Characterization of Na+-K+-2Cl- cotransporter activity in rabbit lacrimal gland duct cells

Eszter Vizvári, Máté Katona, Péter Orvos, Orsolya Berczeli, Andrea Facskó, Ferenc Rárosi, V. Venglovecz, Z. Rakonczay, P. Hegyi, Chuanqing Ding, Edit Tóth-Molnár

Research output: Contribution to journalArticle

Abstract

PURPOSE. We recently reported that isolated duct segments from rabbit lacrimal gland (LG) were able to secrete fluid in response to secretagogues, which were blocked completely by bumetanide. This suggests the functional involvement of Na+-K+-2Cl- cotransporter (NKCC1) in ductal fluid secretion. Therefore, the aim of this study was to investigate the activity profile of NKCC1 in isolated rabbit LG duct segments. METHODS. Interlobular ducts were isolated from fresh rabbit LG tissue. Microfluorometry with the ammonium (NH4 +)–pulse technique was used to elicit pH changes in duct cells, and the rate of bumetanide-sensitive cytosolic acidification after addition of NH4 + was used to quantify the activity of NKCC1. RESULTS. While basal activity of NKCC1 was undetectable, low cytosolic chloride (Cl-) level and hyperosmotic challenge (390 mOsm) were able to increase the activity of NKCC1. Carbachol (100 μM) had no significant effect on NKCC1 activity. Elevation of cytosolic calcium (Ca2+) level with Ca2+-ionophore (A 23187, 1 μM) did not cause any alteration in the activity of the cotransporter while direct activation of protein kinase C (phorbol myristate acetate, 100 nM) increased its activity slightly but in a significant manner. Addition of either forskolin (10 μM), cell-permeable cAMP analogue (8-bromo cAMP, 100 μM) or vasoactive intestinal peptide (200 nM) resulted in a significant increase in the activity of NKCC1. CONCLUSIONS. These results highlight the functional involvement of NKCC1 in LG duct secretion. These findings may facilitate our understanding of LG function and may contribute to the development of targeted pharmacologic interventions in case of dry eye disease.

Original languageEnglish
Pages (from-to)3828-3835
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume57
Issue number8
DOIs
Publication statusPublished - Jul 1 2016

Fingerprint

Lacrimal Apparatus
Rabbits
Bumetanide
Member 2 Solute Carrier Family 12
Cytophotometry
8-Bromo Cyclic Adenosine Monophosphate
Fluids and Secretions
Eye Diseases
Vasoactive Intestinal Peptide
Ionophores
Calcimycin
Carbachol
Tetradecanoylphorbol Acetate
Colforsin
Ammonium Compounds
Protein Kinase C
Chlorides
Calcium

Keywords

  • Duct epithelium
  • Duct epithelium
  • Lacrimal gland
  • Lacrimal gland
  • NKCC1
  • NKCC1

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Vizvári, E., Katona, M., Orvos, P., Berczeli, O., Facskó, A., Rárosi, F., ... Tóth-Molnár, E. (2016). Characterization of Na+-K+-2Cl- cotransporter activity in rabbit lacrimal gland duct cells. Investigative Ophthalmology and Visual Science, 57(8), 3828-3835. https://doi.org/10.1167/iovs.15-18462

Characterization of Na+-K+-2Cl- cotransporter activity in rabbit lacrimal gland duct cells. / Vizvári, Eszter; Katona, Máté; Orvos, Péter; Berczeli, Orsolya; Facskó, Andrea; Rárosi, Ferenc; Venglovecz, V.; Rakonczay, Z.; Hegyi, P.; Ding, Chuanqing; Tóth-Molnár, Edit.

In: Investigative Ophthalmology and Visual Science, Vol. 57, No. 8, 01.07.2016, p. 3828-3835.

Research output: Contribution to journalArticle

Vizvári, Eszter ; Katona, Máté ; Orvos, Péter ; Berczeli, Orsolya ; Facskó, Andrea ; Rárosi, Ferenc ; Venglovecz, V. ; Rakonczay, Z. ; Hegyi, P. ; Ding, Chuanqing ; Tóth-Molnár, Edit. / Characterization of Na+-K+-2Cl- cotransporter activity in rabbit lacrimal gland duct cells. In: Investigative Ophthalmology and Visual Science. 2016 ; Vol. 57, No. 8. pp. 3828-3835.
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AU - Berczeli, Orsolya

AU - Facskó, Andrea

AU - Rárosi, Ferenc

AU - Venglovecz, V.

AU - Rakonczay, Z.

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