Functional characterization of TRPV4 as an osmotically sensitive ion channel in porcine articular chondrocytes.
Functional characterization of TRPV4 as an osmotically sensitive ion channel in porcine articular chondrocytes.
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DOI：
10.1002/art.24799
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发表时间：
2009-10
影响因子：
--
通讯作者：
Guilak, Farshid
中科院分区：
文献类型：
--
作者：
Phan, Mimi N.;Leddy, Holly A.;Votta, Bartholomew J.;Kumar, Sanjay;Levy, Dana S.;Lipshutz, David B.;Lee, Suk Hee;Liedtke, Wolfgang;Guilak, Farshid
瞬时受体电位香草蛋白4 （TRPV4）是Ca2+可渗透通道，可通过强直性（渗透压）和机械刺激进行门控。软骨细胞，软骨中的细胞，对它们的渗透和机械环境作出反应；然而，这种信号转导的分子基础尚不完全清楚。本研究的目的是证明TRPV4在软骨细胞中的存在和功能。免疫标记法和Western blotting法检测TRPV4蛋白表达。为了响应TRPV4激动剂/拮抗剂、渗透应激和白细胞介素-1 (IL-1)，分别使用荧光显微镜、光学显微镜或免疫测定法测量了猪软骨细胞中Ca2+信号、细胞体积和前列腺素E2 （PGE2）产生的变化。TRPV4在RNA和蛋白水平上大量表达。暴露于4αPDD（一种TRPV4激活剂）下，导致软骨细胞中的Ca2+信号传导，该信号被选择性TRPV4拮抗剂GSK205阻断。阻断TRPV4降低了软骨细胞对低渗透应激的反应，减少了Ca2+反应细胞的比例、调节体积减少（RVD）和PGE2的产生。Ca2+信号被细胞外Ca2+的去除或细胞内储存的消耗所抑制。特异性激活TRPV4可恢复IL-1引起的RVD缺陷。化学破坏初级纤毛消除Ca2+信号，以响应4αPDD或低渗透胁迫。TRPV4存在于关节软骨细胞中，软骨细胞对低渗透应激的反应是由该通道介导的，该通道涉及细胞外Ca2+和细胞内Ca2+释放。TRPV4也可能参与调节促炎分子的产生或对渗透应激的影响。
Transient receptor potential vanilloid 4 (TRPV4) is a Ca2+ permeable channel that can be gated by tonicity (osmolarity) and mechanical stimuli. Chondrocytes, the cells in cartilage, respond to their osmotic and mechanical environments; however, the molecular basis of this signal transduction is not fully understood. The objective of this study was to demonstrate the presence and functionality of TRPV4 in chondrocytes. TRPV4 protein expression was measured by immunolabeling and Western blotting. In response to TRPV4 agonist/antagonists, osmotic stress, and interleukin-1 (IL-1), changes in Ca2+ signaling, cell volume, and prostaglandin E2 (PGE2) production were measured in porcine chondrocytes using fluorescence microscopy, light microscopy, or immunoassay, respectively. TRPV4 was expressed abundantly at the RNA and protein level. Exposure to 4αPDD, a TRPV4 activator, caused Ca2+ signaling in chondrocytes, which was blocked by the selective TRPV4 antagonist, GSK205. Blocking TRPV4 diminished the chondrocytes' response to hypo-osmotic stress, reducing the fraction of Ca2+ responsive cells, regulatory volume decrease (RVD), and PGE2 production. Ca2+ signaling was inhibited by removal of extracellular Ca2+ or depletion of intracellular stores. Specific activation of TRPV4 restored defective RVD caused by IL-1. Chemical disruption of the primary cilium eliminated Ca2+ signaling in response to either 4αPDD or hypo-osmotic stress. TRPV4 is present in articular chondrocytes, and chondrocyte response to hypo-osmotic stress is mediated by this channel, which involves both an extracellular Ca2+ and intracellular Ca2+ release. TRPV4 may also be involved in modulating the production or influence of pro-inflammatory molecules in response to osmotic stress.
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影响因子：
4.4
作者：
Jefferies, D;Farquharson, C;Whitehead, C
通讯作者：
Whitehead, C
DOI：
10.1083/jcb.93.3.743
发表时间：
1982-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kuettner KE;Pauli BU;Gall G;Memoli VA;Schenk RK
通讯作者：
Schenk RK
影响因子：
3.6
作者：
Chakrabarti, A;Schatten, H;Taylor, M
通讯作者：
Taylor, M
DOI：
10.1073/pnas.1735416100
发表时间：
2003-11-11
影响因子：
11.1
作者：
Liedtke, W;Friedman, JM
通讯作者：
Friedman, JM
影响因子：
7.8
作者：
Koetttgen, Michael;Buchholz, Bjoern;Garcia-Gonzalez, Miguel A.;Kotsis, Fruzsina;Fu, Xiao;Doerken, Mara;Boehlke, Christopher;Steffl, Daniel;Tauber, Robert;Wegierski, Tomasz;Nitschke, Roland;Suzuki, Makoto;Kramer-Zucker, Albrecht;Germino, Gregory G.;Watnick, Terry;Prenen, Jean;Nilius, Bernd;Kuehn, E. Wolfgang;Walz, Gerd
通讯作者：
Walz, Gerd