Andrew Qian

IR-3/10GT
1/30/18
#12
TBI: Neurogenesis and the Key to Recovery

Traumatic Brain Injury (TBI) affects approximately 2.8 million people in the United

States each year, and nearly 50,000 of those people will die from TBI related causes (“TBI: Get

the Facts” 4). TBI causes 30% of all injury-related deaths, culminating to approximately 153

deaths per day. Even those who sustain non-fatal brain injuries experience chronic conditions for

the rest of their lives. Those who survive are at risk of suffering from life long disabilities, such

as impaired thinking or memory, loss of motor control, and changes in personality “Facts About

Traumatic Brain Injury” 16).These chronic issues not only affect the individuals with TBI, but

also have lasting effects on their families and communities. Today, an estimated 5.3 million men,

women, and children are living with a permanent TBI-related disability in the United States

(“How Many People Are Affected” 12). Traumatic brain injuries pose a serious problem to

people’s well-being. Currently, no efficient method to treat traumatic brain injuries exists;

however, current research on astrocytes shows potential in treating TBI. Evidence points to the

possibility that stimulating astrocytes will allow for increased cell communications among brain

cells, accelerated neurogenesis, and increased efficiency of recovery of the brain after TBI.

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 TBI is a term used to describe what occurs when the brain is injured from an external

force that disrupts normal functions. The damage ranges from mild trauma, which is simply a

brief change in brain conscious, to severe trauma, which involves an extensive period of

unconsciousness, resulting in possible memory loss. After TBI, the visible damage is

accompanied by deterioration on the cellular level. In the brain, there are extensive connections

between neurons and glial cells, allowing more communication between brain cells in order to

control the many bodily functions through neurotransmitters. TBI causes the death of these

neurons and glial cells, as well as the cellular extensions such as dendrites and axons (Pekna 18).

At the time of injury, mechanical perturbation of neurons is associated with a significant

release of a host of neurotransmitters particularly the release of glutamate and other excitatory

amino acids. This release of neurotransmitters results in an influx of extracellular Ca2+ into the

cell; in turn, additional Ca2+ is released from intracellular stores, thus producing sufficient

quantities of free intracellular Ca2+. This intracellular Ca2+ then initiates a host of intracellular

reactions, which can result in cytotoxic injury and eventually cell death (McAllister 9). This

disrupts the connections and prevents the signaling that regulates important functions of the

body.

Astrocytes are a type of glial cell, which comprise nearly 90% of brain cells. Scientists

originally thought that glial cell were merely supporting cells, but recent findings show that they

play a much bigger role in the brain than scientists originally believed. First, the sheer number of

astrocytes and the structure of these cells in the brain allow them to communicate with many

other cells in the brain. As described by Dr. Andrew Koob, astrocytes have “hundreds of

'endfeet' spreading out from their body,” enabling them to send calcium signals down these

extensions to surrounding (“The Root of Thought” 7). This particular structure allows them to

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play a role in numerous homeostatic processes. The shape of the astrocytes, as previously

described as containing “hundreds of endfeet”, allows them to support the structure of the brain

parenchyma. Additionally, astrocytes also release nutritional and energy substrates into the brain;

this characteristic highlights the key role that they play in supporting other brain cells

(“Astrocytes: What are they” 6).

Astrocytes can aid in neuronal development as well, forming molecular boundaries that

take part in guiding the migration of developing axons and certain neuroblasts. In addition,

recent research indicates that astrocytes are essential for the formation of synapses in the brain.

In studies done by Dr. Christopherson and Dr. Barres, they identified certain chemical signalings

that were indispensible for the development of synapses, such as thrombospondin. They also

found that astrocytes also release signals that tag certain synapses for elimination, thus allowing

for “synaptic pruning”. Astrocytes are also involved in other neuronal mechanisms such as flow

of blood through the brain. The structure of an astrocyte has extensive contact with blood vessels

that allows them to have multiple interactions with other brain cells. Studies have shown that

astrocytes are able to produce numerous distinct mediators such as prostaglandins and nitric

oxide that can widen CNS blood vessels, as well as modulate blood flow direction (Sonfroniew,

Vinters 18). The discovery of these chemical signals all lead to the conclusion that astrocytes

play an essential role in the brain; this information could be used to develop an effective

treatment method for TBI.

