Shear stress-induced Ca2+ mobilization in MDCK cells is ATP dependent, no matter the primary cilium

Cell Calcium ◽  
2013 ◽  
Vol 53 (5-6) ◽  
pp. 327-337 ◽  
Author(s):  
Lise Rodat-Despoix ◽  
Jizhe Hao ◽  
Mathieu Dandonneau ◽  
Patrick Delmas
Keyword(s):  
Shear Stress ◽  
Primary Cilium ◽  
Mdck Cells

Reduction of oxidative stress during recovery accelerates normalization of primary cilia length that is altered after ischemic injury in murine kidneys

2013 ◽  
Vol 304 (10) ◽  
pp. F1283-F1294 ◽  
Author(s):  
Jee In Kim ◽  
Jinu Kim ◽  
Hee-Seong Jang ◽  
Mi Ra Noh ◽  
Joshua H. Lipschutz ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Primary Cilium ◽  
Primary Cilia ◽  
Mdck Cells ◽  
Reactive Oxygen ◽  
Renal Tubular Cell ◽  
Oxygen Species ◽  
Renal Tubular ◽  
Cilium Length

The primary cilium is a microtubule-based nonmotile organelle that extends from the surface of cells, including renal tubular cells. Here, we investigated the alteration of primary cilium length during epithelial cell injury and repair, following ischemia/reperfusion (I/R) insult, and the role of reactive oxygen species in this alteration. Thirty minutes of bilateral renal ischemia induced severe renal tubular cell damage and an increase of plasma creatinine (PCr) concentration. Between 8 and 16 days following the ischemia, the increased PCr returned to normal range, although without complete histological restoration. Compared with the primary cilium length in normal kidney tubule cells, the length was shortened 4 h and 1 day following ischemia, increased over normal 8 days after ischemia, and then returned to near normal 16 days following ischemia. In the urine of I/R-subjected mice, acetylated tubulin was detected. The cilium length of proliferating cells was shorter than that in nonproliferating cells. Mature cells had shorter cilia than differentiating cells. Treatment with Mn(III) tetrakis(1-methyl-4-pyridyl) porphyrin (MnTMPyP), an antioxidant, during the recovery of damaged kidneys accelerated normalization of cilia length concomitant with a decrease of oxidative stress and morphological recovery in the kidney. In the Madin-Darby canine kidney (MDCK) cells, H2O2 treatment caused released ciliary fragment into medium, and MnTMPyP inhibited the deciliation. The ERK inhibitor U0126 inhibited elongation of cilia in normal and MDCK cells recovering from H2O2 stress. Taken together, our results suggest that primary cilia length reflects cell proliferation and the length of primary cilium is regulated, at least, in part, by reactive oxygen species through ERK.

Shear stress turns on the primary cilium and lipophagy

2020 ◽  
Vol 22 (9) ◽  
pp. 1029-1030
Author(s):  
Nuria Martinez-Lopez ◽  
Rajat Singh
Keyword(s):  
Shear Stress ◽  
Primary Cilium

Transepithelial pressure pulses induce nucleotide release in polarized MDCK cells

2005 ◽  
Vol 288 (1) ◽  
pp. F133-F141 ◽  
Author(s):  
H. A. Praetorius ◽  
J. Frøkiær ◽  
J. Leipziger
Keyword(s):  
Epithelial Cells ◽  
Laminar Flow ◽  
Primary Cilium ◽  
Mdck Cells ◽  
Chamber Pressure ◽  
Dependent Manner ◽  
Nucleotide Release ◽  
Pressure Pulses ◽  
Kidney Epithelial Cells ◽  
Pressure Changes

