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Optogenetics: What is it? What can it do? What to expect in the future? Build an understanding from the fundamentals of life science.

 

Optogenetics: What is it? What can it do? What to expect in the future? Build an understanding from the fundamentals of life science.

This article is aimed to help you understand the new technology of Optogenetics from the fundamentals of biology so that you can build knowledge, rather than accumulate discrete facts. All explanations have been included under a heading, so feel free to skip the ones you are already familiar with. Some instances of how a conclusion was arrived at have also been discussed through experiments so that you can understand the inner workings of scientific laboratories. Happy learningJ

 

For original article, please visit: https://medium.com/@adachoudhry26/optogenetics-what-is-it-293f3fdded56

 

 

 

 

 

 

 

 

 

 

Contents:

·      Crash course in Biology

1.   Cell: The building block of life

2.   Neuron : The building of Nervous System

3.   Functions of the Nervous System

A.  Sensory

B.   Integrative

C.   Motor

·      What is Optogenetics?

1.   Understanding Genetic Engineering

A.  What are genes?

B.   Do all cells (even neurons) have DNA (the carrier of genes)?

·      How Optogenetics works?

                                                      I.            Step 1: Modify the neuron

v Understand Photosynthesis

v Understand Action potential

                                                   II.            Step 2: Insert an optical Fibre into the brain of the animal to be studied

v Understand Optical Fibre

·      Let’s learn!

v Understanding electrical signaling in the brain

v How do neurons perceive time?

 

·      How the technology is being used today?

1.   For treating neurological disorders

§  Parkinson’s Disease

v Understanding Deep Brain Stimulation (DBS)

v Neural grafting

2.   For finding neurological cures for mental health disorders

§  Elevated rat maze experiment for anxiety

3.   As a memory modifying technique

·      Why use Optogenetics?

o  Story of the first road map to the brain

·      What the future holds?

§  Ethical considerations of it’s memory modifying potential

§  Refining cures of neurological disorders

·      Bibliography

 

 

 

Crash course in Biology

To understand the mechanism of optogenetics, let’s start from the very basics of biology.

Cell: The building block of life



Cell is the basic structural and functional unit of life. It comes from the Latin word cellula meaning small room. It is the fundamental structural unit because all organisms can be simplified on the basis of cells.



Think of cells like Lego blocks that on being attached together form a structure (organism). Some organisms are unicellular which means that they have only one cell which performs all the basic functions important for sustaining life like metabolism (Metabolism is the process by which your body converts what you eat and drink into energy (2)) , growth (development towards functional efficiency and gradual increase in size) and reproduction (production of new organisms from existing organisms). Due to its self sustaining nature, cell is known as the fundamental functional unit of life. Some organisms are multicellular, which means that they have many cells in their body. The cells are structurally differentiated on the basis of their location which affects their function. Human beings are multicellular organisms.

Neuron: The Building block of Nervous System



In multicellular organisms, there is a type of cell known as the neuron which is the building block of the nervous system.

Image Source: cusabio.com

 The nervous system is the major controlling, regulatory, and communicating system in the body” (3) Any function that is performed by the body is orchestrated by the nervous system. It gives instructions to the organs through a network of neurons known as the nerves. “It is the center of all mental activity including thought, learning, and memory.  Together with the endocrine system, the nervous system is responsible for regulating and maintaining homeostasis. Through its receptors, the nervous system keeps us in touch with our environment, both external and internal.” (3)

·     



Functions of the nervous system

In vertebrates, the nervous system is formed of brain, spinal cord, nerves and ganglia. There are three basic functions of the nervous system:

·      Sensory (Input)

·      Integrative (Processing)

·      Motor (Output)

Thus, the functioning of the nervous system resembles the IPO cycle of a computer.



Sensory: Collecting information (input). As it is a communication system, it helps us to sense our surroundings by transmitting different types of messages from sense organs. It also receives messages from different organs so as to regulate internal functions and processes. This is known as Sensory input.

