. Stress fibers were detected by FITC-phalloidin. Results: Activating CaSR with a high extracellular Ca²⁺ concentration ([Ca²⁺]_o) or R-568 (a type II CaSR agonist) induces an increase in the [Ca²⁺]_i in a dose-dependent manner. This increase in [Ca²⁺]_i is phospholipase C (PLC)-dependent and is smaller in the absence of extracellular Ca²⁺ than in the presence of 0.5 mM [Ca²⁺]_o. The CaSR activation-induced [Ca²⁺]_i increase is attenuated by the pharmacological blockage of TRPC6 channels or siRNA targeting TRPC6. These data suggest that TRPC6 is involved in CaSR activation-induced Ca²⁺ influx. Consistent with a previous study, CaSR stimulation results in an increase in RhoA activity. However, the knockdown of TRPC6 significantly abolished the RhoA activity increase induced by CaSR stimulation, suggesting that TRPC6-dependent Ca²⁺ entry is required for RhoA activation. The activated RhoA is involved in the formation of stress fibers and focal adhesions in response to CaSR stimulation because siRNA targeting RhoA attenuated the increase in the stress fiber mediated by CaSR stimulation. Moreover, this effect of CaSR activation on the formation of stress fibers is also abolished by the knockdown of TRPC6. Conclusion: TRPC6 is involved in the regulation of stress fiber formation and focal adhesions via the RhoA pathway in response to CaSR activation. This may explain the direct protective action of CaSR agonists.

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Section: Introductionmentioning

confidence: 99%

Calcium-Sensing Receptor Stimulation in Cultured Glomerular Podocytes Induces TRPC6-Dependent Calcium Entry and RhoA Activation

Zhang

Wang

et al. 2017

Cell Physiol Biochem

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“…Others also showed that the activation of podocytes increases TRPC6 transcripts within 12 to 24 hours [25]. An increased TRPC6 expression may activate the calcium-calmodulin-dependent serine/threonine protein phosphatase, calcineurin, which dephosphorylates nuclear factor of activated T cells which then subsequently translocates to the cell nucleus.…”

Section: Discussionmentioning

confidence: 99%

Insulin Increases Expression of TRPC6 Channels in Podocytes by a Calcineurin-Dependent Pathway

Xia

Liu

et al. 2016

Cell Physiol Biochem

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Background/Aims: Insulin signaling to podocytes is relevant for the function of the glomerulus. Now, we tested the hypothesis that insulin increases the surface expression of canonical transient receptor potential canonical type 6 (TRPC6) channels in podocytes by a calcineurin-dependent pathway. Methods: We used quantitative RT-PCR, immunoblotting, immunofluorescence and fluorescence spectrophotometry in cultured podocytes. Activation of Nuclear Factor of Activated T-cells (NFATc1) was measured using a specific calorimetric assay. Results: Insulin increased the expression of TRPC6 transcripts and protein in podocytes. Insulin increased TRPC6 transcripts in a time and dose-dependent manner. The insulin-induced elevation of TRPC6 transcripts was blocked in the presence of tacrolimus, cyclosporine A, and NFAT-inhibitor (each p < 0.01 by ANOVA and Bonferroni's multiple comparison test). Transcripts of NOX4, another target gene of the calcineurin-NFAT pathway, were affected in a similar way. Immunoblotting showed that the administration of 100 nmol/L insulin increased TRPC6-proteins 2-fold within 48 hours. Insulin increased the activity of NFATc1 in nuclear extracts (p < 0.001) whereas tacrolimus, cyclosporine A, and NFAT-inhibitor blocked that insulin effect (p < 0.001; two way ANOVA). Immunofluorescence showed that insulin increased TRPC6-expression on the cell surface. Fluorescence-spectrophotometry and manganese quench experiments indicated that the increased TRPC6-expression after insulin administration was accompanied by an elevated transplasmamembrane cation influx. Insulin-stimulated surface expression of TRPC6 as well as transplasmamembrane cation influx could be reduced by pretreatment with tacrolimus. Conclusion: Insulin increases the expression of TRPC6 channels in podocytes by activation of the calcineurin-dependent pathway.

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“…Higher concentration of glucose increased the expression of TRPC6 and TRPC6-dependent Ca (2+) influx [22]. TRPC1 gene polymorphisms are associated with type 2 diabetes and diabetic nephropathy in Han Chinese population [23].…”

Section: Trpc1mentioning

confidence: 99%

Renal Membrane Transport Proteins and the Transporter Genes

Gowder¹

2014

Gene Technology

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High glucose-induced apoptosis in cultured podocytes involves TRPC6-dependent calcium entry via the RhoA/ROCK pathway

Cited by 56 publications

References 26 publications

Calcium-Sensing Receptor Stimulation in Cultured Glomerular Podocytes Induces TRPC6-Dependent Calcium Entry and RhoA Activation

Calcium-Sensing Receptor Stimulation in Cultured Glomerular Podocytes Induces TRPC6-Dependent Calcium Entry and RhoA Activation

Insulin Increases Expression of TRPC6 Channels in Podocytes by a Calcineurin-Dependent Pathway

Renal Membrane Transport Proteins and the Transporter Genes

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