Ifat

BABA GHULAM SHAH BADSHAH UNIVERSITY
COLLEGE OF ENGINEERING AND TECHNOLOGY
SEMINAR TOPIC
OPTOGENETICS
PRESENTED BY :
CO-ORDINATOR
IFAT RASHEED
MR.ARSHID BHAT
06-ECE-11
CONTENTS
INTRODUCTION
HISTORY
NEED OF OPTOGENETICS
COMPONENTS
DESCRIPTION
STATUS OF THE FIELD
RESEARCH NEEDS,OPPORTUNITIES AND CHALLENGES
EXPECTED OUTCOME AND BENEFITS
APPLICATIONS
ADVANTAGES
DISADVANTAGES
INTRODUCTION
Optogenetics is derived from a Greek word optos meaning visible.
It uses light to control neurons which have been genetically

sensitised to light.
It is a neuromodulation technique employed in neuroscience that uses

a combination of techniques from optics and genetics to control and
monitor the activities of individual neurons in living tissueeven
within freely-moving animalsand to precisely measure the effects
of those manipulations in real-time. .
It includes the discovery and insertion into cells of genes that confer
light responsiveness; it also includes the associated technologies for
delivering light deep into organisms as complex as freely moving
mammals, for targeting light-sensitivity to cells of interest, and for
assessing specific readouts, or effects, of this optical control.
What excites neuroscientists about optogenetics is control over

defined events within defined cell types at defined timesa level of
precision that is most likely crucial to biological understanding even
beyond neuroscience
CONTD
Optogenetics was established with the perspective of new approaches

on basic science and on the gene therapy of neural diseases such as
Parkinson disease,neuro psychiatric diseases and malfunctions in
vision.
Optical stimulation can be achieved by using caged compounds, e.g.

caged ATP,caged Glutamate,whereby the substrates for depolarizing
ion channels are delivered to membranes and activated by pulses of
UV- light to the chemical photolabile cage in the micro and milli
second scale.
Therefore, optogenetics is the combination of genetics and optics to

control well- defined events within specific cells of living tissue.
HISTORY
The far-fetched possibility of
using
light for
selectively controlling precise neural activity(action
potential) patterns within sub types of cells in the brain
was articulated by Francis Crickin .
In his Kuffler lectures at the University of California in
San Diego in 1999 .
An early use of light to activate neurons was carried out
by Richard Fork and later Rafael Yuste.
Recently in 2010, optogenetics was chosen as the
Method of the Year across all fields of science and
engineering by the interdisciplinary research journal-Nature Methods.
Three-gene
phototransduc
tion cascade
used to
activate cells
Discovery
and study
of opsins
1970s
Collaboration
between
Nagel, and
Deisseroth
(and Boyden)
Halorhodopsin
used for neural
silencing
Paper
published by
Chow et al,
(2010) using
archaerhodops
in which
completely
shuts down
the cell.
1999
Halorhodopsin
and neural
chloride levels
Paper on
channelrhodo
psin published
by Georg
Nagel et al.
(2003)
First published
paper using
optogenetics
(Boyden et al.,
2005) on
cultured
z
hippocampal
neurons,
followed by
papers from
several other
labs
Papers
published
using OptoXR,
light activated
GPCRs which
modulate
intracellular
signalling
(Aiden,2009)
and use in live
primates (Han
et al. 2009)
Use of
nanoparticles
and magnetic
pulses to
activate
specific cell
types without
such invasive
measures
(Palle Lab?)
NEED OF OPTOGENETICS
Optical
stimulation of electrically
excitable cells is superior to
classical
activation
by
microelectrodes .
Optogenetics
is known for the

high spatial and temporal
resolution that it provides in
altering the activity of specific
types of neurons to control a
subject's behavior
COMPONENTS
The
key reagents used in optogenetics are light

sensitive proteins .
Light-responsive proteins are allowing scientists to
turn neurons on or off selectively with
unprecedented precision.
Spatially
precise neuronal control achieved using

optogenetic actuators like channelrhodopsin,
halorhodopsin and archaerhodopsin while
temporally-precise recordings can be made with the
help of optogenetic sensors for calcium, chloride or
membrane voltage.
DESCRIPTION
CONTD
STATUS OF THE FIELD
After the application of ChR2 to neural cells ,the research field on

optogenetics developed extremely fast.
Major progress has been made on the transfection methods for

targeting different cell types in living animals, which allowed the
manipulation of ensembles of cells by light.
Genetically adapted to be expressed in the mammalian brain by viral

vectors, the microbial rhodopsins do the work with high precision .
Alternatively transfection has been achieved by electroporation