Integrative: The processing of input comes under the integrative function of the nervous system. The Sensory input is converted to electrical signals known as nerve impulses and transmitted to the brain. This input is brought together to form sensations (Meaning: a physical feeling. All physical experiences are a result of these sensations), thoughts (electro-chemical reactions in the brain. A single thought is a result of thousands of signals passing through the synapses [meeting point of two nerves] in the brain) or store the input in memory if it may be relevant for future use. Storing important data in the brain comes under learning. Based on the result of processing, decisions are made every moment.

Motor: On the basis of the decision taken, a response is sent to glands or muscles. This is the motor output.

A question that rose in my mind when learning about neurons was how do they communicate these messages to and fro from Sensory organs to Central Nervous system and then again to Motor organs?

The discovery of the medium used by neurons takes us back in time to the hustle and bustle of an Italian market in the midst of a lightning storm, when scientist Luigi Galvani saw that the legs of the frogs being sold were twitching. He concluded that electricity must be activating the nerves of the frog. Repeating the experiment in his laboratory he found out that he was right! So the scientific world learned that neurons communicated through electrical signals. We literally have electricity running through our body.

What is Optogenetics?

Now that we have learnt about the basics of nervous system, let’s understand about the upcoming technology of Optogenetics.



Opto- is a prefix meaning vision and Genetics refers to the study heredity or inheritance in organisms carried out by genes.


Optogenetics is a method for controlling the activity of a neuron by using light and genetic engineering. In this process, scientists shine light onto genetically modified neurons that are light sensitive which changes their signals. Neurons have two functions: they can either block or transmit the nerve impulse and shining light on them reverses their output. It is defined as an invasive neuromodulation technique.

·      Understanding genetic engineering

Genetic engineering  is defined as “The process of changing the information in the genetic code (the blueprints) of a living organism by adding or deleting information. Genetic engineering is sometimes called genetic modification.” (1). During the process of reproduction, the parent organism passes a set of codes and instructions to specify traits to the offspring in the form of genes.

 

·      What are genes?




The gene is the basic physical unit of inheritance. Genes are arranged, one after another, on structures called chromosomes. A chromosome contains a single, long DNA molecule, only a portion of which corresponds to a single gene. Humans have approximately 20,000 genes arranged on their chromosomes.” (4)

·      Do all cells have DNA (the carrier of genes)?

 

“Nearly every cell in a person’s body has the same DNA. Most DNA is located in the cell nucleus (where it is called nuclear DNA), but a small amount of DNA can also be found in the mitochondria (where it is called mitochondrial DNA or mtDNA). Mitochondria are structures within cells that convert the energy from food into a form that cells can use.” (5)



The DNA can replicate which is an important property as each cell needs to have the exact copy of the DNA present in the old cell. Thus, even neurons contain DNA which contain genes that can be modified to affect their performance.

How Optogenetics works?

To perform Optogenetics, scientists use two steps:

Step 1: Modify the neuron



Algae are aquatic organisms (most of them are unicellular that are capable of forming oxygen through photosynthesis. Photosynthesis is a process through which plants having the green pigment, chlorophyll, synthesize energy in the form of glucose using water and carbon dioxide in the presence of sunlight. Oxygen is formed as a by-product. As sunlight is crucial for the production of glucose and energy (organisms cannot function without energy), Algae contain a light sensitive eye spot which is a photo receptive organelle. Photo receptive means that it is activated on receiving photons (light particles) and organelle is a subcellular structure. This organelle contains the light sensitive pigment channelrhodopsin. “Channelrhodopsin allows algae to perceive water depth using light: the deeper they go, the less light there is. These proteins control ion channels, making them capable of generating an action potential in neurons.” (15)

 

Action potentials (those electrical impulses that send signals around your body) are nothing more than a temporary shift (from negative to positive) in the neuron’s membrane potential caused by ions suddenly flowing in and out of the neuron.” (16)

 



When blue light shines on the eyespot of an Algae, it sends out electrical signals that make flagella flap. Flagella is microscopic hair-like extension which is involved in the locomotion of a cell. Scientists clone the eyespot which is capable of sending electrical signals and genetically modify it in the neuron to be studied.


