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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
ZUSAMMENFASSUNG

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).

MATERIALIEN
Produktnummer
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Produktbeschreibung

Sigma-Aldrich
HEPES, ≥99.5% (titration)
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D-(+)-Glukose, ≥99.5% (GC)
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Natriumbicarbonat, powder, BioReagent, Molecular Biology, suitable for cell culture, suitable for insect cell culture
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Guanosin 5′-triphosphat Natriumsalz Hydrat, ≥95% (HPLC), powder
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Adenosin-5′-triphosphat Magnesiumsalz, ≥95%, bacterial
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Kaliumchlorid, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
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Ethylenglykol-bis(2-aminoethylether)-N,N,N′,N′-Tetraessigsäure, Molecular Biology, ≥97.0%
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(+)-Natrium-L-ascorbat, crystalline, ≥98%
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Kreatinphosphat Dinatriumsalz Hydrat, ≥97%
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Biocytin, ≥98% (TLC)
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Kaliumhydroxid, BioXtra, ≥85% KOH basis
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Urethan, ≥99%
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QX-314, A quaternary lidocaine derivative, permanently charged, and lipophobic.
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Natriumchlorid, BioXtra, ≥99.5% (AT)
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Natriumpyruvat, ReagentPlus®, ≥99%
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Magnesiumchlorid Hexahydrat, BioXtra, ≥99.0%
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N-Methyl-D-Glucamin, 99.0-100.5% (titration)