Review Article

The Potential of Astrocyte Modulated

Ephrin Signaling for Enhancement of
Neurogenesis
Andrew Qian
River Hill High School

Abstract:
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).
In fact, 30% of all injury-related deaths are a result of TBI, which is approximately 153 deaths per
day. Even those who sustain non-fatal brain injuries experience chronic conditions for the rest of
their lives, 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). Clearly, TBI poses a severe problem to people’s
well-being. As of now, no efficient method to treat traumatic brain injuries exists; however, current
research on astrocytes shows potential in treating TBI. Furthermore, the role of neurogenesis has
also been explored in developing medicinal treatments for the damage that results from TBI.
Current evidence points to the possibility that stimulating astrocytes will allow for increased
regulation of ephrin signaling, possible leading to accelerated neurogenesis and therefore more
effective recovery of the brain after TBI.
Introduction:

The vulnerability of the brain poses a problem to long-term neuronal sustainability as

various external insults can sever connections throughout the brain, thus impairing cognitive ability
or motor control. This condition is known as Traumatic Brain Injury. To effectively recover from
such injury, it requires the growth and formation of new neuronal cells, as well as the development
of new connections between new neurons. One proposed solution for repairing the brain after TBI
is through increasing neuronal differentiation. This idea involves replenishing damaged neurons
through proliferation of adult NSPC’s residing in the brain. Recent evidence suggests that glial
cells, specifically astrocytes, could potentially play a major role in modulating such neuronal
differentiation.

The neuronal damage from TBI ranges from mild trauma, which is simply a brief change in
brain consciousness, 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 many bodily functions
through neurotransmitters (Pekna). TBI results in the severing of such connections. At the time of
injury, neuronal damage can lead to mechanical perturbation of neurons, which in turn is
associated with a significant release of neurotransmitters, particularly glutamate and other
excitatory amino acids. This release of neurotransmitters causes extracellular Ca to rush into the
2+

cell; in turn, additional Ca is released from stores within the cell, thus producing a considerable
2+

quantity of intracellular Ca . This Ca then initiates a set of intracellular reactions, which can result
2+ 2+

in cytotoxic injury and eventually cell death (McAllister). Traditionally, TBI is treated through
therapeutic methods, in addition to surgical treatment to reduce swelling and pressure within the
brain at the spot of injury. Currently, there are no treatment methods which directly focus on
astrocytes in repairing the brain after cellular injury from TBI.

What are Astrocytes and Ephrin proteins?

Astrocytes are a type of glial cell which comprise nearly 90% of brain cells. Scientists
originally thought that glial cells were merely supporting cells, but recent findings reveal that they
play a much bigger role in the brain than originally believed (“Astrocytes: What are they”). 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 through juxtracrine signaling by cellular contact. 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 cells (“The Root of
Thought”). This structure allows them to play a role in numerous homeostatic processes, such as
support of the structure of the brain parenchyma. Additionally, recent research indicates that
astrocytes are essential for the formation of synapses in the brain. Studies have identified astrocytic
release of certain chemical signal that are indispensable 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”. Finally, astrocytes are also involved in other
neuronal mechanisms such as neuronal blood flow. The aforementioned “end feet” of an astrocyte
has extensive contact with neuronal blood vessels that allows them to have multiple regulatory
interactions with blood vessels. 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

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well as modulate blood flow direction (Sonfroniew, Vinters 18). The prominence of astrocyte
involvement in neuronal development and maintenance suggest the possibility of astrocytes as
targets in the development of a treatment for TBI.

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, both EphrinB3 and ephrinB1 can stimulate
astrocytic release of D-serine, a gliotransmitter, through the dephosphorylation of PKCα, as well as
the activation of serine racemase, an enzyme that converts L-serine to D-serine. This stimulation is
due to the ephrinB3 and ephrinB1 interactions with the EphB3 and EphA4 receptors. In the
absence of EphB3 and EphA4, experimentation revealed that the astrocytes released reduced D-
serine levels. In addition, using high-performance liquid chromatography (HPLC) to measure
glutamine levels it found that ephrinB3, but not ephrinB1, can stimulate glutamine release from
astrocytes, showing that ephrin signaling is critical for regulation of gliotransmitter release.
(Zhuang et al.), (Sahlender, Savtchouk, Volterra). As both glutamine and D-serine, as well as
various other gliotransmitters, play essential roles in synaptic transmission and other neuronal
processes, these results indicate that ephrin signaling is vitally important in to the proper
functioning of the brain.

This review focuses on astrocytic modulation of ephrin signaling, and the potential of
ephrins presented by astrocytes in promoting neuronal differentiation. It analyzes recent studies
that have been conducted and examines how astrocytic modulation of ephrin signaling could
rectify TBI through increased neuronal differentiation.

