Connie Chen

IR2-10-GT
April 30, 2019
Data collection
Turn in #5
Reconnecting the Brain and the Body Literature Review: Axon Regeneration in Spinal Cord
Injuries.

Part 1:
1) Topic: Axon Regeneration after spinal cord injury is believed to be critical for full
functional recovery, but normally occurs extremely rarely in nature. Scientists have
worked tirelessly throughout the years to discover which factors assist in the regeneration
and which factors inhibit. This literature review explores and analyzes the different
studies conducted that resulted in the discovery of the importance of many factors.
2) Method: A literature review was conducted to aid in the collection of the data. This was
the best method to choose for the topic because the depth of the biology requires
expertise and a simple questionnaire for the basic public would not yield results.
Additionally, a literature review would offer a more complex analysis and help provide a
deeper understanding of the topic.
3) Distribution plan: The plan is to incorporate the analysis from the literature review into
my paper to offer a deeper dive in the the biologically heavy topic. My paper will then be
available on my online portfolio if someone did want to access it. However, I do not plan
on publishing my paper because it would not contribute anything new into the field.
Part 2: Citations for studies
1) Richardson, P. M., McGuinness, U. M., & Aguayo, A. J. (1980). Axons from CNS
neurones regenerate into PNS grafts. ​Nature,​ ​284(​ 5753), 264.
https://doi.org/10.1038/284264a0
2) Geoffroy, C. G., Lorenzana, A. O., Kwan, J. P., Lin, K., Ghassemi, O., Ma, A., … Zheng,
B. (2015). Effects of PTEN and Nogo Codeletion on Corticospinal Axon Sprouting and
Regeneration in Mice. ​The Journal of Neuroscience​, ​35​(16), 6413–6428.
https://doi.org/10.1523/JNEUROSCI.4013-14.2015
3) Cheah, M., Andrews, M. R., Chew, D. J., Moloney, E. B., Verhaagen, J., Fässler, R., &
Fawcett, J. W. (2016). Expression of an Activated Integrin Promotes Long-Distance
Sensory Axon Regeneration in the Spinal Cord. ​The Journal of Neuroscience​, ​36​(27),
7283–7297. ​https://doi.org/10.1523/JNEUROSCI.0901-16.2016
 Study Author(s) Experimental Conclusions
and Date of Procedure
Publication

Axons from CNS neurones regenerate into PNS grafts P. M. “In the first group of “The
Richardson, 16 animals, a 5-mm perineurium surrounding the common
U. M. segment of the peroneal and posterior
McGuinness midthoracic spinal cord tibial fascicles of the sciatic nerve was
& A. J. was removed and an clearly identifiable in
Aguayo autologous sciatic cross-sections through the middle of the
nerve graft was graft and contained
March 20, inserted sub-pially many myelinated or unmyelinated axons
1980 between the two ensheathed by
stumps of Schwann cells. The ultrastructural
the spinal cord. One to appearance of each graft was
four months later, the similar to that of a regenerated peripheral
animals were nerve. The grafts were
anaesthetised and in gross continuity with the proximal and
perfused. The grafts distal spinal cord
and adjacent spinal stumps.”
cord
tissue were examined
by light and electron
microscopy.”

Effects of PTEN and Nogo Codeletion on Corticospinal Geoffrey G. Mice Sample​: “The primary goal of this study was to
Axon Sprouting and Regeneration in Mice Cédric, “AAV-Cre or control assess whether manipulating both
Lorenzana virus AAV-GFP were neuronintrinsic (PTEN) and extrinsic
O. Ariana, injected into the (Nogo) factors could further increase the
Kwan P. sensorimotor cortex of sprouting and regeneration of CST axons
Jeffrey, Lin P1 or young adult (4- following injury beyond what is already
Kyle, to 6-week-old) mice to observed with manipulating either factor
Ghassemi obtain the following alone.”
Omeed, Ma four experimental “Nogo deletion did not further increase
Andrew, Xu groups: control mice CST sprouting that is already enhanced by
Nuo, Creger (PTENf/f;Nogo/ , neonatal PTEN deletion”
Daniel, Liu AAV-GFP), Nogo “Nogo deletion did not further enhance
Kai, He single-mutant mice CST sprouting that is already enhanced by
Zhigang, (PTENf/f;Nogo /, young adult PTEN deletion”
Zheng AAV-GFP), PTEN “Nogo deletion further enhanced CST
Binhai singlemutant mice regeneration elicited by neonatal PTEN
(PTEN /;Nogo/ , deletion”
April 22, AAV-Cre), and “Nogo-A is upregulated at and around the
2015 PTEN/Nogo injury site”
doublemutant mice “Nogo deletion further enhanced CST
(PTEN /;Nogo /, regeneration elicited by young adult PTEN
AAV-Cre).” deletion”
Tests Run: ​1)“This
test assesses the
forepaw preference for
touching and leaning
against the wall when
mice explore around in
a new cage while
standing on their
hindlimbs”2)“Mice
were habituated to the
testing environment by
being handled as they
would during testing to
 decrease the risk of
anxiety. This consisted
of handling the mice
and touching their
forepaws as it would be
done to attach a sticker
for the actual test. Mice
were then left for 2 min
in a
cylinder.”3)“Briefly,
mice were habituated
and trained to run on
the apparatus for 5
consecutive days
before injury. On the
day before injury and
on selected days after
injury, mice were
tested. An irregular
rung pattern (with
uneven spacing
between rungs) is used
each time to prevent
the mice from
compensating for their
limb impairment by
learning a particular
pattern.”4)“Mice were
observed for 5 min by
two observers. Many
features were noted,
including ankle
movements, stepping
pattern, coordination,
paw placement, trunk
instability, and tail
position, with a
minimum score of 0
(no movement) to a
maximum score of 9
(normal
locomotion).”5) “Mice
were placed on a rod
(Ugo Basile) rotating at
increasing speeds from
5 to 50 rpm in 3 min
with constant
acceleration as
described previously
(Lee et al., 2010). The
latency to fall (in
seconds) was averaged
between two trials per
session”

