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Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels
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Calcium-Activated Chloride Channels in Newly Differentiating Mouse Lens Fiber Cells and Their Role in Volume
Jun-Jie Tong1, Pooja Acharya1, Lisa Ebihara1
1Department of Physiology and Biophysics, Rosalind Franklin University of Medicine and Science, Chicago, Illinois, United States.
Investigative Ophthalmology & Visual Science
|April 18, 2019
Summary
Newly differentiating mouse lens fiber cells express calcium-activated chloride channels (CaCCs), specifically TMEM16A and TMEM16B, which are crucial for regulating lens volume.
Area of Science:
- Ocular physiology
- Ion channel research
- Cell biology
Background:
- Chloride channels are vital for lens volume regulation.
- Identifying lens fiber cell channels is challenging due to isolation difficulties.
- A novel technique allows isolation of mouse fiber cells lacking Cx50 and Cx46.
Purpose of the Study:
- To investigate if newly differentiating mouse fiber cells express calcium-activated chloride channels (CaCCs).
- To utilize a new cell isolation technique for this purpose.
Main Methods:
- Isolated differentiating fiber cells from Cx50/Cx46 double knockout mice.
- Employed whole-cell patch clamp for electrophysiology.
- Used RT-PCR and immunofluorescence to identify CaCC molecular identity and distribution.
Main Results:
- Peripheral fiber cells exhibit calcium-activated chloride currents.
- These currents share properties with TMEM16 CaCCs.
- TMEM16A and TMEM16B transcripts and proteins were found in differentiating lens cells.
Conclusions:
- Peripheral fiber cells possess CaCCs attributed to TMEM16A and TMEM16B.
- These channels likely play a significant role in lens volume regulation, similar to other tissues.
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