Neurogenesis refers to the growth and development of nervous tissue. This process is

most active while a baby is in the womb, and is responsible for the creation of neurons. Scientists

previously believed that neurogenesis terminates during adulthood; however, in the 20th century,

further research revealed that neurogenesis does occur in some areas of the adult brain, including

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the subventricular zone (SVZ) and the dentate gyrus of the hippocampus area. In a study

conducted by Dr. Cameron and Dr. Mcvay, the doctors determined that, in the rat dentate gyrus,

approximately “9,000 new cells [are created] per day, which equates to ~270,000 cells per

month.” The continuous formation of new neurons allows for both constant maintenance and

efficient rehabitulation of damaged areas. (Mandal 2).

Adult neurogenesis has the potential as a method for repairing the brain after TBI.

Studies have shown that the adult hippocampus is involved in learning and memory, and

diminished hippocampus neurogenesis is associated with poor, long-term, spatial memory

function. Therefore, neurogenesis contributes to numerous vital bodily functions and allows the

brain to repair itself after injury. Various techniques have proven to increase neurogenesis,

improving function and accelerating post-TBI brain recovery. For example, Dr. Dong Sun

conducted a study in which he identified that direct administration of the growth factors bFGF or

EGF may significantly enhance TBI-induced proliferation in the SVZ and hippocampus,

allowing for drastic improvement of cognitive functional recovery. Other studies, like the one

conducted by Dr. Lu and associates, determined that the enhancement of neurogenesis through

clinical drugs leads to increased cognitive function in statins after TBI (Sun 17). With a newly-

developed method to effectively induce neurogenesis in the brain, patients may be able to

recover from TBI effectively and efficiently.

Stimulation of Astrocytes:

The development of calcium imaging and advanced optical microscopy techniques has

allowed for increased understanding of astrocyte functioning. Through calcium imaging, the

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discovery was made that astrocytes exhibit varying levels of intracellular calcium concentration,

which can spread to surrounding astrocytes. Advanced microscopes have also contributed to the

improvement of analysis in these areas. Two-photon microscopy has enabled scientists to

visualize molecular behaviors beneath the plasma membrane, resulting in the clarification of

calcium interactions in the brain. Collected evidence suggests that that triphosphate mediated

calcium release from the endoplasmic reticulum causes [Ca2+]i increases in astrocytes in

response to activity from nearby neurons and astrocytes, which in turn induces the release of

gliotransmitters, chemical signals that bind to their respective receptors on neurons and modulate

firing frequencies, synaptic transmissions, etc. These gliotransmitters play crucial roles in

ensuring proper brain function. In a study conducted by Dr. Kazuki Harada, he showed that

dysfunctional gliotransmitters can result in serious brain disorders and abnormal behavior,

leading to the conclusion that these gliotransmitters are essential for proper brain function.

(Harada, Kamiya, Tsuboi 12).

There are various hypothesized methods in which gliotransmitters are thought to be

released. It is thought that they can be released from a storage compartment through exocytosis

or directly from the cytosol through plasma membrane ion channels. As these two methods have

different benefits and drawbacks, it is hypothesized that they may be used for different purposes.

Identification of the different methods of gliotransmitter release is important, as it would allow

scientist to develop a better treatment method for TBI.

Release through ion channels is thought to be more energy efficient in comparison to

release through a vesicle against its concentration gradient, but releases the gliotransmitter in

much smaller quantities compared to exocytosis. In the case of exocytosis, the SNARE Protein

family is essential for the regulation of the vesicles that are responsible for transporting

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gliotransmitters out of the cell. Vesicle associated membrane proteins, otherwise known as

VAMPs, are a family of SNARE protein that are mostly involved in vesicle fusion Multiple

studies using mice models have been used to analyze whether or not exocytosis underlies the

modulatory actions of astrocytes. Studies have been done that use tetanus or botulinum toxins

that cleave VAMP2/3 or NAP 23/25 selectively to determine whether or not exocytosis is

important for gliotransmitters. The results of said experiments show that the synaptic effects of

gliotransmission were blocked, indicating that exocytosis may be one mechanism in which

gliotransmitters are released from astrocytes. Studies have also been done based on the

possibility of protein channels as a method for gliotransmitter release. Anion channels have been

the main focus in this area, as they have been shown to allow flux of both small, inorganic

anions, such as chlorine, as well as larger molecules such as taurine, and amino acids. Studies

have determined that the anion channel Best-1 functions in GABA release from cerebellar glia,

as well as glutamate release in the CA1 hippocampus, two gliotransmitters that are vital for

modulating processes in the brain (Sahlender, Savtchouk, Volterra 9). By determining what the

most effective method of astrocyte stimulation is and how gliotransmitters are released from the

astrocyte, more effective treatment methods for TBI can be produced.

