The primary cilium functions as a mechanical and calcium signaling nexus

Cilia

Christopher R Jacobs⁴  Yingxiao Wang¹  Mardonn C Chua³  An M Nguyen²  Marie D Guevarra⁴  Kristen L Lee⁴ 

[1] Bioengineering Department, UC San Diego, La Jolla 92093, CA, USA;Jacobs Technion-Cornell Innovation Institute, Cornell Tech, New York 10011, NY, USA;Department of Biotechnology, University of British Columbia, Vancouver V6T 1Z4, BC, Canada;Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, MC 8904, 1210 Amsterdam Ave, New York 10027, NY, USA

关键词: Biosensor;    Kidney epithelia;    Osteocyte;    Calcium signaling;    Primary cilium;    Mechanotransduction;   

Others  :  1210200
DOI  :  10.1186/s13630-015-0016-y

 received in 2014-08-14, accepted in 2015-04-28,  发布年份 2015

【 摘 要 】

Background

The primary cilium is an antenna-like, nonmotile structure that extends from the surface of most mammalian cell types and is critical for chemosensing and mechanosensing in a variety of tissues including cartilage, bone, and kidney. Flow-induced intracellular calcium ion (Ca²⁺) increases in kidney epithelia depend on primary cilia and primary cilium-localized Ca²⁺-permeable channels polycystin-2 (PC2) and transient receptor potential vanilloid 4 (TRPV4). While primary cilia have been implicated in osteocyte mechanotransduction, the molecular mechanism that mediates this process is not fully understood. We directed a fluorescence resonance energy transfer (FRET)-based Ca²⁺ biosensor to the cilium by fusing the biosensor sequence to the sequence of the primary cilium-specific protein Arl13b. Using this tool, we investigated the role of several Ca²⁺-permeable channels that may mediate flow-induced Ca²⁺ entry: PC2, TRPV4, and PIEZO1.

Results

Here, we report the first measurements of Ca²⁺ signaling within osteocyte primary cilia using a FRET-based biosensor fused to ARL13B. We show that fluid flow induces Ca²⁺ increases in osteocyte primary cilia which depend on both intracellular Ca²⁺ release and extracellular Ca²⁺ entry. Using siRNA-mediated knockdowns, we demonstrate that TRPV4, but not PC2 or PIEZO1, mediates flow-induced ciliary Ca²⁺ increases and loading-induced Cox-2 mRNA increases, an osteogenic response.

Conclusions

In this study, we show that the primary cilium forms a Ca²⁺ microdomain dependent on Ca²⁺ entry through TRPV4. These results demonstrate that the mechanism of mechanotransduction mediated by primary cilia varies in different tissue contexts. Additionally, we anticipate that this work is a starting point for more studies investigating the role of TRPV4 in mechanotransduction.

【 授权许可】

   
2015 Lee et al.; licensee BioMed Central.

【 图 表 】

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