Lymphatic endothelial cell calcium pulses are sensitive to spatial gradients in wall shear stress

Cytosolic calcium (Ca2+) is a ubiquitous second messenger that influences numerous aspects of cellular function. In many cell types, cytosolic Ca2+ concentrations are characterized by periodic pulses, whose dynamics can influence downstream signal transduction. Here, we examine the general question of how cells use Ca2+ pulses to encode input stimuli in the context of the response of lymphatic endothelial cells (LECs) to fluid flow. Previous work shows that fluid flow regulates Ca2+ dynamics in LECs and that Ca2+-dependent signaling plays a key role in regulating lymphatic valve formation during embryonic development. However, how fluid flow might influence the Ca2+ pulse dynamics of individual LECs has remained, to our knowledge, little explored. We used live-cell imaging to characterize Ca2+ pulse dynamics in LECs exposed to fluid flow in an in vitro flow device that generates spatial gradients in wall shear stress (WSS), such as are found at sites of valve formation. We found that the frequency of Ca2+ pulses was sensitive to the magnitude of WSS, while the duration of individual Ca2+ pulses increased in the presence of spatial gradients in WSS. These observations provide an example of how cells can separately modulate Ca2+ pulse frequency and duration to encode distinct forms of information, a phenomenon that could extend to other cell types.

[1]  Hongkai Ji,et al.  Transcriptional outcomes and kinetic patterning of gene expression in response to NF-κB activation , 2018, PLoS biology.

[2]  M. Trebak,et al.  Pore properties of Orai1 calcium channel dimers and their activation by the STIM1 ER calcium sensor , 2018, The Journal of Biological Chemistry.

[3]  A. Orth,et al.  GPR68 Senses Flow and Is Essential for Vascular Physiology , 2018, Cell.

[4]  K. Red-Horse,et al.  DACH1 stimulates shear stress-guided endothelial cell migration and coronary artery growth through the CXCL12–CXCR4 signaling axis , 2017, Genes & development.

[5]  Y. Gwack,et al.  ORAI1 Activates Proliferation of Lymphatic Endothelial Cells in Response to Laminar Flow Through Krüppel-Like Factors 2 and 4 , 2017, Circulation research.

[6]  H. Vogel,et al.  Laminar flow downregulates Notch activity to promote lymphatic sprouting , 2017, The Journal of clinical investigation.

[7]  A. Dunn,et al.  Sphingosine 1-phosphate receptor 1 regulates the directional migration of lymphatic endothelial cells in response to fluid shear stress , 2016, Journal of The Royal Society Interface.

[8]  B. Cha,et al.  Mechanotransduction activates canonical Wnt/β-catenin signaling to promote lymphatic vascular patterning and the development of lymphatic and lymphovenous valves , 2016, Genes & development.

[9]  Ngan F Huang,et al.  Nanoscale Patterning of Extracellular Matrix Alters Endothelial Function under Shear Stress. , 2016, Nano letters.

[10]  K. Kawakami,et al.  Endothelial Ca2+ oscillations reflect VEGFR signaling-regulated angiogenic capacity in vivo , 2015, eLife.

[11]  D. Zawieja,et al.  Effects of dynamic shear and transmural pressure on wall shear stress sensitivity in collecting lymphatic vessels. , 2015, American journal of physiology. Regulatory, integrative and comparative physiology.

[12]  M. Delorenzi,et al.  FOXC2 and fluid shear stress stabilize postnatal lymphatic vasculature. , 2015, The Journal of clinical investigation.

[13]  Shin Lin,et al.  Novel mutations in PIEZO1 cause an autosomal recessive generalized lymphatic dysplasia with non-immune hydrops fetalis , 2015, Nature Communications.

[14]  P. Davies,et al.  Lymph flow regulates collecting lymphatic vessel maturation in vivo. , 2015, The Journal of clinical investigation.

[15]  Tyler D. Ross,et al.  Intramembrane binding of VE-cadherin to VEGFR2 and VEGFR3 assembles the endothelial mechanosensory complex , 2015, The Journal of cell biology.

[16]  R. Jahn,et al.  The GTPase Rab26 links synaptic vesicles to the autophagy pathway , 2015, eLife.

[17]  J. Moore,et al.  Measurement of shear stress-mediated intracellular calcium dynamics in human dermal lymphatic endothelial cells , 2015, American journal of physiology. Heart and circulatory physiology.