Neuronal Signaling:

The role that signaling plays within the brain is immense as it induces many different

functions of the brain and body. Today, it is widely accepted that glial cells exhibit G-Protein

Coupled Receptors (GCPR) that are similarly expressed in neurons, and that the stimulation of

these receptors evokes a variety of responses, the primary being intracellular calcium

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concentration. Once the GCPR’s are activated, it leads to the activation of phospholipase C, a

class of enzymes that cleave phosphates. This then results in the production of IP3, which leads

to Ca2+ release from the Endoplasmic Reticulum (Scemes, Giaume 10). Astrocytes in particular

exhibit a large number of GPCRs linked to calcium mobilization from internal stores. Since this

discovery, multiple laboratories have reported that calcium elevations may result in the release of

gliotransmitters, including glutamate, ATP, and D-serine, all of which bind to “postsynaptic

neuronal receptors to modulate synaptic transmission and other activity” (Agulhon et al. 7).

Calcium signaling plays a major role in neuronal signaling throughout the brain, and its ability to

stimulate gliotransmitter release from astrocytes makes it a valuable area of research in

developing a competent treatment for TBI.

Another major form of signaling that plays an important role throughout the brain is

ephrin signaling. Ephrins are a family of proteins that bind to eph receptors, which are tyrosine

kinase receptors. Studies on this type of receptor have revealed that they have numerous roles in

the brain, both in regulation and activation of neuronal processes. It has been discovered that eph

signaling can also stimulate gliotransmitter release in astrocytes, indicating the broader role that

they may play in regulating synaptic processes. In the brain, EphrinB3 and ephrinB1 stimulate

astrocytic release of D-serine through the dephosphorylation of PKCα and activation of serine

racemase; this then converts L-serine to D-serine. This stimulation is due to the ephrinB3 and

ephrinB1 interactions with the EphB3 and EphA4 receptors. Experimentation showed that in the

absence of EphB3 and EphA4, there was reduced D-serine release by the astrocyte. Using high-

performance liquid chromatography (HPLC) to measure glutamine levels, it was also found that

ephrinB3, but not ephrinB1, can stimulate glutamine release from astrocytes (Zhuang et al. 12),

(Sahlender, Savtchouk, Volterra 13). As both glutamine and D-serine play essential roles in

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synaptic transmission and other neuronal processes, these results indicate that ephrin signaling is

vitally important in to the proper functioning of the brain.

Ephrin signaling also has a role in neuronal development as well. Studies have shown

that Ephrin - B2 presented by hippocampal astrocytes actually regulates neurogenesis in the adult

hippocampus. Experiments have already determined that ephrins play a role in neuronal stem cell

development at early ages, when the brain is still developing. For example, it has been found

that ephrin-A/EphA signaling promotes embryonic telencephalic NSC differentiation, while

ephrin-B3/EphB3 signaling may inhibit NSC proliferation in the developing subventricular zone

of the brain. Recent studies have shown that Ephrin signaling expressed by astrocytes can

regulate neurogenesis in the adult hippocampus as well. An experiment ran discovered that

ephrin-B2 presented from hippocampal astrocytes regulates neurogenesis in the body, allowing

researchers to determine that Ephrin-B2 promote neuronal differentiation of adult neural stem

cells through juxtacrine signaling (Ashton et al. 18). Evidently, intracellular signaling through

ephrin signaling is crucial for numerous neuronal processes. Better understanding of the different

chemical signals that modulate TBI allows for development of an effective treatment for TBI.

