General introduction

into optogenetics

http://www.cairn-research.co.uk/applications/optogenetics-the-led-light-switch-neurons

Represented by Cuong, Pham Van

Outline
1. What is optogenetics?
2. Timeline of development of
optogenetics
3. Process of optogenetics
4. Application of optogenetics
5. Beyond the neuroscience

Outline
1. What is optogenetics?
2. Timeline of development of
optogenetics
3. Process of optogenetics
4. Application of optogenetics
5. Beyond the neuroscience

What is optogenetics?
Optogenetics is the combination of
genetic
and optical methods to achieve gain or loss
of function of well-defined events in
specific
cells of living tissue
Karl Deisseroth (2011)

Outline
1. What is optogenetics?
2. Timeline of development of
optogenetics
3. Process of optogenetics
4. Application of optogenetics
5. Beyond the neuroscience

Timeline of development of optogenetics

Functional unit
of nervous
system: Neuron

Early
20th
centur
y

197
0s

2002 2005

Functional unit of nervous system: Neuronsignal transmitter

By changes in membrane potential due to opening/closing
specific ion pump channel
2 type of changes: depolarizing (excitatory stimulus)
hyperpolarization (inhibitory stimulus)

http://classes.midlandstech.edu/carterp/Courses/bio210/chap11/lecture

Timeline of development of optogenetics

Functional unit
of nervous
Idea of
system: Neuron
neural
control by
light
Early
20th
centur
y

197
0s

2002 2005

Idea of neural control by light

Challenge: how to control one type of
cells in brain while not influence
others?
Inappropriate factors: electorde (no
precisely target), drug ( broaden
effect, too much slowly)
Light might have properties as a
control tool

Thinking about brain, Francis Crick (197

Timeline of development of optogenetics

Functional unit
of nervous
Idea of
system: Neuron
neural
control by
light
Early
20th
centur
y

197
0s

Discovery of opsins:
Bacteriorhodopsin
Halorhodopsin

2002 2005

Discovery of opsins:
visible-light-activated ion pumps

Microbial ion pump activated by visible light

(1971)
Bacteriorhodopsin:
Extrude proton to
create proton gradient
for production of ATP
(1977) Halorhodopsin
(NpHR)
Transport chloride into
cytoplasm to maitain
osmotic balance
Oesterhelt and Stoeckenius, 1971
MatsunoYagi and Mukohata, 1977

10.1016/B978-0-444-59426-6.00001-X

Timeline of development of optogenetics

Functional unit
of nervous
Idea of
system: Neuron
neural
control by
light
Early
20th
centur
y

197
0s

Discovery of opsins:
Bacteriorhodopsin
Halorhodopsin

2002 2005

Channelrodopsins
described

Channelrodopsins
From unicellular photosynthetic
organism
Used for light orientation

2 type:
ChR1 (permeable to
photon)

ChR2

(permeable to
other cations)

Nagel et al.,

10.1016/B978-0-444-59426-6.00001-X

Timeline of development of optogenetics

Functional unit
of nervous
Idea of
system: Neuron
neural
control by
light
Early
20th
centur
y

197
0s

Discovery of opsins:
Bacteriorhodopsin
Halorhodopsin

First application of opsins

to neurosciense

2002 2005

Channelrodopsins
described

First application of opsins to neurosciense

By Boyden et al.,
2005
Successful
expression of ChR2 in
cultured mammalian
hippocampal neurons
ChR2 activation by
light was enough to
produce action
potential
http://www.cell.com/cms/attachment/
2007954326/2030561053/gr1.jpg

Outline
1. What is optogenetics?
2. Timeline of development of
optogenetics
3. Process of optogenetics
4. Application of optogenetics
5. Beyond the neuroscience

Process of optogenetics
1. Choosing
opsins of
interest
2. Using viral
carrier to
express
opsins
3. Delivering
light into
brain
Pama et al., 2013

http://optogenetics.weebly.com/why--how.html

Process of optogenetics
1. Choosing
opsins of
interest
2. Using viral
carrier to
express
opsins
3. Delivering
light into
brain
Pama et al., 2013

http://optogenetics.weebly.com/why--how.html

Choosing opsin of interest

Excitatory by
ChR2 with blue
light

Inhibitory by
NpHR with yellow
light
Pama et al., 2013

Process of optogenetics
1. Choosing
opsins of
interest
2. Using viral
carrier to
express
opsins
3. Delivering
light into
brain
Pama et al., 2013

http://optogenetics.weebly.com/why--how.html

Using viral carrier to express opsin

Lenti and adeno-associated
virus (AAV) is commonly used
high expression level over long time period, no
adverse effects

Site-specific enhance by celltype-specific-promoter or Cre

recombinase-dependent
expression system

(Pama et al., 2013; Yizhar et al., 2011)

Process of optogenetics
1. Choosing
opsins of
interest
2. Using viral
carrier to
express
opsins
3. Delivering
light into
brain
Pama et al., 2013

http://optogenetics.weebly.com/why--how.html

Delivering light into brain

Using chronically
implanted cannula
(ng cy ghp c nh)

(affixed to skull)
Pama et al., 2013

Attached optical
fiber: to delivering
laser light directly
into brain
http://www.nytimes.com/2011/05/17/scien
ce/17optics.html?_r=1

Outline
1. What is optogenetics?
2. Timeline of development of
optogenetics
3. Process of optogenetics
4. Application of optogenetics
5. Beyond the neuroscience

Application of optogenetics
Advantage: fast (in millisecond scale)
and precision (cell type- specific)
Example:
In neuromodulation research
Using optogenesis to show how symptoms of
Parkinsons disease can be either aggravated or
improved (Kravitz et al., 2010)
In sleep research
Examining boundaries between sleep and wakefulness
by applying optogenetics to control neural circuit
Karl Deisseroth, 2010
Pama et al., 2013

Outline
1. What is optogenetics?
2. Timeline of development of
optogenetics
3. Process of optogenetics
4. Application of optogenetics
5. Beyond the neuroscience

Beyond the neuroscience

Appropriate for the target tissue of
choice
With regard to electrical or
biochemical effector function, speed
and other properties
Example: Control biochemical,
electrical drive in culturing stem cell
Karl Deisseroth, 2010

Thanks for attending and

listening!