[18]  Tyler D Ross,et al.  Vascular remodeling is governed by a VEGFR3-dependent fluid shear stress set point , 2014, eLife.

[19]  D. Vittet,et al.  Lymphatic collecting vessel maturation and valve morphogenesis. , 2014, Microvascular research.

[20]  N. Yuldasheva,et al.  Piezo1 integration of vascular architecture with physiological force , 2014, Nature.

[21]  Shu Chien,et al.  Piezo1, a mechanically activated ion channel, is required for vascular development in mice , 2014, Proceedings of the National Academy of Sciences.

[22]  Erik Smedler,et al.  Frequency decoding of calcium oscillations. , 2014, Biochimica et biophysica acta.

[23]  Tom Verwijlen,et al.  Microvascular endothelial cells migrate upstream and align against the shear stress field created by impinging flow. , 2014, Biophysical journal.

[24]  E. Lammert,et al.  Mechanical forces in lymphatic vascular development and disease , 2013, Cellular and Molecular Life Sciences.

[25]  Nir Friedman,et al.  Dynamic response diversity of NFAT isoforms in individual living cells. , 2013, Molecular cell.

[26]  R. Adams,et al.  Mechanotransduction, PROX1, and FOXC2 cooperate to control connexin37 and calcineurin during lymphatic-valve formation. , 2012, Developmental cell.

[27]  D. Sheppard,et al.  Integrin-α9 is required for fibronectin matrix assembly during lymphatic valve morphogenesis , 2009, Developmental cell.

[28]  Junhao Hu,et al.  FOXC2 controls formation and maturation of lymphatic collecting vessels through cooperation with NFATc1 , 2009, The Journal of cell biology.

[29]  A. Akeson,et al.  NFATc1 regulates lymphatic endothelial development , 2009, Mechanisms of Development.

[30]  Jing Zhou,et al.  Endothelial Cilia Are Fluid Shear Sensors That Regulate Calcium Signaling and Nitric Oxide Production Through Polycystin-1 , 2008, Circulation.

[31]  D. Clapham,et al.  Calcium Signaling , 2007, Cell.

[32]  John A. Frangos,et al.  G protein-coupled receptors sense fluid shear stress in endothelial cells , 2006, Proceedings of the National Academy of Sciences.

[33]  Gerard L Cote,et al.  Lymph Flow, Shear Stress, and Lymphocyte Velocity in Rat Mesenteric Prenodal Lymphatics , 2006, Microcirculation.

[34]  J. Soboloff,et al.  Orai1 and STIM Reconstitute Store-operated Calcium Channel Function* , 2006, Journal of Biological Chemistry.

[35]  David A. Schultz,et al.  A mechanosensory complex that mediates the endothelial cell response to fluid shear stress , 2005, Nature.

[36]  G. Crabtree Calcium, Calcineurin, and the Control of Transcription* , 2001, The Journal of Biological Chemistry.

[37]  H. Kawakatsu,et al.  Shear Stress Stimulation of p130 cas Tyrosine Phosphorylation Requires Calcium-dependent c-Src Activation* , 1999, The Journal of Biological Chemistry.

[38]  Roger Y. Tsien,et al.  Cell-permeant caged InsP3 ester shows that Ca2+ spike frequency can optimize gene expression , 1998, Nature.

[39]  Keli Xu,et al.  Calcium oscillations increase the efficiency and specificity of gene expression , 1998, Nature.

[40]  Christopher C. Goodnow,et al.  Differential activation of transcription factors induced by Ca2+ response amplitude and duration , 1997, Nature.

[41]  R M Nerem,et al.  Pulsatile and steady flow-induced calcium oscillations in single cultured endothelial cells. , 1996, Journal of vascular research.

[42]  Bernd Nilius,et al.  Shear stress induced membrane currents and calcium transients in human vascular endothelial cells , 1992, Pflügers Archiv.

[43]  C F Dewey,et al.  Fluid shear stress modulates cytosolic free calcium in vascular endothelial cells. , 1992, The American journal of physiology.

[44]  J. Ando,et al.  Cytoplasmic calcium response to fluid shear stress in cultured vascular endothelial cells , 1988, In Vitro Cellular & Developmental Biology.

[45]  K. Miller Computational Biomechanics for Patient-Specific Applications , 2015, Annals of Biomedical Engineering.

[46]  O. F. Kampmeier,et al.  The origin and development of the venous valves, with particular reference to the saphenous district , 1927 .