CH391L/S13/Optogenetics

Introduction to Optogenetics

"Before we can find the answers, we need the power to ask new questions." -Karl Deisseroth

In essence, optogenetics is a neural modulation technique used to control neurons in vitro for the purpose of affecting the physiology of neural circuits and ultimately behavior of the studied organism. However, in recent studies, optogenetic techniques have been used to modify nonneuronal tissues, such as cardiac tissue and beta cells, willfully controlling the respective cell-specific roles. Although the implications of optogenetics seem like a panacea for many genetic diseases, much of the field is new; in terms of therapeutics, research is regretfully far behind. Currently, optogenetic techniques are attempted mostly on rodent specimens since primate studies lack profound electrophysical and behavior effects. Optogenetic is widely associated with neuroscience research- sometimes thought as the synergy between neuroscience and synthetic biology. Current optogenetically-related research aims to ascertain brain function of multiple neural circuits, but future endeavors include gene therapy for neurodegenerative diseases and neuroprosthetics.

The basis is optogentics is quite simple. The neuron in question is spliced with a specific opsin gene carried by viral vector, usually a modified lentivirus. Subsequently, the encoded opsin attached to the cell membrane. Opsin are photosensitive G protein receptors or ion channels that are induced by a specific wavelength of visible light via fiber optic cable or optrode. In turn, the opsin undergoes a conformational change, eliciting a change in membrane potential. For neuronal cells, changes in membrane potential give rise to action potentials. The strength of the depolarizing current (incoming positive charged ions) is encoded in the frequency of the action potentials generated. Furthermore, synapses (gap junction between neurons) can conduct spatial or temporal summation. Even neuronal cells can be silenced with hyperpolarizing current (incoming negative or outgoing positive charged ions). In short, optogenetics provides neuroscientists with an “on/off” switch for targeted neurons.

History

"The brain is a world consisting of a number of unexplored continents and great stretches of unknown territory." - Santiago Ramón y Cajal

Year	Event
1971	Bacteriorhodopsin (yellow and green)
1977	Halorhodopsin (yellow)
1999	Francis Crick's Declaration
2002	ChR1 (blue), Zemelman and Miesenbock chARGed optogenetic system
2003	ChR2 (blue)
2005	Millisecond-timescale, genetically targeted optical control of neural activity
2006	"Optogenetics" coined
2008	VChR1 (yellow and possibly green)

The story behind optogenetics is a story of convergence of two seemingly unrelated fields: microbial microbiology and neuroscience. In 1971, Walther Stoeckenius and Dieter Oesterhelt discovered the transmembrane protein bacteriorhodopsin (BR) that is capable of moving ions in the presence of light. BR is highly expressed in haloarcheal membranes while a similar transmembrane protein proteorhodopsin (PR) is expressed in eubacteria. In 1977, Matsuno-Yagi and Mukohata described the ion pump channel halorhodopsin which displaces extracellular chloride ions in to the cell. As these discoveries were happening in the background, a growing desire to effectively control the neurons was ever –prevalent. During this period, neuroscientist adopted the idea that cognitive processes were distributed in various localizations of the cortex. This fact evident when lesions in specific areas would lead to loss of a specific process. Even fMRIs and PET solidified this point, but, even with these high-tech instruments, the processes themselves could not fully be understood. This would require an in detail analysis of the neural circuits that make up the localized cortical space. This would entail the use of electrodes in order to stimulate specific cells to generate action potentials. Electrode analysis and other biochemical methods provided much of the insight that we currently have on neuroscience, but this systematic approach is inherently flawed. Even though the electrodes are made extremely small in order to excite a particular cell, the electric charge permeates the cerebral space so that other close neurons are also stimulated. Such a problem defeats the purpose of ceteris paribus, since we are unable to determine whether the cell in question elicited the desired response in the first place. Moreover, to conduct a loss of function experiment, the neural tissue is question needs to be destroyed, or lesioned, but, once the tissue is destroyed, there are no methods to restored said tissue, further complicating neural circuit research. This lack of cell specificity and cellular controls encouraged neuroscientist to find alternative methods. In the 1990s, some scientist became infatuated with the idea that opsins may be the key; however, in the most part, this idea was considered too “far-fetched” to allocate resources, time, and money. In 1999 at the University of California in San Diego, Francis Crick commented that neuroscience faces the major challenge of controlling one type of cell while leaving the others unaffected. Crick even suggested that light could serve as a control tool. In 2002, Boris Zemelman, currently a UT associate professor, and Gero Miesenbock conducted the earliest genetically targeted method with rhodopsin photoreceptors in Drosophila. Also in 2002, channelrhodopsin (ChR1) was identified by Georg Nagel to contain a wider cation conductance (Na and K ions). In 2003, Channelrhodopsin-2 (ChR2) was later discovered in the same organism Chlamydomonas reinhardtii which conducts cations more than twofold greater than those of ChR1. ChR2 became the key to excitatory neuron transmission. In 2005, Karl Deisseroth, Edward S. Boyden, and Feng Zhang created a robust ChR2 optogenetic system cultured in mammalian neurons for precisely timed light driven action potentials. The term “optogenetics” was coined in 2006. In brief, the opsin genes discovered in various microbes were spliced in the neuron in order to elicit desired neural activity.

Applications of Optogenetics

IGEM Take-home Message

http://2012.igem.org/Team:Washington/Optogenetics http://2006.igem.org/wiki/index.php/University_of_Texas_2006 http://2006.igem.org/wiki/index.php/UT_Austin_2005 http://2004.igem.org/austin.cgi

References

1. Mei Y and Zhang F. Molecular tools and approaches for optogenetics. Biol Psychiatry. 2012 Jun 15;71(12):1033-8. DOI:10.1016/j.biopsych.2012.02.019 |
2. Yizhar O, Fenno L, Zhang F, Hegemann P, and Diesseroth K. Microbial opsins: a family of single-component tools for optical control of neural activity. Cold Spring Harb Protoc. 2011 Mar 1;2011(3):top102. DOI:10.1101/pdb.top102 |
3. Zemelman BV, Lee GA, Ng M, and Miesenböck G. Selective photostimulation of genetically chARGed neurons. Neuron. 2002 Jan 3;33(1):15-22. DOI:10.1016/s0896-6273(01)00574-8 |
4. LaLumiere RT. A new technique for controlling the brain: optogenetics and its potential for use in research and the clinic. Brain Stimul. 2011 Jan;4(1):1-6. DOI:10.1016/j.brs.2010.09.009 |
5. Doroudchi MM, Greenberg KP, Liu J, Silka KA, Boyden ES, Lockridge JA, Arman AC, Janani R, Boye SE, Boye SL, Gordon GM, Matteo BC, Sampath AP, Hauswirth WW, and Horsager A. Virally delivered channelrhodopsin-2 safely and effectively restores visual function in multiple mouse models of blindness. Mol Ther. 2011 Jul;19(7):1220-9. DOI:10.1038/mt.2011.69 |
6. Boyden ES, Zhang F, Bamberg E, Nagel G, and Deisseroth K. Millisecond-timescale, genetically targeted optical control of neural activity. Nat Neurosci. 2005 Sep;8(9):1263-8. DOI:10.1038/nn1525 |