Pflügers Archiv - European Journal of Physiology

, Volume 468, Issue 10, pp 1751–1763 | Cite as

Relationship between TMEM16A/anoctamin 1 and LRRC8A

  • Roberta Benedetto
  • Lalida Sirianant
  • Ines Pankonien
  • Podchanart Wanitchakool
  • Jiraporn Ousingsawat
  • Ines Cabrita
  • Rainer Schreiber
  • Margarida Amaral
  • Karl KunzelmannEmail author
Signaling and cell physiology


TMEM16A/anoctamin 1/ANO1 and VRAC/LRRC8 are independent chloride channels activated either by increase in intracellular Ca2+ or cell swelling, respectively. In previous studies, we observed overlapping properties for both types of channels. (i) TMEM16A/ANO1 and LRRC8 are inhibited by identical compounds, (ii) the volume-regulated anion channel VRAC requires compartmentalized Ca2+ increase to be fully activated, (iii) anoctamins are activated by cell swelling, (iv) both channels have a role for apoptotic cell death, (v) both channels are possibly located in lipid rafts/caveolae like structures, and (vi) VRAC and anoctamin 1 currents are not additive when each are fully activated. In the present study, we demonstrate in different cell types that loss of LRRC8A expression not only inhibited VRAC, but also attenuated Ca2+ activated Cl currents. Moreover, expression of LRRC8A enhanced Ca2+ activated Cl currents, and both LRRC8A and ANO1 could be coimmunoprecipitated. We found that LRRC8A becomes accessible to biotinylation upon exposure to hypotonic bath solution, while membrane capacitance was not enhanced. When intracellular Ca2+ was increased in ANO1-expressing cells, the membrane capacitance was enhanced and increased binding of FM4–64 to the membrane was observed. As this was not seen in cells lacking ANO1 expression, a role of ANO1 for exocytosis was suggested. We propose that ANO1 and LRRC8A are activated in parallel. Thus, ionomycin or purinergic stimulation will not only activate ANO1 but also LRRC8 currents. Cell swelling will not only activate LRRC8/VRAC, but also stimulate ANO1 currents by enhancing compartmentalized Ca2+ increase and/or through swelling induced autocrine release of ATP.


LRRC8A VRAC Volume-activated anion channels Ca2+-activated chloride currents TMEM16A Anoctamin 1 



This study is supported by DFG SFB699-A7/A12, DFG KU756/12-1, Medical Faculty of Kiel University (F355910 to SK), DFG SFB699-A7/A12, and DFG KU756/12-1. HCT-wt (LRRC8A+/+) and HCT-LRRC8A-/- cells were kindly provided by Dr. Voss/Prof. Dr. Jentsch (FMP, Berlin). The excellent technical assistance by Ms. B. Wild, P. Seeberger, and E. Tartler is gratefully acknowledged.

Supplementary material

424_2016_1862_MOESM1_ESM.pdf (171 kb)
Supplementary Figure 1 (PDF 170 kb)
424_2016_1862_MOESM2_ESM.pdf (115 kb)
Supplementary Figure 2 (PDF 114 kb)
424_2016_1862_MOESM3_ESM.pdf (345 kb)
Supplementary Figure 3 (PDF 345 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Roberta Benedetto
    • 1
  • Lalida Sirianant
    • 1
  • Ines Pankonien
    • 2
  • Podchanart Wanitchakool
    • 1
  • Jiraporn Ousingsawat
    • 1
  • Ines Cabrita
    • 1
  • Rainer Schreiber
    • 1
  • Margarida Amaral
    • 2
  • Karl Kunzelmann
    • 1
    Email author
  1. 1.Physiological InstituteUniversity of RegensburgRegensburgGermany
  2. 2.Faculty of Sciences, Biosystems & Integrative Sciences InstituteUniversity of LisboaLisbonPortugal

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