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Controllable Ion Channel Expression through Inducible Transient Transfection
Published on: February 17, 2017
Cambios rápidos inducidos químicamente de los canales iónicos de la puerta KCNQ de PtdIns(4,5) P2 de la puerta KCNQ
Byung-Chang Suh1, Takanari Inoue, Tobias Meyer
1Department of Physiology and Biophysics, University of Washington School of Medicine, Seattle, WA 98195, USA.
Science (New York, N.Y.)
|September 23, 2006
Resumen
El agotamiento del fosfatidilinositol 4,5-bisfosfato (PI(4,5) P2) suprime completamente la corriente del canal iónico KCNQ, independiente de otros mensajeros. Este hallazgo aclara las vías de señalización y resalta PI(4,5)P2
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Fisiología celular Fisiología celular.
- La bioquímica es la bioquímica.
Sus antecedentes:
- Los canales iónicos KCNQ son cruciales para la excitabilidad neuronal.
- La activación de la fosfolipasa C (PLC) conduce a la supresión de la corriente KCNQ, pero los mensajeros específicos involucrados siguen siendo controvertidos.
- Los fosfoinosítidos, en particular el fosfatidilinositol 4,5-bisfosfato (PI(4,5) P2), son reguladores clave de las proteínas de la membrana.
Objetivo del estudio:
- Para identificar definitivamente los segundos mensajeros responsables de la supresión de corriente KCNQ mediada por PLC.
- Investigar el papel directo de PI{4,5) P2 en la regulación de la actividad del canal KCNQ.
Principales métodos:
- Desarrollo de enzimas translocables para manipular rápidamente los niveles de fosfoinosítidos de la membrana plasmática tras la aplicación de señales químicas.
- El agotamiento sistemático y la sobreproducción de fosfoinosítidos específicos, incluidos el PI{4,5) P2 y el fosfatidilinositol 3,4,5-trisfosfato (PI{3,4,5) P3).
- Medición en tiempo real de la corriente del canal iónico KCNQ en respuesta a los cambios en el nivel de fosfoinosítidos, mientras se monitorean otros mensajeros potenciales como Ca2+, diacilglicerol e inositol 1,4,5-trisfosfato.
Principales resultados:
- La corriente de KCNQ se suprimió rápidamente al agotamiento de PI{4,5) P2, independientemente de los cambios en los niveles de Ca2+, diacilglicerol o inositol 1,4,5-trisfosfato.
- La sobreproducción de PI{4,5) P2 dio lugar a un aumento del 30% en el KCNQ actual.
- La modulación de los niveles de PI{3,4,5) P3 no afectó a la corriente KCNQ.
Conclusiones:
- El agotamiento de PI{4,5) P2 es suficiente para suprimir completamente la corriente del canal iónico KCNQ.
- Otros segundos mensajeros comúnmente asociados con la señalización PLC no son necesarios para esta supresión.
- El enfoque enzimático desarrollado proporciona una poderosa herramienta para estudiar las redes de señalización mediadas por fosfoinositida.
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