Abstract
Optogenetics employs engineered viruses to genetically modify cells to express specific
light-sensitive ion channels. The standard method for gene delivery in the brain involves
invasive craniotomies that expose the brain and direct injections of viruses that
invariably damage neural tissue along the syringe tract. A recently proposed alternative
in which non-invasive optogenetics is performed with focused ultrasound (FUS)–mediated
blood–brain barrier (BBB) openings has been found to non-invasively facilitate gene
delivery for optogenetics in mice. Although gene delivery can be performed non-invasively,
validating successful viral transduction and expression of encoded ion channels in
target tissue typically involves similar invasive techniques, such as craniotomies
in longitudinal studies and/or postmortem histology. Functional ultrasound imaging
(fUSi) is an emerging neuroimaging technique that can be used to transcranially detect
changes in cerebral blood volume following introduction of a stimulus. In this study,
we implemented a fully non-invasive combined FUS–fUSi technique for performing optogenetics
in mice. FUS successfully delivered viruses encoding the red-shifted channelrhodopsin
variant ChrimsonR in all treated subjects. fUSi successfully identified stimulus-evoked
cerebral blood volume changes preferentially in brain regions expressing the light-sensitive
ion channels. Improvements in cell-specific targeting of viral vectors and transcranial
ultrasound imaging will make the combined technique a useful tool for neuroscience
research in small animals.
Key Words
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References
- An optical neural interface: In vivo control of rodent motor cortex with integrated fiberoptic and optogenetic technology.J Neural Eng. 2007; 4: 143-156
- Millisecond-timescale, genetically targeted optical control of neural activity.Nat Neurosci. 2005; 8: 1263-1268
- Whole-brain functional ultrasound imaging in awake head-fixed mice.Nat Protoc. 2021; 16: 3547-3571
- Near-infrared deep brain stimulation via upconversion nanoparticle–mediated optogenetics.Science. 2018; 359: 679-684
- Noninvasive, transcranial and localized opening of the blood–brain barrier using focused ultrasound in mice.Ultrasound Med Biol. 2007; 33: 95-104
- Noninvasive optical inhibition with a red-shifted microbial rhodopsin.Nat Neurosci. 2014; 17: 1123-1129
- Simultaneous electroencephalography, real-time measurement of lactate concentration and optogenetic manipulation of neuronal activity in the rodent cerebral cortex.J Vis Exp. 2012; 70
- Spatiotemporal clutter filtering of ultrafast ultrasound data highly increases Doppler and fUltrasound sensitivity.IEEE Trans Med Imaging. 2015; 34: 2271-2285
- Chemogenetics revealed: DREADD occupancy and activation via converted clozapine.Science. 2017; 357: 503-507
- Sonogenetics is a non-invasive approach to activating neurons in Caenorhabditis elegans.Nat Commun. 2015; 6: 1-12
- Focused ultrasound neuromodulation of cortical and subcortical brain structures using 1.9 MHz.Medical Physics. 2016; 43: 5730-5735https://doi.org/10.1118/1.4963208
- Limitations of temporal resolution in functional MRI.Magn Reson Med. 1997; 37: 631-636
- Conditional modification of behavior in Drosophila by targeted expression of a temperature-sensitive shibire allele in defined neurons.J Neurobiol. 2001; 47: 81-92
- Independent optical excitation of distinct neural populations.Nat Methods. 2014; 11: 338-346
- High-density EEG recordings of the freely moving mice using polyimide-based microelectrode.J Vis Exp. 2011; 47: 2562
- Optogenetic fmri interrogation of brain-wide central vestibular pathways.Proc Natl Acad Sci USA. 2019; 116: 10122-10129
- ReaChR: A red-shifted variant of Channelrhodopsin enables deep transcranial optogenetic excitation.Nat Neurosci. 2013; 16: 1499-1508
- Functional ultrasound imaging of the brain.Nat Methods. 2011; 8: 662-664
- Biomolecular ultrasound and sonogenetics.Annu Rev Chem Biomol Eng. 2018; 9: 229-252
- Non-invasive optogenetics with ultrasond-mediated gene delivery and red-light excitation.Brain Stimulation. 2022; 15: 927-941
- Optogenetic fUSI for brain-wide mapping of neural activity mediating collicular-dependent behaviors.Neuron. 2021; 109 (e10): 1888-1905
- Tapered fibers combined with a multi-electrode array for optogenetics in mouse medial prefrontal cortex.Front Neurosci. 2018; 12: 771
- Simultaneous optogenetics and cellular resolution calcium imaging during active behavior using a miniaturized microscope.Front Neurosci. 2018; 12: 496
- Real-time imaging of brain activity in freely moving rats using functional ultrasound.Nat Methods. 2015; 12: 873-878
- Noninvasive, neuron-specific gene therapy can be facilitated by focused ultrasound and recombinant adeno-associated virus.Gene Ther. 2014; 22: 104-110
- Non-invasive, focused ultrasound-facilitated gene delivery for optogenetics.Sci Rep. 2017; 7: 39955
- Sonothermogenetics for noninvasive and cell-type specific deep brain neuromodulation.Brain Stimul. 2021; 14: 790-800
- Red-shifted optogenetic excitation: a tool for fast neural control derived from Volvox carteri.Nat Neurosci. 2008; 11: 631-633
Article info
Publication history
Published online: November 30, 2022
Accepted:
November 3,
2022
Received in revised form:
October 27,
2022
Received:
January 11,
2022
Publication stage
In Press Corrected ProofIdentification
Copyright
© 2022 World Federation for Ultrasound in Medicine & Biology. All rights reserved.
