Effects of FSS on the expression and localization of the core proteins in two Wnt signaling pathways, and their association with ciliogenesis

Sheng,Xin; Sheng,Yan; Liu,Yuehua; Li,Xiaoqiong; Shu,Bo; Li,Dayu

doi:10.3892/ijmm.2018.3758

Effects of FSS on the expression and localization of the core proteins in two Wnt signaling pathways, and their association with ciliogenesis

Authors:
- Xin Sheng
- Yan Sheng
- Yuehua Liu
- Xiaoqiong Li
- Bo Shu
- Dayu Li
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Affiliations: Department of Biochemistry, Zunyi Medical University, Zunyi, Guizhou 563000, P.R. China, Laboratory of Basic Medical Morphology, Zunyi Medical University, Zunyi, Guizhou 563000, P.R. China
Published online on: July 5, 2018 https://doi.org/10.3892/ijmm.2018.3758
Pages: 1809-1818
Copyright: © Sheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Fluid shear stress (FSS) may alter ciliary structures and ciliogenesis, and it has been reported that the Wnt signaling pathway may regulate cilia assembly and disassembly. The present study aimed to investigate the effects of FSS on primary cilia, the Wnt/β‑catenin and Wnt/PCP signaling pathways, and the association among them. In the present study, human umbilical vein endothelial cells were subjected to FSS of differing velocities for various periods of time using a shear stress device. Subsequently, immunofluorescence and quantitative polymerase chain reaction were used to detect the expression and localization of the following core proteins: β‑catenin in the Wnt/β‑catenin signaling pathway; and dishevelled segment polarity protein 2 (Dvl2), fuzzy planar cell polarity protein (Fuz) and VANGL planar cell polarity protein 2 (Vangl2) in the Wnt/planar cell polarity (PCP) signaling pathway. Furthermore, the colocalization of Dvl2 with the basal body was analyzed under low FSS and laminar FSS. The results demonstrated that low FSS promoted the expression of Dvl2 and its colocalization with the basal body. Although Fuz expression was decreased with increasing duration of FSS, no visible alterations were detected in its localization, it was ubiquitously localized in the ciliated region. Conversely, the expression of Vangl2 was increased by laminar FSS, and β‑catenin was translocated into the nucleus at the early stage of low FSS. These findings suggested that Dvl2 may participate in low FSS‑induced ciliogenesis and β‑catenin may participate at the early stage, whereas Vangl2 may be associated with laminar FSS‑induced cilia disassembly.

