Optical technologies allowing modulation of neuronal activity at high spatio-temporal resolution are becoming paramount in neuroscience. In this respect, azobenzene-based photoswitches are promising nanoscale tools for neuronal photostimulation. Here we engineered a light-sensitive azobenzene compound (Ziapin2) that stably partitions into the plasma membrane and causes its thinning through trans-dimerization in the dark, resulting in an increased membrane capacitance at steady state. We demonstrated that in neurons loaded with the compound, millisecond pulses of visible light induce a transient hyperpolarization followed by a delayed depolarization that triggers action potential firing. These effects are persistent and can be evoked in vivo up to 7 days, proving the potential of Ziapin2 for the modulation of membrane capacitance in the millisecond timescale, without directly affecting ion channels or local temperature.
Subscribe to Journal
Get full journal access for 1 year
only $15.58 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
The datasets generated and analysed during the current study are available from the corresponding author on reasonable request.
Paoletti, P., Ellis-Davies, G. C. R. & Mourot, A. Optical control of neuronal ion channels and receptors. Nat. Rev. Neurosci. 20, 514–532 (2019).
Rivnay, J., Wang, H., Fenno, L., Deisseroth, K. & Malliaras, G. Next-generation probes, particles, and proteins for neural interfacing. Sci. Adv. 3, e1601649 (2017).
Tønnesen, J. Optogenetic cell control in experimental models of neurological disorders. Behav. Brain Res. 255, 35–43 (2013).
Deisseroth, K. Optogenetics: 10 years of microbial opsins in neuroscience. Nat. Neurosci. 18, 1213–1225 (2015).
Zhang, J. J., Wang, J. X. & Tia, H. Taking orders from light: progress in photochromic bio-materials. Mater. Horiz. 1, 169–184 (2014).
Fujiwara, H. & Yonezawa, Y. Photoelectric response of a black lipid membrane containing an amphiphilic azobenzene derivative. Nature 351, 724–726 (1991).
Yonezawa, Y., Fujiwara, H. & Sato, T. Photoelectric response of black lipid membranes incorporating an amphiphilic azobenzene derivative. Thin Solid Films 210/211, 736–738 (1992).
Tanaka, M. & Yonezawa, Y. Photochemical regulation of ion transport through “quasi-channels” embedded in black lipid membrane. Mat. Sci. Eng. C. 4, 297–301 (1997).
Garner, L. E. et al. Modification of the optoelectronic properties of membranes via insertion of amphiphilic phenylenevinylene oligoelectrolytes. JACS 132, 10042–10052 (2010).
Hinks, J. et al. Modeling cell membrane perturbation by molecules designed for transmembrane electron transfer. Langmuir 30, 2429–2440 (2014).
Gorostiza, P. & Isacoff, E. Optical switches and triggers for the manipulation of ion channels and pores. Mol. Biosyst. 3, 686–704 (2007).
Fortin, D. L. et al. Photochemical control of endogenous ion channels and cellular excitability. Nat. Methods 5, 331–338 (2008).
Kramer, R. H., Mourot, A. & Adesnik, H. Optogenetic pharmacology for control of native neuronal signaling proteins. Nat. Neurosci. 16, 816–823 (2013).
Tochitsky, I., Kienzler, M. A., Isacoff, E. & Kramer, R. H. Restoring vision to the blind with chemical photoswitches. Chem. Rev. 118, 10748–10773 (2018).
Laprell, L. et al. Restoring light sensitivity in blind retinae using a photochromic AMPA receptor agonist. ACS Chem. Neurosci. 7, 15–20 (2016).
Laprell, L. et al. Photopharmacological control of bipolar cells restores visual function in blind mice. J. Clin. Invest. 127, 2598–2611 (2017).
Ghezzi, D. et al. A hybrid bioorganic interface for neuronal photoactivation. Nat. Commun. 2, 166 (2011).
Ghezzi, D. et al. A polymer optoelectronic interface restores light sensitivity in blind rat retinas. Nat. Photonics 7, 400–406 (2013).
Feyen, P. et al. Light-evoked hyperpolarization and silencing of neurons by conjugated polymers. Sci. Rep. 6, 22718 (2016).
Rand, D. et al. Direct electrical neurostimulation with organic pigment photocapacitors. Adv. Mater. 30, e1707292 (2018).
Maya-Vetencourt, J. F. et al. A fully organic retinal prosthesis restores vision in a rat model of degenerative blindness. Nat. Mater. 16, 681–689 (2017).
Martino, N. et al. Photothermal cellular stimulation in functional bio-polymer interfaces. Sci. Rep. 5, 8911 (2015).
Lodola, F., Martino, N., Tullii, G., Lanzani, G. & Antognazza, M. R. Conjugated polymers mediate effective activation of the mammalian ion channel transient receptor potential vanilloid 1. Sci. Rep. 7, 8477 (2017).
Shapiro, M. G., Homma, K., Villarreal, S., Richter, C. P. & Bezanilla, F. Infrared light excites cells by changing their electrical capacitance. Nat. Commun. 3, 736 (2012).
Carvalho-de-Souza, J. L. et al. Photosensitivity of neurons enabled by cell-targeted gold nanoparticles. Neuron 86, 207–217 (2015).
Bandara, H. M. & Burdette, S. C. Photoisomerization in different classes of azobenzene. Chem. Soc. Rev. 41, 1809–1825 (2012).
Hartley, G. S. The Cis-form of azobenzene. Nature 140, 281–281 (1937).
Rau, H. Spectroscopic properties of organic azo compounds. Angew. Chem. 12, 224–235 (1973).
Tang B. Z. & Qin A. Aggregation-Induced Emission: Fundamentals (Wiley, 2013).