The release of nucleotides is involved in mechanosensation in various epithelial cells. Intriguingly, kidney epithelial cells are absolutely dependent on the primary cilium to sense changes in apical laminar flow. During fluid passage, the renal epithelial cells are subjected to various mechanical stimuli in addition to changes in the laminar flow rate. In the distal part of the collecting duct, the epithelial cells are exposed to pressure changes and possibly distension during papillary contractions. The aim of the present study was to determine whether nucleotide release contributes to mechanosensation in kidney epithelial cells, thereby establishing whether pressure changes are sufficient to produce nucleotide-mediated responses. Madin-Darby canine kidney (MDCK) cells grown on permeable supports were mounted in a closed double perfusion chamber on an inverted microscope. The intracellular Ca2+ concentration ([Ca2+]i) was monitored with the Ca2+-sensitive fluorescence probe fluo 4. Transepithelial pressure pulses of 30–80 mmHg produced a transient increase in [Ca2+]i of MDCK cells. This response is independent of the primary cilium, since it is readily observed in immature cells that do not yet express primary cilia. The amplitudes of the pressure-induced Ca2+ transients varied with the applied chamber pressure in a quantity-dependent manner. The ATPase apyrase and the P2Y antagonist suramin significantly reduced the pressure-induced Ca2+ transients. Applying apyrase or suramin to both sides of the preparation simultaneously nearly abolished the pressure-induced Ca2+ response. In conclusion, these observations suggest that rapid pressure changes induce both apical and basolateral nucleotide release that contribute to mechanosensation in kidney epithelial cells.

scholarly journals PI3KC2α-dependent and VPS34-independent generation of PI3P controls primary cilium-mediated autophagy in response to shear stress

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Asma Boukhalfa ◽  
Anna Chiara Nascimbeni ◽  
Damien Ramel ◽  
Nicolas Dupont ◽  
Emilio Hirsch ◽  
...  
Keyword(s):  
Shear Stress ◽  
Primary Cilium

β1-Integrins in the primary cilium of MDCK cells potentiate fibronectin-induced Ca2+ signaling

2004 ◽  
Vol 287 (5) ◽  
pp. F969-F978 ◽  
Author(s):  
H. A. Praetorius ◽  
J. Praetorius ◽  
S. Nielsen ◽  
J. Frokiaer ◽  
K. R. Spring
Keyword(s):  
Primary Cilium ◽  
Primary Cilia ◽  
Mdck Cells ◽  
Immunogold Electron Microscopy ◽  
Induced Response ◽  
Collecting Ducts ◽  
Intracellular Ca ◽  
Induced Signal ◽  
Integrin Ligand ◽  
Darby Canine Kidney

Because β1-integrin is involved in sensing of fluid flow rate in endothelial cells, a function that in Madin-Darby canine kidney (MDCK) cells is confined to the primary cilium, we hypothesized β1-integrin to be an important part of the primary ciliary mechanosensory apparatus in MDCK cells. We observed that β1-integrin, α3-integrin, and perhaps α5-integrin were localized to the primary cilium of MDCK cells by combining lectin and immunofluorescence confocal microscopy. β1-Integrin was also colocalized with tubulin to the primary cilia of the rat renal collecting ducts, as well as to the cilia of proximal tubules and thick ascending limbs. Immunogold-electron microscopy confirmed the presence of β1-integrin on primary cilia of MDCK cells and rat collecting ducts. Intracellular Ca2+ levels, monitored by fluorescence microscopy on fluo 4-loaded MDCK cells, significantly increased on addition of fibronectin, a β1-integrin ligand, to mature MDCK cells with an IC50 of 0.02 mg/l. In immature, nonciliated cells or in deciliated mature cells, the IC50 was 0.40 mg/l. Blocking the fibronectin-binding sites of β1-integrin with RGD peptide prevented the Ca2+ signal. Cross-linking of β1-integrins by Sambucus nigra agglutinin produced a Ca2+ response similar to the addition of fibronectin. Furthermore, the fibronectin-induced response was not dependent on flow or a flow-induced Ca2+ response. Finally, the flow-induced Ca2+ response was not prevented by the fibronectin-induced signal. Although β1-integrin on the primary cilium greatly potentiates the fibronectin-induced Ca2+ signaling in MDCK cells, the flow-dependent Ca2+ signal is not mediated through activation of β1-integrin.

