Channelrhodopsin-2 localised to the axon initial segment.
The light-gated cation channel Channelrhodopsin-2 (ChR2) is a powerful and versatile tool for controlling neuronal activity. Currently available versions of ChR2 either distribute uniformly throughout the plasma membrane or are localised specifically to somatodendritic or synaptic domains. Localisin...
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Public Library of Science (PLoS)
2010-10-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0013761&type=printable |
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| author | Matthew S Grubb Juan Burrone |
| author_facet | Matthew S Grubb Juan Burrone |
| author_sort | Matthew S Grubb |
| collection | DOAJ |
| description | The light-gated cation channel Channelrhodopsin-2 (ChR2) is a powerful and versatile tool for controlling neuronal activity. Currently available versions of ChR2 either distribute uniformly throughout the plasma membrane or are localised specifically to somatodendritic or synaptic domains. Localising ChR2 instead to the axon initial segment (AIS) could prove an extremely useful addition to the optogenetic repertoire, targeting the channel directly to the site of action potential initiation, and limiting depolarisation and associated calcium entry elsewhere in the neuron. Here, we describe a ChR2 construct that we localised specifically to the AIS by adding the ankyrinG-binding loop of voltage-gated sodium channels (Na(v)II-III) to its intracellular terminus. Expression of ChR2-YFP-Na(v)II-III did not significantly affect the passive or active electrical properties of cultured rat hippocampal neurons. However, the tiny ChR2 currents and small membrane depolarisations resulting from AIS targeting meant that optogenetic control of action potential firing with ChR2-YFP-Na(v)II-III was unsuccessful in baseline conditions. We did succeed in stimulating action potentials with light in some ChR2-YFP-Na(v)II-III-expressing neurons, but only when blocking KCNQ voltage-gated potassium channels. We discuss possible alternative approaches to obtaining precise control of neuronal spiking with AIS-targeted optogenetic constructs and propose potential uses for our ChR2-YFP-Na(v)II-III probe where subthreshold modulation of action potential initiation is desirable. |
| format | Article |
| id | doaj-art-b5804bbd1ed84ab3920c4f0408bf2db1 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2010-10-01 |
| publisher | Public Library of Science (PLoS) |
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| spelling | doaj-art-b5804bbd1ed84ab3920c4f0408bf2db12025-08-20T02:01:57ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-10-01510e1376110.1371/journal.pone.0013761Channelrhodopsin-2 localised to the axon initial segment.Matthew S GrubbJuan BurroneThe light-gated cation channel Channelrhodopsin-2 (ChR2) is a powerful and versatile tool for controlling neuronal activity. Currently available versions of ChR2 either distribute uniformly throughout the plasma membrane or are localised specifically to somatodendritic or synaptic domains. Localising ChR2 instead to the axon initial segment (AIS) could prove an extremely useful addition to the optogenetic repertoire, targeting the channel directly to the site of action potential initiation, and limiting depolarisation and associated calcium entry elsewhere in the neuron. Here, we describe a ChR2 construct that we localised specifically to the AIS by adding the ankyrinG-binding loop of voltage-gated sodium channels (Na(v)II-III) to its intracellular terminus. Expression of ChR2-YFP-Na(v)II-III did not significantly affect the passive or active electrical properties of cultured rat hippocampal neurons. However, the tiny ChR2 currents and small membrane depolarisations resulting from AIS targeting meant that optogenetic control of action potential firing with ChR2-YFP-Na(v)II-III was unsuccessful in baseline conditions. We did succeed in stimulating action potentials with light in some ChR2-YFP-Na(v)II-III-expressing neurons, but only when blocking KCNQ voltage-gated potassium channels. We discuss possible alternative approaches to obtaining precise control of neuronal spiking with AIS-targeted optogenetic constructs and propose potential uses for our ChR2-YFP-Na(v)II-III probe where subthreshold modulation of action potential initiation is desirable.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0013761&type=printable |
| spellingShingle | Matthew S Grubb Juan Burrone Channelrhodopsin-2 localised to the axon initial segment. PLoS ONE |
| title | Channelrhodopsin-2 localised to the axon initial segment. |
| title_full | Channelrhodopsin-2 localised to the axon initial segment. |
| title_fullStr | Channelrhodopsin-2 localised to the axon initial segment. |
| title_full_unstemmed | Channelrhodopsin-2 localised to the axon initial segment. |
| title_short | Channelrhodopsin-2 localised to the axon initial segment. |
| title_sort | channelrhodopsin 2 localised to the axon initial segment |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0013761&type=printable |
| work_keys_str_mv | AT matthewsgrubb channelrhodopsin2localisedtotheaxoninitialsegment AT juanburrone channelrhodopsin2localisedtotheaxoninitialsegment |