Analysis:

Firstly, the potential of neurogenesis as a method to heal the brain after injury has been
established through multiple studies. Recent experimentation has concluded that neurogenesis is a
constant development throughout the brain, even after maturity. Previous studies have found that
the dentate gyrus of a rate produces around 9000 new cells per day, or approximately 270000 new
cells per month. Similarly, studies have found that in culture, groups of cells that are detached from
the adult human brain can develop new neurons and glia cells, indicating that each cell has the
capability to generate new brain cells. This information has applied in recent studies, where injury
was simulated in mice through various TBI models. These experiments revealed that after injury,
cell proliferation was increased within the sub granular zone and dentate gyrus of the affected
mice. In one such study, which utilized bromodeoxyuridine (BrdU) staining to measure neuronal
proliferation, simulated injury led to the development of 3.5 times as many BrdU cells in
comparison with the sham. Furthermore, this increase in the formation of new cells was
accompanied by integration of newly generated cells into the existing neural circuitry, which
demonstrates that the brain has the capability to rebuild neural connections and reestablish the
neuronal network. This then enables the newly proliferated cells to function as working neurons
and improve cognitive function after injury. This was proven by the previously mentioned study,
where testing after injury revealed that cells with the highest levels of cell growth had the greatest
improvement of cognitive function, which was shown by the shorter latency response of those
mice in the Morris water maze (Sun). These results cumulatively indicate that nerve cells have

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inherent capabilities of recovery and rehabilitation, which could be harnessed in the development
of a more effective treatment for TBI.

Secondly, experimentation has also shown that ephrin signaling plays a vital role in
engendering increased cell proliferation. 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 NSC differentiation within the
embryonic telencephalon, while ephrin-B3/EphB3 signaling may inhibit NSC proliferation in the
developing subventricular zone of the brain. This same trend continues later in life, where various
forms of ephrin signaling can regulate the proliferation, or lack of, neural progenitor cells. While a
normal EphB1 brain showed no visible signs of neuron loss, a brain with EphB1; EphB2 double
mutants showed a noticeable decline in dentate gyrus volume, indicating that EphB1 plays a
critical role in the development of this structure, and to an extent, the rest of the brain. Other forms
of ephrin signaling have also served to regulate neural proliferation. Multiple studies have shown
that Ephrin A; ephA signaling plays a generally inhibitory role in modulating neuronal
proliferation. One such example is ephrin A2, where studies have shown that EphA7>ephrinA2
reverse signaling is able to inhibit SVZ neurogenesis by controlling the cell cycle length of type C
progenitor cells. (Laussu, 12). This information can be used to develop therapeutic treatments as
regulation of ephrin signaling would allow for regulation of neurogenesis in patients, leading to
more effective recoveries.

Finally, astrocytes are potentially able to modulate neurogenesis through ephrin signaling.
Studies have shown that ephrin-B2 released by astrocytes in the hippocampus can induce neuronal
differentiation within the sub granular zone (SGZ) of the adult hippocampus. This is supported by
the results of various experiments. In one study, staining of the brain revealed that hippocampal
astrocytes express ephrin-B2, which is a major ligand in ephrin signaling. Additionally, staining
also revealed that various forms of progenitor cells in the SGZ express EphB4, the receptor for
ephrin B2. This indicates that astrocytes play an important role in regulating neuronal proliferation
of progenitor cells in the brain through ephrin signaling. Secondly, the study also showed that
injection of Fc-ephrin-B2 led to higher levels of cell proliferation, showing that ephrin signaling
plays an important role in modulation of neurogenesis. In this study, Injection of ephrin-B2 ligands
lead to an increase of 55000 BrdU cells in the brain, which was a much larger increase than other
+

vehicle and constituent controls, which only lead increases of less than 30000 BrdU cells in the
+

brain. By connecting this with the results from previous studies, the multiple analysis’ can
conclude that ephrins are a key factor in increasing neural development and developing an
effective treatment for TBI. Additionally, by knowing that ephrin is important for modulating
neuronal differentiation from the information in previous studies, one can conclude that astrocytic
regulation of this signaling pathway through release of ligands is vital for controlling the
differentiation of NSC’s. Furthermore, this signaling has been shown to activate β-catenin, which
in turn induces transcription of proneural transcription factors, inducing neuronal differentiation
(Ashton, 15). Thus, by connecting this information with the previously drawn conclusions, it can
be inferred that astrocytes play a major role in regulating neurogenesis and neuronal repair.
Therefore, controlling astrocytic release of ligands would allow for the stimulation of this neuronal
pathway, and in turn

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Conclusions:

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 make it a
valuable component of many different neuronal processes. The evidence in this review article
suggests that ephrin signaling pathways modulated by astrocytic release of ligands plays a major
role in neurogenesis. Therefore, it is not unreasonable to suppose that stimulation of astrocytes will
allow for adjustment in the ephrin signaling pathway, possibly accelerating neurogenesis
throughout an injured brain, allowing for replacement of damaged neurons, leading to better
recovery for the patient. Further research is required to confirm this theory as well as determine a
method of astrocytic stimulation which would induce accelerated release of ligands. In view of
these findings, however, it can be hypothesized that astrocytes are key in discovering an efficient
method to induce neurogenesis in the brain, creating an efficient method for treating TBI.

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