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1	Hahn C and Schwartz MA: Mechanotransduction in vascular physiology and atherogenesis. Nat Rev Mol Cell Biol. 10:53–62. 2009. View Article : Google Scholar : PubMed/NCBI
2	Egorova AD, van der Heiden K, Poelmann RE and Hierck BP: Primary cilia as biomechanical sensors in regulating endothelial function. Differentiation. 83(Suppl): S56–S61. 2012. View Article : Google Scholar
3	Satir P, Pedersen LB and Christensen ST: The primary cilium at a glance. J Cell Sci. 123:499–503. 2010. View Article : Google Scholar : PubMed/NCBI
4	Shaheen R, Szymanska K, Basu B, Patel N, Ewida N, Faqeih E, Al Hashem A, Derar N, Alsharif H, Aldahmesh MA, et al: Characterizing the morbid genome of ciliopathies. Genome Biol. 17:2422016. View Article : Google Scholar : PubMed/NCBI
5	Nauli SM, Alenghat FJ, Luo Y, Williams E, Vassilev P, Li X, Elia AE, Lu W, Brown EM, Quinn SJ, et al: Polycystins 1 and 2 mediate mechanosensation in the primary cilium of kidney cells. Nat Genet. 33:129–137. 2003. View Article : Google Scholar : PubMed/NCBI
6	Nauli SM, Kawanabe Y, Kaminski JJ, Pearce WJ, Ingber DE and Zhou J: Endothelial cilia are fluid shear sensors that regulate calcium signaling and nitric oxide production through polycystin-1. Circulation. 117:1161–1171. 2008. View Article : Google Scholar : PubMed/NCBI
7	Marshall WF: Cilia self-organize in response to planar cell polarity and flow. Nat Cell Biol. 12:314–315. 2010. View Article : Google Scholar : PubMed/NCBI
8	Iomini C, Tejada K, Mo W, Vaananen H and Piperno G: Primary cilia of human endothelial cells disassemble under laminar shear stress. J Cell Biol. 164:811–817. 2004. View Article : Google Scholar : PubMed/NCBI
9	Van der Heiden K, Groenendijk BC, Hierck BP, Hogers B, Koerten HK, Mommaas AM, Gittenberger-de Groot AC and Poelmann RE: Monocilia on chicken embryonic endocardium in low shear stress areas. Dev Dyn. 235:19–28. 2006. View Article : Google Scholar
10	Van der Heiden K, Hierck BP, Krams R, de Crom R, Cheng C, Baiker M, Pourquie MJ, Alkemade FE, DeRuiter MC, Gittenberger-de Groot AC and Poelmann RE: Endothelial primary cilia in areas of disturbed flow are at the base of atherosclerosis. Atherosclerosis. 196:542–550. 2008. View Article : Google Scholar
11	Espinha LC, Hoey DA, Fernandes PR, Rodrigues HC and Jacobs CR: Oscillatory fluid flow influences primary cilia and microtubule mechanics. Cytoskeleton (Hoboken). 71:435–445. 2014. View Article : Google Scholar
12	Angers S and Moon RT: Proximal events in Wnt signal transduction. Nat Rev Mol Cell Biol. 10:468–477. 2009. View Article : Google Scholar : PubMed/NCBI
13	Maung SM and Jenny A: Planar cell polarity in Drosophila. Organogenesis. 7:165–179. 2011. View Article : Google Scholar : PubMed/NCBI
14	Boutin C, Labedan P, Dimidschstein J, Richard F, Cremer H, André P, Yang Y, Montcouquiol M, Goffinet AM and Tissir F: A dual role for planar cell polarity genes in ciliated cells. Proc Natl Acad Sci USA. 111:E3129–E3138. 2014. View Article : Google Scholar : PubMed/NCBI
15	Jia YY, Li F, Geng N, Gong P, Huang SJ, Meng LX, Lan J and Ban Y: Fluid flow modulates the expression of genes involved in the Wnt signaling pathway in osteoblasts in 3D culture conditions. Int J Mol Med. 33:1282–1288. 2014. View Article : Google Scholar : PubMed/NCBI
16	McCue S, Dajnowiec D, Xu F, Zhang M, Jackson MR and Langille BL: Shear stress regulates forward and reverse planar cell polarity of vascular endothelium in vivo and in vitro. Circ Res. 98:939–946. 2006. View Article : Google Scholar : PubMed/NCBI
17	Tatin F, Taddei A, Weston A, Fuchs E, Devenport D, Tissir F and Makinen T: Planar cell polarity protein Celsr1 regulates endothelial adherens junctions and directed cell rearrangements during valve morphogenesis. Dev Cell. 26:31–44. 2013. View Article : Google Scholar : PubMed/NCBI
18	Franco CA, Jones ML, Bernabeu MO, Vion AC, Barbacena P, Fan J, Mathivet T, Fonseca CG, Ragab A, Yamaguchi TP, et al: Non-canonical Wnt signalling modulates the endothelial shear stress flow sensor in vascular remodelling. Elife. 5:e077272016. View Article : Google Scholar : PubMed/NCBI
19	May-Simera HL and Kelley MW: Cilia, Wnt signaling, and the cytoskeleton. Cilia. 1:72012. View Article : Google Scholar
20	Wallingford JB and Mitchell B: Strange as it may seem: The many links between Wnt signaling, planar cell polarity, and cilia. Genes Dev. 25:201–213. 2011. View Article : Google Scholar : PubMed/NCBI
21	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
22	Bonewald LF and Johnson ML: Osteocytes, mechanosensing and Wnt signaling. Bone. 42:606–615. 2008. View Article : Google Scholar : PubMed/NCBI