Step 2: Insert an optical Fibre into the brain of the animal to be studied.

An optical fiber is a flexible, transparent fiber made by drawing glass (silica) or plastic to a diameter slightly thicker than that of a human hair.” (5) Data is transmitted in these wires using light pulses. On flipping the switch on, scientists can target specific light sensitive neurons in the brain to study how they communicate with others. This way neurons can be studied in specific parts of the brain without affecting the functioning of other unrelated parts.


Let’s learn!

·      Understanding electrical signaling in the brain


Image source: Furman News – Furman University

Electrical signaling happens among the different and multiple neurons in the brain which determines the brain functions. The generated electrical signals are in the order of milliseconds. These signals are spatiotemporally encoded which means that they depend on space (location) and time.

·      How do neurons perceive time?

 In biological system, time is measured according to different rhythms like on the macro scale the circadian rhythm or the

sleep-wake cycle is measured in days or months. “At the organismal level, daily oscillations come about through coordination of tens of thousands of neurons situated in the suprachiasmatic nucleus (SCN).” (7) In the brain,   suprachiasmatic nuclei are two small, paired nuclei that are found in the hypothalamus.” (9)


Image source: National Institute of General Medical Sciences 

“Despite numbering only about 10,000 neurons on each side of the third ventricle, the SCN is our principal circadian clock, directing the daily cycles of behaviour and physiology that set the tempo of our lives.” (8)

Thus, time on the biological scale depends on rhythms or oscillations.

 Neurons are capable of telling time on the scale of tens to hundreds of milliseconds based on their firing dynamics. The molecular clock that is born from this, by virtue of controlling

cellular processes, controls the organization of the three spatial dimensions. On the other hand, in biological systems the fourth dimension, time, can be born from the three dimensions of space.” (7)

They are able to do this as they are located in intricate three-dimensional circuits in the brain. “Over time, the electrical signals form activity patterns of the brain. Actually, these patterns encode our thoughts, skills, feelings, and memories, and thereby control the brain function and then its resulting behavior. However, how this process is done is unknown.” (10)

Optical, instead of electrical,  stimulation allows the study of specific neural subsets of the brain that give rise to complex activities as neurons respond to exciter within milliseconds.

 

How the technology is being used today?

·      Treating neurodegenerative diseases

Simulation of how the neural circuits operate helps the scientists to understand brain activities and find treatments of neurodegenerative diseases. “Neurodegenerative diseases occur when nerve cells in the brain or peripheral nervous system lose function over time and ultimately die. Although treatments may help relieve some of the physical or mental symptoms associated with neurodegenerative diseases, there is currently no way to slow disease progression and no known cures.” (12)

·      Parkinson’s Disease

Parkinson’s is a neurodegenerative disease in which neurons in the brain degenerate or lose their function and ultimately die. It is a movement disorder which results in tremors in patients due to disorganized electrical signals in the areas of the brain that control movement. The exact cause of this disease is unknown,  however certain genes make people more susceptible to it as they age. This disease results in the death of dopamine releasing neurons in the brain. Current treatments include administering dopamine to patients and deep brain stimulation (DBS).


Image: DBS for Parkinson’s Disease, Source: Elite Ayurveda

Deep brain stimulation is a neurosurgical procedure in which doctors insert electrodes in the concerned circuits to block irregular signals. These methods have long lasting side effects. DBS is a time consuming process in which doctors have to ensure that electrodes do not interfere with unrelated neurons through a series of tests before and after the procedure to correctly locate the target cells.

