Expression of anion exchangers in cultured human endolymphatic sac epithelia

Hyun Jae Lee, Won Sun Yang, Hyun Woo Park, Hyun Seung Choi, Sung Huhn Kim, Jin Young Kim, Jae Young Choi

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

HYPOTHESIS: Pendrin acts as a Cl-/HCO3- exchanger and is responsible for endolymphatic fluid volume and pH homeostasis in human endolymphatic sac epithelial cells. BACKGROUND: The endolymphatic sac (ES) is part of the membranous labyrinth in the inner ear that plays an important role in maintaining homeostasis of the endolymphatic fluid system. However, the exact mechanism of fluid volume and pH regulation is not fully understood yet. We aimed to demonstrate the expression of various anion exchangers (AEs), including pendrin, in cultured human endolymphatic sac epithelial (HESE) cells. METHODS: Endolymphatic sac specimens were harvested during acoustic neuroma surgery (n = 24) using the translabyrinthine approach and then subcultured with high epidermal growth factor (EGF) (25 ng/ml) media and differentiated using low-EGF (0.5 ng/ml) media. The cultured cells were classified according to the morphology on TEM. The Cl-/HCO3- exchanger activity was assessed by pHi measurement using pH sensitive dye 2′, 7′-bis (2-carboxyethyl)-5(6)- carboxyfluorescein (BCECF/AM). We performed reverse transcriptase-polymerase chain reaction and immunohistochemical staining for AEs. RESULTS: We determined that 7.3 ± 6.7% of cells differentiated into mitochodria-rich cells and 50.2 ± 15.1 of cells differentiated into ribosome-rich cells. bAE3, AE4, SLC26A4, SLC26A6, and SLC26A11 were also expressed in cultured HESE cells. The cultured cells had Cl-/HCO3- and Cl-/formate exchange activity on the luminal membrane, which is sensitive to anion channel inhibitors (DIDS 500 μM). Furthermore, we showed that pendrin (SLC26A4) was expressed in cultured HESE cell membranes. CONCLUSION: Our results suggest that AEs, including pendrin, are expressed in epithelia of ES and may have role in maintaining ionic homeostasis, and the HESE culture system are useful for uncovering the functional role of ES epithelial cells.

Original languageEnglish
Pages (from-to)1664-1671
Number of pages8
JournalOtology and Neurotology
Volume33
Issue number9
DOIs
Publication statusPublished - 2012 Dec 1

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Endolymphatic Sac
Anions
Epithelium
Epithelial Cells
Chloride-Bicarbonate Antiporters
formic acid
Homeostasis
Inner Ear
Epidermal Growth Factor
Cultured Cells
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Acoustic Neuroma
Reverse Transcriptase Polymerase Chain Reaction
Ribosomes
Cell Membrane
Staining and Labeling

All Science Journal Classification (ASJC) codes

  • Otorhinolaryngology
  • Clinical Neurology
  • Sensory Systems
  • Medicine(all)

Cite this

Lee, H. J., Yang, W. S., Park, H. W., Choi, H. S., Kim, S. H., Kim, J. Y., & Choi, J. Y. (2012). Expression of anion exchangers in cultured human endolymphatic sac epithelia. Otology and Neurotology, 33(9), 1664-1671. https://doi.org/10.1097/MAO.0b013e31826bf3d3
Lee, Hyun Jae ; Yang, Won Sun ; Park, Hyun Woo ; Choi, Hyun Seung ; Kim, Sung Huhn ; Kim, Jin Young ; Choi, Jae Young. / Expression of anion exchangers in cultured human endolymphatic sac epithelia. In: Otology and Neurotology. 2012 ; Vol. 33, No. 9. pp. 1664-1671.
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abstract = "HYPOTHESIS: Pendrin acts as a Cl-/HCO3- exchanger and is responsible for endolymphatic fluid volume and pH homeostasis in human endolymphatic sac epithelial cells. BACKGROUND: The endolymphatic sac (ES) is part of the membranous labyrinth in the inner ear that plays an important role in maintaining homeostasis of the endolymphatic fluid system. However, the exact mechanism of fluid volume and pH regulation is not fully understood yet. We aimed to demonstrate the expression of various anion exchangers (AEs), including pendrin, in cultured human endolymphatic sac epithelial (HESE) cells. METHODS: Endolymphatic sac specimens were harvested during acoustic neuroma surgery (n = 24) using the translabyrinthine approach and then subcultured with high epidermal growth factor (EGF) (25 ng/ml) media and differentiated using low-EGF (0.5 ng/ml) media. The cultured cells were classified according to the morphology on TEM. The Cl-/HCO3- exchanger activity was assessed by pHi measurement using pH sensitive dye 2′, 7′-bis (2-carboxyethyl)-5(6)- carboxyfluorescein (BCECF/AM). We performed reverse transcriptase-polymerase chain reaction and immunohistochemical staining for AEs. RESULTS: We determined that 7.3 ± 6.7{\%} of cells differentiated into mitochodria-rich cells and 50.2 ± 15.1 of cells differentiated into ribosome-rich cells. bAE3, AE4, SLC26A4, SLC26A6, and SLC26A11 were also expressed in cultured HESE cells. The cultured cells had Cl-/HCO3- and Cl-/formate exchange activity on the luminal membrane, which is sensitive to anion channel inhibitors (DIDS 500 μM). Furthermore, we showed that pendrin (SLC26A4) was expressed in cultured HESE cell membranes. CONCLUSION: Our results suggest that AEs, including pendrin, are expressed in epithelia of ES and may have role in maintaining ionic homeostasis, and the HESE culture system are useful for uncovering the functional role of ES epithelial cells.",
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Expression of anion exchangers in cultured human endolymphatic sac epithelia. / Lee, Hyun Jae; Yang, Won Sun; Park, Hyun Woo; Choi, Hyun Seung; Kim, Sung Huhn; Kim, Jin Young; Choi, Jae Young.

