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Regulatory Roles of Anoctamin-6 in Human Trabecular Meshwork Cells.

Publication ,  Journal Article
Banerjee, J; Leung, C-T; Li, A; Peterson-Yantorno, K; Ouyang, H; Stamer, WD; Civan, MM
Published in: Invest Ophthalmol Vis Sci
January 1, 2017

PURPOSE: Trabecular meshwork (TM) cell volume is a determinant of aqueous humor outflow resistance, and thereby IOP. Regulation of TM cell volume depends on chloride ion (Cl-) release through swelling-activated channels (ICl,Swell), whose pore is formed by LRRC8 proteins. Chloride ion release through swelling-activated channels has been reported to be regulated by calcium-activated anoctamins, but this finding is controversial. Particularly uncertain has been the effect of anoctamin Ano6, reported as a Ca2+-activated Cl- (CaCC) or cation channel in other cells. The current study tested whether anoctamin activity modifies volume regulation of primary TM cell cultures and cell lines. METHODS: Gene expression was studied with quantitative PCR, supplemented by reverse-transcriptase PCR and Western immunoblots. Currents were measured by ruptured whole-cell patch clamping and volume by electronic cell sizing. RESULTS: Primary TM cell cultures and the TM5 and GTM3 cell lines expressed Ano6 3 to 4 orders of magnitude higher than the other anoctamin CaCCs (Ano1 and Ano2). Ionomycin increased cell Ca2+ and activated macroscopic currents conforming to CaCCs in other cells, but displayed significantly more positive mean reversal potentials (+5 to +12 mV) than those displayed by ICl,Swell (-14 to -21 mV) in the same cells. Nonselective CaCC inhibitors (tannic acid>CaCCinh-A01) and transient Ano6 knockdown strongly inhibited ionomycin-activated currents, ICl,Swell and the regulatory volume response to hyposmotic swelling. CONCLUSIONS: Ionomycin activates CaCCs associated with net cation movement in TM cells. These currents, ICl,Swell, and cell volume are regulated by Ano6. The findings suggest a novel clinically-relevant approach for altering cell volume, and thereby outflow resistance, by targeting Ano6.

Duke Scholars

Published In

Invest Ophthalmol Vis Sci

DOI

EISSN

1552-5783

Publication Date

January 1, 2017

Volume

58

Issue

1

Start / End Page

492 / 501

Location

United States

Related Subject Headings

  • Trabecular Meshwork
  • Reverse Transcriptase Polymerase Chain Reaction
  • Real-Time Polymerase Chain Reaction
  • Phospholipid Transfer Proteins
  • Patch-Clamp Techniques
  • Ophthalmology & Optometry
  • Humans
  • Gene Expression Regulation
  • DNA
  • Chloride Channels
 

Citation

APA
Chicago
ICMJE
MLA
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Banerjee, J., Leung, C.-T., Li, A., Peterson-Yantorno, K., Ouyang, H., Stamer, W. D., & Civan, M. M. (2017). Regulatory Roles of Anoctamin-6 in Human Trabecular Meshwork Cells. Invest Ophthalmol Vis Sci, 58(1), 492–501. https://doi.org/10.1167/iovs.16-20188
Banerjee, Juni, Chi-Ting Leung, Ang Li, Kim Peterson-Yantorno, Huan Ouyang, W Daniel Stamer, and Mortimer M. Civan. “Regulatory Roles of Anoctamin-6 in Human Trabecular Meshwork Cells.Invest Ophthalmol Vis Sci 58, no. 1 (January 1, 2017): 492–501. https://doi.org/10.1167/iovs.16-20188.
Banerjee J, Leung C-T, Li A, Peterson-Yantorno K, Ouyang H, Stamer WD, et al. Regulatory Roles of Anoctamin-6 in Human Trabecular Meshwork Cells. Invest Ophthalmol Vis Sci. 2017 Jan 1;58(1):492–501.
Banerjee, Juni, et al. “Regulatory Roles of Anoctamin-6 in Human Trabecular Meshwork Cells.Invest Ophthalmol Vis Sci, vol. 58, no. 1, Jan. 2017, pp. 492–501. Pubmed, doi:10.1167/iovs.16-20188.
Banerjee J, Leung C-T, Li A, Peterson-Yantorno K, Ouyang H, Stamer WD, Civan MM. Regulatory Roles of Anoctamin-6 in Human Trabecular Meshwork Cells. Invest Ophthalmol Vis Sci. 2017 Jan 1;58(1):492–501.

Published In

Invest Ophthalmol Vis Sci

DOI

EISSN

1552-5783

Publication Date

January 1, 2017

Volume

58

Issue

1

Start / End Page

492 / 501

Location

United States

Related Subject Headings

  • Trabecular Meshwork
  • Reverse Transcriptase Polymerase Chain Reaction
  • Real-Time Polymerase Chain Reaction
  • Phospholipid Transfer Proteins
  • Patch-Clamp Techniques
  • Ophthalmology & Optometry
  • Humans
  • Gene Expression Regulation
  • DNA
  • Chloride Channels