Optical stimulation through Optogenetics allow to selectively target axons in specific regions to either drive or inhibit an array of distinct circuit elements. After locating the cause of the irregular signals, the cells can be replaced using neural grafting to host connectivity. “Neural grafting involves placing cells, either as a block of tissue or as a suspension of dissociated neuronal cells, into a predetermined area of the central nervous system (CNS).” (12). Thus neural transplantation has emerged as a possible treatment of Parkinson’s disease that can be successfully done through Optogenetics by identifying the target cells.

·      For finding neurological cures for mental health disorders

“Optogenetics has made it possible to determine which cells and connections across the brain are important in defining and assembling the different features of anxiety, including respiratory-rate changes and risk avoidance, into a distinct behavioral state.” (13) Thus, Optogenetics is helping in finding a neurological treatment of mental health disorders.

·      Elevated rat maze experiment for anxiety


In the laboratory of Kay M. Tye, experiments done on mice revealed that amygdala, part of the brain which regulates emotions, has two different neural connections – seeking pleasure and avoiding pain. These two tendencies describe a majority of our behaviour. In anxiety patients, avoiding pain is a marked behaviour and over time, this neural pathway is strengthened. In the experiment, when a mouse is kept at a corner of the maze, it seldom explores the rest of the surroundings as they might signal potential dangers. This is an example of avoidance of pain. When light is shown on the target neuron, it can either transmit or block the nerve impulse. In this experiment, the nerve impulses were blocked in the circuit which avoids pain. When the light was switched on, the mouse freely explored the rest of the maze. On switching the light off, the mouse retreated to the safety of its corner.

Image: The elevated plus maze. Measuring the time a rat spends exploring the maze versus the time spent in the safety of closed arms. Source: Conduct Science

Thus, Optogenetics offers a way to control the brain instantly, reversibly and without side effects.

·      As a memory modifying technique

Optogenetics provides even greater memory-modifying opportunities than have previously been assumed including selective and reversible erasure (repeated deactivation and reactivation) of a specific memory and the retrieval of forgotten (or suppressed) memories.” (11) This new avenue is an issue of neuroethics and debate of fundamental right and wrong, which is discussed in the section for future implications of the technology. Optogenetics could be used as a possible memory modifying technique (MMTs).

Why use Optogenetics?

Optogenetics was first discovered in 2005 and is now being used in laboratories across the world.

Any technique no matter how cool it sounds must have advantages to be utilized in scientific laboratories. After Galvani discovered that neurons communicate through electrical signals, scientists began experimenting on neural pathways by inserting electrodes into the brains of humans or animals (as humans have similar brain structures to certain animals).

·      Story of the first road map of the brain

The first brain mapping was done through electrical stimulation by Dr. Wilder Penfield, a brain surgeon who was working with epilepsy patients. Epilepsy is “a disorder in which nerve cell activity in the brain is disturbed, causing seizures.” (6)

In extreme cases, surgery had to be performed. Dr. Penfield wanted to know which parts of the brain were vital so he could avoid operating on them. He lowered electrodes into the motor areas of the brain which control movement and passed a signal to find that one area resulted in twitching of the finger while one caused twitching of the toe. With this experiment, he was able to learn which part of the brain controlled different parts of the body. This resulted in the first brain map, a road map which could tell us how the brain communicated and controlled the body. As we have just seen, studies on the brain required putting electrodes in the brain which could result in permanent brain damage. Electrical stimulation causes the activation of nontarget cells in addition to the specific cell types. Thus, scientists could not understand clearly which cells were involved in the circuit. “For the study of neuroscience, optical techniques are more useful tools than the pharmaceutical and the electrical ones because of their higher speed and accuracy and less damage to tissue. It can optionally stimulate or silence particular cell types and neuronal circuits with millisecond temporal (related to time) accuracy. This method allows more temporal resolution for analyzing a specific neural circuit operation in different diseases.” (10) 

Future implications of Optogenetics.