Expression of an Activated Integrin Promotes Cheah, Experimental Group: “Expression of both 9 integrin and
Long-Distance Sensory Axon Regeneration in the Spinal Menghon, “Adult female Sprague kindlin-1 promotes DRG neurite outgrowth
Cord Andrews, Dawley rats were on tenascin-C and aggrecan”
Melissa R., killed, and DRGs were “Expression of both 9 integrin and
Chew, harvested. For explant kindlin-1 promotes sensory axon
 Daniel J., culture, each DRG was regeneration in vivo in a timedependent
Moloney, cut into two to three manner”
Elizabeth B., pieces and then plated “To assess ingrowth into the cord from the
Verhaagen, on substrate-coated virus-injected C7 DRG, we analyzed axon
Joost, glass coverslips. For regeneration in the dorsal root, dorsal root
Fässler, dissociated culture, entry zone (DREZ), and C7 spinal
Reinhard, DRGs were incubated cord...Within the dorsal root, all groups
Fawcett, with 0.2% collagenase had 47 regenerated axons per 14 m section
James W. (Sigma) and 0.1% (three sections per animal, n 10) distal to
trypsin (Sigma)” the crush site with no significant difference
July 6, 2016 Tests Run:​ 1) found between groups...Within the DREZ,
“Ladder-rung walking all three experimental groups, 9-only (12 3
task, mechanical axons), kindlin1-only (15 4 axons), and 9
pressure kindlin1 (13 3 axons), showed
(Randall–Selitto touch significantly more axons per 14 m section
sensitivity) test, and (three sections per animal, *p 0.0466,
thermal pain one-way ANOVA, n 10) than the fGFP
(Hargreave’s hotplate) control group... In the C7 spinal cord
test were administered segment, both the kindlin1-only (22 5
once before the surgery axons) and 9 kindlin1 (48 10 axons)
and once each week groups had significantly more axons per
after surgery for 12 section (three sections per animal, ***p
weeks.”2) “Animals 7.99e 4 , one-way ANOVA, n 10) than the
were placed in an fGFP control (0.3 0.2 axons) and 9-only (2
enclosure with a metal 0.5 axons) groups... Furthermore, the 9
mesh bottom. A probe kindlin1 group had significantly more
connected to an axons per section (*p 0.0361, one-way
electronic ANOVA, n 10) than the kindlin1-only
anesthesiometer (IITC group. Axons expressing both 9 integrin
Life Science) was and kindlin-1 were observed in the spinal
applied with a gradual cord, especially in the cuneate fasciculus
increase of pressure to and dorsal horn...For the fGFP control
the footpad of the group, no axons were observed growing
forepaw, until the into the DREZ and spinal cord”
animal withdrew its
paw.”3) “Animals were
placed in an enclosure
with a fiberglass
bottom. An
infraredemitting device
(Ugo Basile) was
placed directly under
the footpad of the
forepaw, until the
animal withdrew its
paw”

Part 3: Analysis and Results

1) All three factors, the peripheral nervous system environment, Nogo and PTEN, and 9
integrin and Kindlin-1, greatly affect axon regeneration either in beneficial or harmful
ways. In the Aguayo study, the researchers came to the conclusion that environmental
factors in the Peripheral nervous system are much more beneficial to the regeneration of
axons after injury. In the study regarding Nogo and PTEN, the researchers were able to
 draw conclusions about Nogo’s effect on axon regeneration in regard to PTEN deletion.
And in the study regarding integrin, the researchers were able to conclude the how 9
integrin and kindlin-1 affect axon regeneration in different settings. All three of theses
studies showcase various factors that impact axon regeneration, how they impact it, and
in what circumstances they do. Additionally, all three studies were conducted on rats due
to their similarities to humans on a molecular level.
2) However, these studies are difficult to compare due to the variety of factors that were
experimented on and also because of the different effects that were studied. The three
studies separately do not offer any definite way for axons to regenerate after injury, put
together in a combined treatment may prove to be helpful. This is why it is important to
study all aspects of axon regeneration and not focus on one, because only changing one
factor will not result in full functional recovery. It is not known at this point if full
functional recovery is even possible, but the way to be able to achieve close to it is
through the manipulation of several different factors that affect axon regeneration.