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TMEM16A alternative splicing isoforms in Xenopus tropicalis: distribution and functional properties
A Huanosta-Gutiérrez1, A E Espino-Saldaña1, J P Reyes1
1Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Instituto de Neurobiología, Campus UNAM Juriquilla, Querétaro, Qro CP 76230, Mexico.
Biochemical and Biophysical Research Communications
|March 26, 2014
Summary
Xenopus tropicalis oocytes express TMEM16A, a calcium-dependent chloride channel. This channel generates a unique current distinct from previously identified voltage-dependent currents in these cells.
Area of Science:
- Molecular Biology
- Ion Channel Physiology
- Xenopus Oocyte Research
Background:
- Xenopus tropicalis oocytes exhibit a calcium-dependent chloride current (ICl,Ca).
- This current shares characteristics with TMEM16A ion channel activity.
- Previous studies identified voltage-dependent chloride currents in these oocytes.
Purpose of the Study:
- To investigate the role of TMEM16A in Xenopus tropicalis oocyte chloride currents.
- To characterize the functional properties of TMEM16A isoforms expressed in oocytes.
- To differentiate TMEM16A-mediated currents from other known oocyte currents.
Main Methods:
- Reverse transcription polymerase chain reaction (RT-PCR) to identify TMEM16A isoforms.
- Heterologous expression of TMEM16A isoforms in HEK-293 cells.
- Electrophysiological recordings (patch-clamp) to analyze chloride channel activity.
Main Results:
- Five alternatively spliced TMEM16A isoforms were detected in Xenopus tropicalis oocytes.
- Expressed TMEM16A isoforms formed functional calcium-dependent chloride channels.
- A specific exon (exon 1d) correlated with a transient current upon hyperpolarization.
Conclusions:
- Xenopus tropicalis oocytes express TMEM16A, contributing to calcium-dependent chloride currents.
- These TMEM16A-mediated currents are distinct from previously described voltage-dependent currents.
- Differential expression of TMEM16A isoforms suggests tissue-specific roles in Xenopus.
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