23	Lara-Castillo N, Kim-Weroha NA, Kamel MA, Javaheri B, Ellies DL, Krumlauf RE, Thiagarajan G and Johnson ML: In vivo mechanical loading rapidly activates beta-catenin signaling in osteocytes through a prostaglandin mediated mechanism. Bone. 76:58–66. 2015. View Article : Google Scholar : PubMed/NCBI
24	Lai JK and Stainier DY: Pushing yap into the nucleus with shear force. Dev Cell. 40:517–518. 2017. View Article : Google Scholar : PubMed/NCBI
25	Sheng R, Kim H, Lee H, Xin Y, Chen Y, Tian W, Cui Y, Choi JC, Doh J, Han JK and Cho W: Cholesterol selectively activates canonical Wnt signalling over non-canonical Wnt signalling. Nat Commun. 5:43932014. View Article : Google Scholar : PubMed/NCBI
26	Chen X, Guo J, Yuan Y, Sun Z, Chen B, Tong X, Zhang L, Shen C and Zou J: Cyclic compression stimulates osteoblast differentiation via activation of the Wnt/beta-catenin signaling pathway. Mol Med Rep. 15:2890–2896. 2017. View Article : Google Scholar : PubMed/NCBI
27	Niu Q, Li F, Zhang L, Xu X, Liu Y, Gao J and Feng X: Role of the Wnt/beta-catenin signaling pathway in the response of chondrocytes to mechanical loading. Int J Mol Med. 37:755–762. 2016. View Article : Google Scholar : PubMed/NCBI
28	Curtis-Whitchurch L, Rekapally H and Hoying J: Phosphorylation and redistribution of Vangl2, a planar cell polarity protein, in endothelial cells in response to laminar shear stress. Faseb J. 31:2017.
29	Solter KM and Gibor A: The relationship between tonicity and flagellar length. Nature. 275:651–652. 1978. View Article : Google Scholar : PubMed/NCBI
30	Slough J, Cooney L and Brueckner M: Monocilia in the embryonic mouse heart suggest a direct role for cilia in cardiac morphogenesis. Dev Dyn. 237:2304–2314. 2008. View Article : Google Scholar : PubMed/NCBI
31	Li XM, Sheng X, Guo PF, Chen CH and He L: Changes on morphology and ciliogenesis of hUVECs loaded on different flow shear stress. Life Sci Res. 19:13–18. 2015.
32	Park TJ, Mitchell BJ, Abitua PB, Kintner C and Wallingford JB: Dishevelled controls apical docking and planar polarization of basal bodies in ciliated epithelial cells. Nat Genet. 40:871–879. 2008. View Article : Google Scholar : PubMed/NCBI
33	Lee KH, Johmura Y, Yu LR, Park JE, Gao Y, Bang JK, Zhou M, Veenstra TD, Yeon Kim B and Lee KS: Identification of a novel Wnt5a-CK1varepsilon-Dvl2-Plk1-mediated primary cilia disas-sembly pathway. EMBO J. 31:3104–3117. 2012. View Article : Google Scholar : PubMed/NCBI
34	Borovina A, Superina S, Voskas D and Ciruna B: Vangl2 directs the posterior tilting and asymmetric localization of motile primary cilia. Nat Cell Biol. 12:407–412. 2010. View Article : Google Scholar : PubMed/NCBI
35	Guirao B, Meunier A, Mortaud S, Aguilar A, Corsi JM, Strehl L, Hirota Y, Desoeuvre A, Boutin C, Han YG, et al: Coupling between hydrodynamic forces and planar cell polarity orients mammalian motile cilia. Nat Cell Biol. 12:341–350. 2010. View Article : Google Scholar : PubMed/NCBI
36	Fliegauf M, Benzing T and Omran H: When cilia go bad: Cilia defects and ciliopathies. Nat Rev Mol Cell Biol. 8:880–893. 2007. View Article : Google Scholar : PubMed/NCBI
37	Park TJ, Haigo SL and Wallingford JB: Ciliogenesis defects in embryos lacking inturned or fuzzy function are associated with failure of planar cell polarity and Hedgehog signaling. Nat Genet. 38:303–311. 2006. View Article : Google Scholar : PubMed/NCBI
38	Gray RS, Abitua PB, Wlodarczyk BJ, Szabo-Rogers HL, Blanchard O, Lee I, Weiss GS, Liu KJ, Marcotte EM, Wallingford JB and Finnell RH: The planar cell polarity effector Fuz is essential for targeted membrane trafficking, ciliogenesis and mouse embryonic development. Nat Cell Biol. 11:1225–1232. 2009. View Article : Google Scholar : PubMed/NCBI
39	Dai D, Zhu H, Wlodarczyk B, Zhang L, Li L, Li AG, Finnell RH, Roop DR and Chen J: Fuz controls the morphogenesis and differentiation of hair follicles through the formation of primary cilia. J Invest Dermatol. 131:302–310. 2011. View Article : Google Scholar
40	Zilber Y, Babayeva S, Seo JH, Liu JJ, Mootin S and Torban E: The PCP effector Fuzzy controls cilial assembly and signaling by recruiting Rab8 and Dishevelled to the primary cilium. Mol Biol Cell. 24:555–565. 2013. View Article : Google Scholar : PubMed/NCBI
41	Brooks ER and Wallingford JB: Control of vertebrate intraflagellar transport by the planar cell polarity effector Fuz. J Cell Biol. 198:37–45. 2012. View Article : Google Scholar : PubMed/NCBI
42	Lancaster MA, Schroth J and Gleeson JG: Subcellular spatial regulation of canonical Wnt signalling at the primary cilium. Nat Cell Biol. 13:700–707. 2011. View Article : Google Scholar : PubMed/NCBI