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  • Bioluminescent optogenetic (BL-OG) activation of neurons during mouse postnatal brain development.

Bioluminescent optogenetic (BL-OG) activation of neurons during mouse postnatal brain development.

STAR protocols (2021-07-22)
Emmanuel L Crespo, Mansi Prakash, Andreas Bjorefeldt, William E Medendorp, Nathan C Shaner, Diane Lipscombe, Christopher I Moore, Ute Hochgeschwender
ABSTRACT

Bioluminescent optogenetics (BL-OG) allows activation of photosensory proteins, such as opsins, by either fiberoptics or by administering a luciferin. BL-OG thus confers both optogenetic and chemogenetic access within the same genetically targeted neuron. This bimodality offers a powerful approach for non-invasive chemogenetic manipulation of neural activity during brain development and adult behaviors with standard optogenetic spatiotemporal precision. We detail protocols for bioluminescent stimulation of neurons in postnatally developing brain and its validation through bioluminescence imaging and electrophysiological recording in mice. For complete information on the use and execution of this protocol, please refer to Medendorp et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium pyruvate, ReagentPlus®, ≥99%
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Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, for molecular biology, ≥97.0%
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Sodium bicarbonate, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
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(+)-Sodium L-ascorbate, crystalline, ≥98%
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Biocytin, ≥98% (TLC)
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Guanosine 5′-triphosphate sodium salt hydrate, ≥95% (HPLC), powder
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Sodium chloride, BioXtra, ≥99.5% (AT)
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HEPES, ≥99.5% (titration)
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D-(+)-Glucose, ≥99.5% (GC)
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Potassium chloride, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
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Adenosine 5′-triphosphate magnesium salt, ≥95%, bacterial
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Magnesium chloride hexahydrate, BioXtra, ≥99.0%
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Urethane, ≥99%
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Phosphocreatine disodium salt hydrate, ≥97%
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N-Methyl-D-glucamine, 99.0-100.5% (titration)
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Potassium hydroxide, BioXtra, ≥85% KOH basis