Bending the Primary Cilium Opens Ca2+-sensitive Intermediate-Conductance K+ Channels in MDCK Cells

2003 ◽  
Vol 191 (3) ◽  
pp. 193-200 ◽  
Author(s):  
H.A. Praetorius ◽  
J. Frokiaer ◽  
S. Nielsen ◽  
K.R. Spring
Keyword(s):  
Primary Cilium ◽  
Mdck Cells ◽  
K Channels

scholarly journals Primary Cilia and Atherosclerosis

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhi-Mei Wang ◽  
Xiao-Fei Gao ◽  
Jun-Jie Zhang ◽  
Shao-Liang Chen
Keyword(s):  
Blood Flow ◽  
Shear Stress ◽  
Cardiovascular System ◽  
Primary Cilium ◽  
Primary Cilia ◽  
Laminar Shear Stress ◽  
Atrioventricular Canal ◽  
Intracellular Space ◽  
Mesenchymal Transition ◽  
Laminar Shear

In artery tree, endothelial function correlates with the distribution of shear stress, a dragging force generated by flowing blood. In laminar shear stress areas, endothelial cells (ECs) are available to prevent atherosclerosis, however, ECs in disturbed shear stress sites are featured with proinflammation and atherogenesis. Basic studies in the shear stress field that focused on the mechanosensors of ECs have attracted the interest of researchers. Among all the known mechanosensors, the primary cilium is distinctive because it is enriched in disturbed shear stress regions and sparse in laminar shear stress areas. The primary cilium, a rod liked micro-organelle, can transmit extracellular mechanical and chemical stimuli into intracellular space. In the cardiovascular system, primary cilia are enriched in disturbed shear stress regions, where blood flow is slow and oscillatory, such as the atrium, downstream of the aortic valve, branches, bifurcations, and inner curves of the artery. However, in the atrioventricular canal and straight vessels, blood flow is laminar, and primary cilia can barely be detected. Primary cilia in the heart cavity prevent ECs from mesenchymal transition and calcification by suppressing transforming growth factor (TGF) signaling. Besides, primary cilia in the vascular endothelium protected ECs against disturbed shear stress-induced cellular damage by triggering Ca2+ influx as well as nitric oxide (NO) release. Moreover, primary cilia inhibit the process of atherosclerosis. In the current review, we discussed ciliogenesis, ciliary structure, as well as ciliary distribution, function and the coordinate signal transduction with shear stress in the cardiovascular system.

scholarly journals Flow‐induced shear stress reduces K content of MDCK cells by a Ca and TEA‐dependent K efflux mechanism

2008 ◽  
Vol 22 (S1) ◽  
Author(s):  
J. David Holtzclaw ◽  
P. Richard Grimm ◽  
Debra L. Irsik ◽  
Steven C. Sansom
Keyword(s):  
Shear Stress ◽  
Mdck Cells ◽  
Efflux Mechanism

scholarly journals The ciliary membrane of polarized epithelial cells stems from a midbody remnant-associated membrane patch with condensed nanodomains

2019 ◽  
Author(s):  
Miguel Bernabé-Rubio ◽  
Minerva Bosch-Fortea ◽  
Esther García ◽  
Jorge Bernardino de la Serna ◽  
Miguel A. Alonso
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Primary Cilium ◽  
Mdck Cells ◽  
Cell Types ◽  
Apical Surface ◽  
Cellular Behavior ◽  
Ciliary Membrane ◽  
Lipid Dynamics

AbstractThe primary cilium is a specialized plasma membrane protrusion that harbors receptors involved in important signaling pathways. Despite its central role in regulating cellular behavior, the biogenesis of the primary cilium is not fully understood. In fact, the source of the ciliary membrane remains a mystery in cell types that assemble their primary cilium entirely at the cell surface, such as polarized renal epithelial cells. After cytokinesis, the remnant of the midbody of these cells moves to the center of the apical surface, where it licenses the centrosome for ciliogenesis through an unidentified mechanism. Here, to investigate the origin of the ciliary membrane and the role of the midbody remnant, we analyzed membrane compaction and lipid dynamics at the microscale and nanoscale in living renal epithelial MDCK cells. We found that a specialized patch made of condensed membranes with restricted lipid lateral mobility surrounds the midbody remnant. This patch accompanies the remnant on its journey towards the centrosome and, once the two structures have met, the remnant delivers part of membranes of the patch to build the ciliary membrane. In this way, we have determined the origin of the ciliary membrane and the contribution of the midbody remnant to primary cilium formation in cells whose primary cilium is assembled at the plasma membrane.

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