,however the virus approach appears to be more effective and has
biomedical perspective .
Recent work on cultured cells and on living mammals deals with

mapping of the brain and cellular mapping of neurons.
The results on the above mentioned topics prove that the optogenetic
approach is superior compared to the conventional electrophysiology.
RESEARCH NEEDS,OPPORTUNITIES AND

CHALLENGES:
Although
in the young field of optogenetics

spectacular results have been obtained,
many improvements are necessary with
respect to:
1. Transfection method
2. Optogenetic Tools and
3. Development of appropriate light sources for
studies in the brain or the retina of
mammals.
EXPECTED OUTCOME AND BENEFITS

The optogenetic approach offers for the future an
enormous potential for basic research, because nerve
excitation and silencing can be performed simply by light
with high precision in a reversible manner. Therefore,
research is focussed on neurological diseases, because all
the advantages of the light stimulation contain a
promising potential for gene therapy as described below:
1. Mapping of the brain and behaviour:

Immediately after having demonstrated that ChR2 can
be used for remote control of neurons, many laboratories
started projects for the brain in living animals. Excellent
work by various groups shows that the application of the
optogenetic methods opens the door in the near future
for more detailed studies, which have not been possible
with the traditional electrical and optical methods.
CONTD
2. Recovery of vision:
Experiments on photoreceptor deficient mice have shown, that light
evokes potentials in the visual cortex after the transduction of the
bipolar cells with ChR2 in the retina. This indicates that the retina of
the animals regain photo sensitivity.
3. Parkinsons disease and epilepsy:
Besides the application of drugs Parkinson's disease can be treated
by deep brain stimulation(DBS).Optogenetics offers great
opportunities for basic research as already has been demonstrated
by many laboratories worldwide.The biomedical applications,
however, hold unpredictable challenges and risks .
CONTD
4. Cell culture, Network
analysis:
The optogenetics
method provides new
opportunities to analyse
neural networks. This
can be achieved by
growing cultured nerve
cells on micro and nano
patterned substrates.
APPLICATIONS
Optogenetic activation and/or silencing has been used at least in

the following regions:
1. Amygdala:
Optogenetic approaches have been used to map neural circuits in
the amygdala that contribute to fear conditioning.
2. Nucleus accumbens:
The few cholinergic neurons present in the nucleus accumbens
may prove viable targets for pharmacotherapy in the treatment
of cocaine dependence.
3.
Prefrontal Cortex:
In vivo and vitro recordings expressing pyramidal neurons within
the prefrontal cortex demonstrated high fidelity action potential
output with short pulses of blue light at 20 Hertz.
CONTD
4.
Atrial Fibrillation: On atrial

cardiomyocytes was used as atrial
fibrillation to end spiral wave
arrhythmias zwith light. This
method is still in the
development stage.
DISADVANTAGES
Optogenetic studies provide many advantages compared to laser

based systems including :Illumination homogeneity and
stability,opsin selectivity,light switching,and
versatility.however,lasers do present some significant
disadvantages in performance and usability for optogenetics
studies including:
1.Mode Hopping(Diode lasers)

2.Back Reflection(Diode Lasers)
3.Speckle noise
4.Safety Tissues
5.Not suitable for wide field fluorescence microscopy
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BABA GHULAM SHAH BADSHAH UNIVERSITY

COLLEGE OF ENGINEERING AND TECHNOLOGY

STATUS OF THE FIELD

RESEARCH NEEDS,OPPORTUNITIES AND CHALLENGES

EXPECTED OUTCOME AND BENEFITS

Optogenetics is derived from a Greek word optos meaning visible.

It uses light to control neurons which have been genetically

It is a neuromodulation technique employed in neuroscience that uses

What excites neuroscientists about optogenetics is control over

Optogenetics was established with the perspective of new approaches

Optical stimulation can be achieved by using caged compounds, e.g.

Therefore, optogenetics is the combination of genetics and optics to

is known for the

key reagents used in optogenetics are light

precise neuronal control achieved using

STATUS OF THE FIELD

After the application of ChR2 to neural cells ,the research field on

Major progress has been made on the transfection methods for

Genetically adapted to be expressed in the mammalian brain by viral

Alternatively transfection has been achieved by electroporation

Recent work on cultured cells and on living mammals deals with

RESEARCH NEEDS,OPPORTUNITIES AND

in the young field of optogenetics

EXPECTED OUTCOME AND BENEFITS

1. Mapping of the brain and behaviour:

Optogenetic activation and/or silencing has been used at least in

Atrial Fibrillation: On atrial

Optogenetic studies provide many advantages compared to laser

1.Mode Hopping(Diode lasers)

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