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TRP Channels Localize to Subdomains of the Apical Plasma Membrane in Human Fetal Retinal Pigment Epithelium.

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WOS被引频次:11
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成果类型:
期刊论文
作者:
Zhao, Peter Y.;Gan, Geliang;Peng, Shaomin;Wang, Shao-Bin;Chen, Bo;Adelman, Ron A.;Rizzolo, Lawrence J.*
通讯作者:
Rizzolo, Lawrence J.
作者机构:
[Peng, Shaomin; Wang, Shao-Bin; Zhao, Peter Y.; Rizzolo, Lawrence J.; Gan, Geliang] Yale Univ, Dept Surg, New Haven, CT 06520 USA.
[Peng, Shaomin; Wang, Shao-Bin; Rizzolo, Lawrence J.; Chen, Bo; Adelman, Ron A.] Yale Univ, Dept Ophthalmol & Visual Sci, New Haven, CT 06520 USA.
[Peng, Shaomin] Cent S Univ, Aier Sch Ophthalmol, Changsha, Hunan, Peoples R China.
[Rizzolo, Lawrence J.] Yale Univ, Dept Surg, POB 208062, New Haven, CT 06520 USA.
通讯机构:
[Rizzolo, Lawrence J.] Yale Univ, Dept Surg, POB 208062, New Haven, CT 06520 USA.
语种:
英文
关键词:
TRP channels;retinal pigment epithelium;subretinal space;tight junction
期刊:
Investigative ophthalmology & visual science
ISSN:
0146-0404
年:
2015
卷:
56
期:
3
页码:
1916-23
文献类别:
WOS:Article
所属学科:
ESI学科类别:临床医学;WOS学科类别:Ophthalmology
入藏号:
基金类别:
Connecticut Regenerative Medicine Research Fund [10-SCB-02]; Alonzo Family Fund; NIH [CTSA-TL1 TR000141]; International Retinal Research Foundation, Callahan Award; National Natural Science Foundation of China [30772381]; Newman's Own Foundation; Leir Foundation
机构署名:
本校为其他机构
院系归属:
爱尔眼科学院
摘要:
PURPOSE. Calcium regulates many functions of the RPE. Its concentration in the subretinal space and RPE cytoplasm is closely regulated. Transient receptor potential (TRP) channels are a superfamily of ion channels that are moderately calcium-selective. This study investigates the subcellular localization and potential functions of TRP channels in a first-passage culture model of human fetal RPE (hfRPE). METHODS. The RPE isolated from 15- to 16-week gestation fetuses were maintained in serum-free media. Cultures were treated with barium chloride (BaCl2) in the absence and presence of TRP channel inhibitors and monitored by the transepithelial electrical resistance (TER). The expression of TRP channels was determined using quantitative RT-PCR, immunoblotting, and immunofluorescence confocal microscopy. RESULTS. Barium chloride substantially decreased TER and disrupted cell-cell contacts when added to the apical surface of RPE, but not when added to the basolateral surface. The effect could be partially blocked by the general TRP inhibitor, lanthanum chloride (LaCl3, similar to 75%), or an inhibitor of calpain (similar to 25%). Family member-specific inhibitors, ML204 (TRPC4) and HC-067047 (TRPV4), had no effect on basal channel activity. Expression of TRPC4, TRPM1, TRPM3, TRPM7, and TRPV4 was detected by RT-PCR and immunoblotting. The TRPM3 localized to the base of the primary cilium, and TRPC4 and TRPM3 localized to apical tight junctions. The TRPV4 localized to apical microvilli in a small subset of cells. CONCLUSIONS. The TRP channels localized to subdomains of the apical membrane, and BaCl2 was only able to dissociate tight junctions when presented to the apical membrane. The data suggest a potential role for TRP channels as sensors of [Ca2+] in the subretinal space.
参考文献:
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Bozym RA, 2011, MOL BIOL CELL, V22, P3010, DOI 10.1091/mbc.E11-02-0094
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