engineering

Switching on the Light Bulb

technology

Lighting the Way in

Neural Circuit
Dynamics and Depression
by Caitlyn McCullough
Photo Credit: Karl Deisseroth

W
e are all familiar with the proverbial light bulb
inside our head that flickers with the ebb and
flow of ideas. Dr. Karl Deisseroth, Assistant
Professor in Stanford’s Department of Bioengineering and
the Department of Psychiatry and Behavioral Sciences, has
stepped beyond the figurative, developing a tool that may
one day lead to the use of an LED implanted in the brain to
stimulate neurons in the treatment of neurological diseases
and psychiatric disorders.
The spotlight of his work is the light-gated cation-selective
membrane channel recently discovered in green algae called
channelrhodopsin-2 (ChR2). Although photostimulation
of neurons in direct clinical intervention is a distant goal
with many challenges, the Deisseroth lab has demonstrated
millisecond-timescale optical control of neurons in a
genetically targetable and temporally precise manner using
ChR2. Such unprecedented selectivity and precision may
In the Light
The gene was not initially known to neuroscience, having
been identified in Chlamydomonas reinhardtii in 2001 by
algae biologists in Germany. ChR2 is a seven-transmembrane
protein with a light-sensitive cofactor called “all-trans-
retinal” (ATR) bound at the core. Blue light isomerizes the
ATR core, resulting in an open pore that allows selective
cations to diffuse into the cell, causing depolarization. If the
threshold potential is reached, voltage-gated ion channels
open, initiating an action potential and the passage of an
electrical signal across the synapse to another neuron.
Could this directly light-gated ion channel be the means of
optical control that neuroscientists have been in search of
for decades?
Deisseroth reached out to
the algae experts (Nagel et al.
in Germany), hypothesizing

Photo Credit: Karl Deisseroth

revolutionize how neuroscientists and bioengineers view that this gene could be
neural circuit dynamics and neurological diseases. brought to neuroscience and
bring great advances to the
In the Dark study of neural circuits. With
The brain consists of a dense architecture of heterogeneous such great gains, equal risk
neural cell subtypes, each presumed to play a specific role in was involved. Deisseroth and
gating of information and electrical rhythms. It is estimated his colleagues undertook this
that hundreds of neural subtypes exist, and understanding high risk project, investing
their respective contributions to neural processing is important time and effort, unsure if the Karl Deisseroth is an assistant
in elucidating fundamental concepts of basic neuroscience, channel would work without professor in both the Department
identifying aberrations in neurological diseases and psychiatric the addition of exogenous of Bioengineering and the
disorders, and developing interventions for treatment of ATR. “If you pooled all the Department of Psychiatry at
Stanford University.
disease. To probe neural circuits for subtype involvement, a scientists involved, 99 out of
means of selective excitation or inhibition of neural subtypes 100 would have said, ‘No way,
in a temporally precise, accurate, and physiologically relevant it’s not going to work,’” without addition of the cofactor; this
manner within an intact circuit is required. had been the stumbling block for previous photostimulation
Current methods of probing neural circuits, through the attempts. Fortunately, the efforts have paid off. Retinoids
use of electrodes or optical methods, have not achieved this present in mammalian systems allow for ChR2 to function
combination of spatial selectivity and temporally precision. properly. “We had a prepared mind and were able to bring
Optical methods such as light-mediated uncaging of signaling [ChR2] to neurons first - (published in Nature Neuroscience
molecules, chemically modified ion channels and receptors, in 2005),” Deisseroth comments.
and the use of naturally occurring photosensitive proteins,
have thus far been complicated by the difficult administration In the Lab
of exogenous cofactors or by requiring downstream The Deisseroth lab has demonstrated noninvasive,
machinery. Despite these challenges with optical control genetically targeted, millisecond-timescale activation in
of neurons, the advantages of photostimulation have kept both in vitro cultured rat neurons and in vivo hippocampal
many, including Deisseroth and his colleagues, “carefully regions of mice. Genetic modification of neurons is achieved
watching this literature for years.” using lentiviral vectors engineered to encode for the ChR2-
42 stanford scientific