There are other forms of signaling expressed by astrocytes that regulate neurogenesis in

the brain as well. Studies have shown that astrocytes can negatively regulate neurogenesis as

well, which is where neurogenesis is inhibited through cell to cell signaling. Studies have found

that astrocytes can negatively regulate neurogenesis through the the Notch pathway, a cell

signaling system that is important for determining cell fate. In those studies, researchers found

that in both cultures and mice, the absence of glial fibrillary acidic protein (GFAP) and vimentin

(GFAP−/−Vim−/−). They also found that when there was reduced endocytosis of Notch LIgand

Jagged1, there was less notch signaling in the GFAP−/−Vim−/− astrocyte and, therefore, more

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neuronal differentiation. Thus, it can be concluded that astrocytes negatively regulate

neurogenesis through the notch pathway, and the endocytosis of the Jagged1 Notch receptor

ligand is modulated by GFAP and vimentin (Wilhelmsson et al. 12). Intracellular signaling

within the brain is an essential aspect of how neurogenesis is induced and regulated, and a

comprehensive understanding of this communication will be vital for the development of an

effective treatment for TBI.

Finally, there is the ability of astrocytes to modulate NSC neuronal differentiation

through certain genes. In a study done by Dr. Barkho, he found that certain membrane associated

and secreted factors released by astrocytes would greatly promote neuronal differentiation, while

other factors would actually inhibit said differentiation. Dr. Barko found that primarily, it was

Newborn Hippocampal (NBH), adult hippocampal (ADH), and newborn spinal cord (NBS)

astrocytes that released differentiation promoting factors. In contrast, he also found that

astrocytes isolated from non-neurogenic adult spinal cord (ADS) astrocytes and adult skin

fibroblasts (ASF) that exhibited inhibitory effects on NSPC neuronal differentiation. By

comparing the two types, Dr. Barko confirmed that both “IGFBP6 and decorin, inhibitors of IGF

and TGFβ2 respectively ... inhibited neuronal differentiation of NSPCs that were cultured with

NBH astrocytes.” Surprisingly, he also discovered that at low concentrations, IL-1β and IL-6

could promote NSC differentiation as well. This was unexpected as previously, it was commonly

believed that these two cytokines only inhibited differentiation (Barkho 6). By identifying the

genes that are responsible for this distinction in astrocytic modulation, researchers can gain a

better understanding of the molecular aspects that regulate adult neurogenesis, allowing them to

develop a better treatment method for TBI.

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 Evidently, TBI is a major issue in the United States. It is important for researchers to

develop a treatment method as early as possible as it will allow people to recover from chronic

injuries. Currently, astrocytes contain an immense potential to be a valuable area of research.

Their location in the brain, and its control over various neuronal pathways and receptors allow it

be a valuable factor of many different neuronal processes. The structure of the astrocyte is very

unique, with many different endfeet, so it is able to be in contact with many different cell in the

brain. This allows them not only to play a supporting role in regulating things like providing

vitamins and blood flow, but they can also send chemical signals throughout the cell promoting

development of the brain en vivo. With further research, it is not unreasonable to suppose that

stimulation of astrocytes will allow for increased cell communications among brain cells,

accelerated neurogenesis, and therefore increased efficiency of recovery of the brain after TBI.

In view of these findings, Astrocytes could be the key in discovering an efficient method to

induce neurogenesis in the brain, creating an efficient method for treating TBI.

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Vocabulary:

● G Protein Receptor: A type of receptor that is attached to G proteins;

Controls many functions of the body such as hearing, sight, etc.

● Ephrin Receptors: A kind of receptor that generally modulates neuronal

processes in the brain

● Neurotransmitter: chemical messengers that enable neurotransmission.

● NSC: Neuronal Stem Cell, a type of cell in the brain that can differentiate

into another kind of cell.

● En vivo: In the body

● Notch1:a single-pass transmembrane receptor.

● Neurogenesis: the growth and development of nervous tissue.

● Glutamate: A type of chemical siangla that asks as both a neurotransmitter

and a gliotransmitter; responsible for sending numerous signals throughout

the brain.

● Exocytosis: Release of an object from a cell through vesicle transport.

● Cytokines: A broad category of proteins whose release has an effect on the

cells around them.

● TBI: A non degenerative, non congenital insult to the brain from an

external mechanical force, possibly leading to permanent or temporary
impairment of cognitive, physical, and psychosocial functions, with an
associated diminished or altered state of consciousness. (When the brain is
injured from an external force.)

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