·      Ethical considerations of its memory modifying potential

As discussed before, optogenetic techniques can be used to modify memory.  They can blunt emotional (traumatic) memories but also change their valences from negative to positive and vice versa. In such cases, the issue of exploitation seems to be of particular importance, as political regimes could use optogenetics for their own purposes. “On one hand, this concern appears graver in the case of changing a memory’s valence, as opposed to erasure of a specific memory, as authoritarian governments may wish to use war veterans as propaganda tools by imposing positive valences on extremely negative, traumatic war memories.” Thus, people in positions of power could possibly mould the reality of people.  “On the other hand, the issue of exploitation may prove even more disturbing in cases where optogenetics is used to erase memories of, e.g., brutal interrogations of prisoners of war.”(11) Having control over the memory of people would give unprecedented control to powerful people to get away with whatever they wish.

·      Refining cures of neurological disorders

In the realm of finding cures for diseases, “the ability to optically control neural activity opens up possibilities for the restoration of normal function following neurological disorders. The temporal precision, spatial resolution, and neuronal specificity that optogenetics offers is unequalled by other available methods.” “Optogenetics is not yet suitable for use in humans. Instead we conclude that for the immediate future, optogenetics is the neurological equivalent of the 3D printer: its flexibility providing an ideal tool for testing and prototyping solutions for treating brain disorders and augmenting brain function.” (14) Thus, it could shed light into the mystery of the black box that is our brain.

As Optogenetics is not yet used for humans, the possible treatments of neurodegenerative diseases would provide a more refined approach to solving these problems with little to no side effects. “Optogenetics is quickly becoming one of the most powerful tools in neuroscience. It will not only shed light on functional brain mapping, it will increase our understanding of complex neural diseases potentially leading to revolutionary treatments.” (15)

 

Bravo! You completed the article. I hope you could takeaway some of what we learnt about the novel technology of Optogenetics. Stay curious!

 

 

Bibliography :

Article references :

Wikipedia: Cell(Biology), types of cell

Introduction to the nervous system- SEER Training (3)

Sensation meaning: Collins dictionary

What are thoughts made of? : MIT School of Engineering

What Is Optogenetics and How Can We Use It to Discover More About the Brain? By frontiers for Young minds (1)

 

Metabolism and weight loss: How you burn calories? – Mayo Clinic (2)

Growth Definition and Meaning: Dictionary

Reproduction: Wikipedia

Gene: Genome (4)

What is DNA? MedlinePlus Genetics

What is Algae? : LiveScience

Eyespot Apparatus: Wikipedia

Photosynthesis :National Geographic Society

Structure and types of flagella : Byju’s

Optical Fibre: Wikipedia (5)

Epilepsy: Apollo Hospitals (6)

Spatiotemporal mechanisms of life: Nature (7)

Generation of the circadian rhythms in the suprachiasmatic nucleus: Nature (8)

Where is the suprachiasmatic nucleus? : Neuroscientifically Challenged (9)

Optogenetics, Tools and Applications in Neurobiology

-         NCBI (10)

Optogenetics: Britannica (13)

Current and future applications of Optogenetics – News Medical

The Memory-modifying Potential of Optogenetics and the need for neuroethics : Springer Link (11)

Opto Definition and Meaning: Dictionary

Neurodegenerative diseases  : NIEHS.NIH.GOV

Deep brain stimulation: John Hopkins Medicine

Optogenetics for neurodegenerative diseases: NCBI.NLM.NIH.GOV

Neural grafting: Today, tomorrow or never? : Science Direct (12)

Prospects for Optogenetic Augmentation of Brain Function: frontiers in Systems Neuroscience (14)

Optogenetics and the future of brain mapping: Lions talk science (15)

Neuron action potentials: The creation of a brain signal – Khan Academy (16)

Video references:

What investigating neural pathways can reveal about mental health by Kay M. Tye : TED@NAS

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