Deisseroth has stepped beyond the figurative, developing a tool that may
one day lead to the use of an LED implanted in the brain to stimulate neurons
in the treatment of neurological diseases and psychiatric disorders.
yellow fluorescent protein (YFP) fusion protein, as well
as a cell-specific promoter and components to improve
longevity of expression (Fig 1a). Using a commonly found
promoter, high levels of ChR2-YFP are expressed in the
mouse hippocampus (Fig 1b). Excitation of in vitro neuron
cultures with blue light induces rapid inward current (Fig
1c) and depolarization (Fig 1d). Five superimposed traces
of a spike train indicate the temporal precision and accuracy
of stimulation (Fig 1e). Depending upon the length of the
light impulse, subthreshold potentials and action potentials
can both be generated. Stimulation of in vivo neurons with
ChR2 can also be achieved using an implanted fiberoptic
cable. The success of ChR2 in mammalian systems poses
great promise for basic neuroscience and bioengineering.
In the Future
Selective excitation using ChR2 has widespread
applications, including basic neuroscience concepts,
Photo Credit: Karl Deisseroth
Channelrhospsin-2 (ChR2) is shown to be a valuable tool. A lentiviral
vector is used for delivery of ChR2 (a), ChR2-YFP is expressed in mouse
hippocampus dentate granule cells (b). Blue light induces inward current
(c), depolarization (d), and temporally precise, accurate, and sustainable
spike trains (e) in mouse neuron without exogenous cofactor.
layout design: Natasha Prats
engineering
+
technology
guiding stem cell differentiation, ion channel drug discovery,
and treatment of depression. “ChR2 allows us to do all optical
interrogation of neural cells. There are existing [optical]
voltage and calcium sensors that allow us to observe the
activity of the cell; now we can also optically control them.
This opens the door to high throughput all optical screening,”
comments Deisseroth.
It is estimated that ion channel drug discovery
(presently done using the patch-clamp method, a physical
interaction of a tiny pipette with an ion channel) could be
accelerated by a factor of 1000 or more with this method.
Neurological diseases and psychiatric disorders could be
better understood, as well as improvement of treatments for
such aberrations. Current treatment of Parkinson’s disease,
epilepsy, and increasingly, depression, involve deep brain
stimulation with an electrode. Excitatory cells, inhibitory
cells, and other cells in the region are activated by the patch-
clamp method, creating a source of associated side effects
and decreased efficacy. By using ChR2, the specific roles
of neural subtypes in normal and abnormal functions can
be determined. Excitation of targeted neural populations
involved in the treatment of disease or psychiatric disorders
can then be achieved using ChR2.
Deisseroth imagines the use of an inductively controlled
implanted wireless LED to activate ChR2 in genetically
targeted neural cells to interfere with electrical rhythms
involved in depression. By no means is this a trivial task;
light and gene delivery are two inherent challenges that need
to be further investigated.
Deisseroth continues to make advances in the use of ChR2
with other photostimulation techniques in order to modulate
and observe neural activity in an all optical system. He is
opening channels, literally, for neuroscientists and bioengineers
to understand neural circuit dynamics, neurological diseases
and psychiatric disorders, and ways in which intervention may
one day bring light to the brain. “[ChR2] is a very promising
addition to our tool belt,” says Deisseroth. “It opens the door
to a whole new way of doing things and we’re excited to be
working on it here at Stanford.” S
CAITLYN MCCULLOUGH is a first-year graduate student in
Bioengineering. She enjoys racing bikes, and is a member of the
Stanford Cycling Team and Team Tibco.
To Learn More
Visit the Department website of Dr. Karl Deisseroth:
http://www.stanford.edu/groups/dlab
Read Zhang F,Wang LP, Boyden ES, Deisseroth K. Channelrhodopsin-
2 and optical control of excitable cells. Nat Methods. 2006 Oct;
3(10):785-92.
Read Boyden ES, Zhang F, Bamberg E, Nagel G, Deisseroth K.
Millisecond-timescale, genetically targeted optical control of
neural activity. Nat Neurosci. 2005 Sep;8(9):1263-8.
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