Sierocki, P. et al. Photoisomerization of azobenzene derivatives in nanostructured silica. J. Phys. Chem. B 110, 24390–24398 (2006).
Fendler, J. H. Surfactant vesicles as membrane mimetic agents: characterization and utilization. Acc. Chem. Res. 13, 7–13 (1980).
Head, B. P., Patel, H. H. & Insel, P. A. Interaction of membrane/lipid rafts with the cytoskeleton: impact on signaling and function: membrane/lipid rafts, mediators of cytoskeletal arrangement and cell signaling. Biochim. Biophys. Acta 1838, 532–545 (2014).
Fruscione, F. et al. PRRT2 controls neuronal excitability by negatively modulating Na+ channel 1.2/1.6 activity. Brain 141, 1000–1016 (2018).
Thalhammer, A. et al. Alternative splicing of P/Q-type Ca2+ channels shapes presynaptic plasticity. Cell Rep. 20, 333–343 (2017).
Zhang, Q. & Bazuin, C. G. Liquid crystallinity and other properties in complexes of cationic azo-containing surfactomesogens with poly(styrenesulfonate). Macromolecules 42, 4775–4786 (2009).
Peddie, V., Anderson, J., Harvey, J. E., Smith, G. J. & Kay, A. Synthesis and solution aggregation studies of a suite of mixed neutral and zwitterionic chromophores for second-order nonlinear optics. J. Org. Chem. 79, 10153–10169 (2014).
Allen, J. A., Halverson-Tamboli, R. A. & Rasenick, M. M. Lipid raft microdomains and neurotransmitter signalling. Nat. Rev. Neurosci. 8, 128–140 (2007).
Frank, J. A., Franquelim, H. G., Schwille, P. & Trauner, D. Optical control of lipid rafts with photoswitchable ceramides. J. Am. Chem. Soc. 138, 12981–12986 (2016).
Hodgkin, Al & Huxley, A. F. A quantitative description of membrane current and its application to conduction and excitation in nerve. J. Physiol. 117, 500–544 (1952).
Tremere, L. A., Pinaud, R., Irwin, R. P. & Allen, C. N. Postinhibitory rebound spikes are modulated by the history of membrane hyperpolarization in the SCN. Eur. J. Neurosci. 28, 1127–1135 (2008).
Jo, S., Kim, T., Iyer, V. G. & Im, W. CHARMM-GUI: a Web-based graphical user interface for CHARMM. J. Comput. Chem. 29, 1859–1865 (2008).
Phillips, J. C. et al. Scalable molecular dynamics with NAMD +. J. Comput. Chem. 26, 1781–1802 (2005).
Huang, J. & MacKerell, A. D. Jr. CHARMM36 all-atom additive protein force field: validation based on comparison to NMR data. J. Comput. Chem. 34, 2135–2145 (2013).
Bennett, W. F. & Tieleman, D. P. Molecular simulation of rapid translocation of cholesterol, diacylglycerol, and ceramide in model raft and nonraft membranes. J. Lipid Res. 53, 421–429 (2012).
Maragliano, L. & Vanden-Eijnden, E. A temperature accelerated method for sampling free energy and determining reaction pathways in rare events simulations. Chem. Phys. Lett. 426, 168–175 (2006).
Dalla Serra, M. & Menestrina, G. Liposomes in study of pore-forming toxins. Meth Enzymol. 372, 99–124 (2003).
Dalla Serra, M. & Menestrina, G. Characterization of molecular properties of pore-forming toxins with planar lipid bilayers. Meth Mol. Biol. 145, 171–188 (2000).
Schmitt, B. M. & Koepsell, H. An improved method for real-time monitoring of membrane capacitance in Xenopus laevis oocytes. Biophys. J. 82, 1345–1357 (2002).
Pusch, M. & Neher, E. Rates of diffusional exchange between small cells and a measuring patch pipette. Pflug. Arch. 411, 204–211 (1988).
Gillis, K. D. Admittance-based measurement of membrane capacitance using the EPC-9 patch-clamp amplifier. Pflug. Arch. 439, 655–664 (2000).
We thank F. Fruscione and F. Zara (Giannina Gaslini Institute, Genova, Italy) for help in preparing iPSC-derived human neurons; P. Bianchini, M. Oneto and M. Scotto (Center for Nanoscopy and Nikon Imaging Center, Istituto Italiano di Tecnologia, Genova, Italy); L. Cingolani (Center for Synaptic Neuroscience and Technology, Istituto Italiano di Tecnologia, Genova, Italy) for providing the ChETA-encoding lentiviral vectors; A. Mehilli and G. Mantero (Center for Synaptic Neuroscience and Technology, Istituto Italiano di Tecnologia, Genova, Italy) for precious help in primary cultures and in vivo elelectrophysiology, respectively. This work was supported by the Italian Ministry of Health (project RF-2013-02358313 to G.P., G.L. and F.B.) and Istituto Italiano di Tecnologia (pre-startup project to G.L. and F.B.). The support of the Ra.Mo. Foundation (Milano, Italy), Fondazione 13 Marzo (Parma, Italy), Rare Partners srl (Milano, Italy) and Fondazione Cariplo (project 2018-0505) to G.L. and F.B. is also acknowledged.
The authors declare no competing interests.
Peer review information Nature Nanotechnology thanks Or Shemesh, Joao L. Carvalho-de-Souza and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
DiFrancesco, M.L., Lodola, F., Colombo, E. et al. Neuronal firing modulation by a membrane-targeted photoswitch. Nat. Nanotechnol. (2020). https://doi.org/10.1038/s41565-019-0632-6