In: Otology and Neurotology, Vol. 33, No. 9, 01.12.2012, p. 1664-1671.

Research output: Contribution to journalArticle

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T1 - Expression of anion exchangers in cultured human endolymphatic sac epithelia

AU - Lee, Hyun Jae

AU - Yang, Won Sun

AU - Park, Hyun Woo

AU - Choi, Hyun Seung

AU - Kim, Sung Huhn

AU - Kim, Jin Young

AU - Choi, Jae Young

PY - 2012/12/1

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N2 - HYPOTHESIS: Pendrin acts as a Cl-/HCO3- exchanger and is responsible for endolymphatic fluid volume and pH homeostasis in human endolymphatic sac epithelial cells. BACKGROUND: The endolymphatic sac (ES) is part of the membranous labyrinth in the inner ear that plays an important role in maintaining homeostasis of the endolymphatic fluid system. However, the exact mechanism of fluid volume and pH regulation is not fully understood yet. We aimed to demonstrate the expression of various anion exchangers (AEs), including pendrin, in cultured human endolymphatic sac epithelial (HESE) cells. METHODS: Endolymphatic sac specimens were harvested during acoustic neuroma surgery (n = 24) using the translabyrinthine approach and then subcultured with high epidermal growth factor (EGF) (25 ng/ml) media and differentiated using low-EGF (0.5 ng/ml) media. The cultured cells were classified according to the morphology on TEM. The Cl-/HCO3- exchanger activity was assessed by pHi measurement using pH sensitive dye 2′, 7′-bis (2-carboxyethyl)-5(6)- carboxyfluorescein (BCECF/AM). We performed reverse transcriptase-polymerase chain reaction and immunohistochemical staining for AEs. RESULTS: We determined that 7.3 ± 6.7% of cells differentiated into mitochodria-rich cells and 50.2 ± 15.1 of cells differentiated into ribosome-rich cells. bAE3, AE4, SLC26A4, SLC26A6, and SLC26A11 were also expressed in cultured HESE cells. The cultured cells had Cl-/HCO3- and Cl-/formate exchange activity on the luminal membrane, which is sensitive to anion channel inhibitors (DIDS 500 μM). Furthermore, we showed that pendrin (SLC26A4) was expressed in cultured HESE cell membranes. CONCLUSION: Our results suggest that AEs, including pendrin, are expressed in epithelia of ES and may have role in maintaining ionic homeostasis, and the HESE culture system are useful for uncovering the functional role of ES epithelial cells.

AB - HYPOTHESIS: Pendrin acts as a Cl-/HCO3- exchanger and is responsible for endolymphatic fluid volume and pH homeostasis in human endolymphatic sac epithelial cells. BACKGROUND: The endolymphatic sac (ES) is part of the membranous labyrinth in the inner ear that plays an important role in maintaining homeostasis of the endolymphatic fluid system. However, the exact mechanism of fluid volume and pH regulation is not fully understood yet. We aimed to demonstrate the expression of various anion exchangers (AEs), including pendrin, in cultured human endolymphatic sac epithelial (HESE) cells. METHODS: Endolymphatic sac specimens were harvested during acoustic neuroma surgery (n = 24) using the translabyrinthine approach and then subcultured with high epidermal growth factor (EGF) (25 ng/ml) media and differentiated using low-EGF (0.5 ng/ml) media. The cultured cells were classified according to the morphology on TEM. The Cl-/HCO3- exchanger activity was assessed by pHi measurement using pH sensitive dye 2′, 7′-bis (2-carboxyethyl)-5(6)- carboxyfluorescein (BCECF/AM). We performed reverse transcriptase-polymerase chain reaction and immunohistochemical staining for AEs. RESULTS: We determined that 7.3 ± 6.7% of cells differentiated into mitochodria-rich cells and 50.2 ± 15.1 of cells differentiated into ribosome-rich cells. bAE3, AE4, SLC26A4, SLC26A6, and SLC26A11 were also expressed in cultured HESE cells. The cultured cells had Cl-/HCO3- and Cl-/formate exchange activity on the luminal membrane, which is sensitive to anion channel inhibitors (DIDS 500 μM). Furthermore, we showed that pendrin (SLC26A4) was expressed in cultured HESE cell membranes. CONCLUSION: Our results suggest that AEs, including pendrin, are expressed in epithelia of ES and may have role in maintaining ionic homeostasis, and the HESE culture system are useful for uncovering the functional role of ES epithelial cells.

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