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A method for measuring electrical signals in a primary cilium

BACKGROUND: Most cells in the body possess a single primary cilium. These cilia are key transducers of sensory stimuli, and defects in cilia have been linked to several diseases. Evidence suggests that some transduction of sensory stimuli by the primary cilium depends on ion-conducting channels. How...

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Autores principales: Kleene, Nancy K, Kleene, Steven J
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3539729/
https://www.ncbi.nlm.nih.gov/pubmed/23308345
http://dx.doi.org/10.1186/2046-2530-1-17
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author Kleene, Nancy K
Kleene, Steven J
author_facet Kleene, Nancy K
Kleene, Steven J
author_sort Kleene, Nancy K
collection PubMed
description BACKGROUND: Most cells in the body possess a single primary cilium. These cilia are key transducers of sensory stimuli, and defects in cilia have been linked to several diseases. Evidence suggests that some transduction of sensory stimuli by the primary cilium depends on ion-conducting channels. However, the tiny size of the cilium has been a critical barrier to understanding its electrical properties. We report a novel method that allows sensitive, repeatable electrical recordings from primary cilia. Adherent cells were grown on small, spherical beads that could be easily moved within the recording chamber. In this configuration, an entire cilium could be pulled into a recording microelectrode. RESULTS: In 47% of attempts, suction resulted in a seal with high input resistance. Single channels could be recorded while the cilium remained attached to the cell. When the pipette was raised into the air, the cell body was pulled off at the air-bath interface. The pipette retained the cilium and could then be immersed in various solutions that bathed the cytoplasmic face of the membrane. In excised cilia, ionic currents through ciliary channels were modulated by cytoplasmic Ca(2+) and transmembrane voltage. CONCLUSIONS: Ciliary recording is a direct way to learn the effects of second messengers and voltage changes on ciliary transduction channels.
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spelling pubmed-35397292013-01-08 A method for measuring electrical signals in a primary cilium Kleene, Nancy K Kleene, Steven J Cilia Methodology BACKGROUND: Most cells in the body possess a single primary cilium. These cilia are key transducers of sensory stimuli, and defects in cilia have been linked to several diseases. Evidence suggests that some transduction of sensory stimuli by the primary cilium depends on ion-conducting channels. However, the tiny size of the cilium has been a critical barrier to understanding its electrical properties. We report a novel method that allows sensitive, repeatable electrical recordings from primary cilia. Adherent cells were grown on small, spherical beads that could be easily moved within the recording chamber. In this configuration, an entire cilium could be pulled into a recording microelectrode. RESULTS: In 47% of attempts, suction resulted in a seal with high input resistance. Single channels could be recorded while the cilium remained attached to the cell. When the pipette was raised into the air, the cell body was pulled off at the air-bath interface. The pipette retained the cilium and could then be immersed in various solutions that bathed the cytoplasmic face of the membrane. In excised cilia, ionic currents through ciliary channels were modulated by cytoplasmic Ca(2+) and transmembrane voltage. CONCLUSIONS: Ciliary recording is a direct way to learn the effects of second messengers and voltage changes on ciliary transduction channels. BioMed Central 2012-09-03 /pmc/articles/PMC3539729/ /pubmed/23308345 http://dx.doi.org/10.1186/2046-2530-1-17 Text en Copyright ©2012 Kleene and Kleene; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology
Kleene, Nancy K
Kleene, Steven J
A method for measuring electrical signals in a primary cilium
title A method for measuring electrical signals in a primary cilium
title_full A method for measuring electrical signals in a primary cilium
title_fullStr A method for measuring electrical signals in a primary cilium
title_full_unstemmed A method for measuring electrical signals in a primary cilium
title_short A method for measuring electrical signals in a primary cilium
title_sort method for measuring electrical signals in a primary cilium
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3539729/
https://www.ncbi.nlm.nih.gov/pubmed/23308345
http://dx.doi.org/10.1186/2046-2530-1-17
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