A Search for the Dying Neurons: An

Analysis
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The mitogen activated protein kinase

(MAPK) pathway plays a vital role in the
cell cycle, growth, and differentiation.
Germline monogenic (single gene)
mutations in genes that code for proteins in
the pathway can lead to a family of disorders
called RASopathies. The family of
RASopathies includes Noonan Syndrome,
Neurofibromatosis Type 1, Cardio-faciocutaneous Syndrome, and LEOPARD

contribute to neural circuit defects and

possibly GABAergic neuron loss.

Syndrome among others, and they share

many similar phenotypic characteristics such
as dysmorphic facial features, cardiac
defects, and a higher occurrence of cancer.
Most of the mutations in genes that encode
for proteins in the pathway lead to gain-offunction (hyperactivation) in the MAPK
pathway [1]. A previous study has recently
found that expression of a gain- of-function
constitutively active MEK1 (a protein
downstream in the MAPK pathway)
mutation in mice leads to the loss of ~60%
of the cortical parvalbumin (PV) subset of
inhibitory interneurons by P14. We
hypothesize that hyperactivation of the
MAPK pathway disrupts the morphology of
PV interneurons, a response that could

morphological and biochemical pathology is

necessary. Dendritic morphology as well as
somal morphology is a key factor that
regulates the amount of input into neurons.
Therefore determining whether changes in
input occur in response to these mutations
may assist in understanding deficits in
neural circuit function. Defects in
parvalbumin GABAergic neurons have been
implicated in of ASDs, schizophrenia, and
traumatic brain injury, so this research can
extend far beyond the scope of determined
RASopathies. In a subset of these conditions
(ASDs, schizophrenia, traumatic brain
injury) aberrant MAPK signaling has been
observed. Thus, deciphering how MAPK
regulates this critical neuronal cell type may

So why exactly did I do this project? Well

for a few reasons. RASopathies, syndromes
induced by mutations in the MAPK
pathway, have a combined incidence of
about 1:2000-1:3000 births [1]. In order to
understand and possibly reverse the effects
of these syndromes, identification of the

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further our understanding of a wide range of

pathological states [2,3]

Dendrite Length Analysis

After completing a fairly extensive

procedure, I was able to do an analysis on
the resulting images and scans. Below is an
image of a vibrotome, the machine that I
used in order to section my mouse brain
tissue. I then used a 6-well tray to do my
staining procedure. After conducting my
experiment, I was able to draw some
conclusions. They are outlined next. It was
previously determined that CA-MEK1
caused a 50% decrease in parvalbumin (PV)
neurons in P14 mice, and this study
furthered the previous one by determining
that not only did this 50% decrease in PV
neurons occur, but there appeared to be an

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alteration in the dendritic morphology of

these surviving PV interneurons. Although
the somal sizes of the mutants did not seem
altered compared to the controls, there was a
definite increase in the dendritic
morphology, both in the dendrite length and
the number of dendritic nodes. These
measures combined offer a measure of
dendritic complexity.
PV interneurons, which are shaped
like basket-cells, are known to be fastspiking neurons in the brain. Previous
research has revealed that size of a neuron
often can clue into the function of these
neurons. Therefore, we see that hyperactive
MAPK function may cause increased
dendritic branching in parvalbumin-

expressing inhibitory interneurons.

In terms of future research, further
characterization of these parvalbumin
interneurons extremely necessary. In
addition, electrophysiological recordings
should be utilized in the future, as they
would give a more direct and accurate value
that assesses the amount of synaptic input
that cells are receiving. Also, on a larger
scope, after identification of the effects of
hyperactivation of MAPK on inhibitory
interneurons is complete, possible
therapeutic approaches towards these effects
should be developed to reverse the effects of
some of these syndromes.

References
A

1. Tidyman, William E., and Katherine A. Rauen. "The RASopathies: Developmental

Syndromes of Ras/MAPK Pathway Dysregulation."Current Opinion in Genetics &
Development 19.3 (2009): 230-36. Web.

2. Snow, Wanda M., Kelly Hartle, and Tammy L. Ivanco. "Altered Morphology of Motor
Cortex Neurons in the VPA Rat Model of Autism."Developmental Psychobiology 50.7
(2008): 633-39. Web

3. Mndez, Pablo, and Alberto Bacci. "Assortment of GABAergic Plasticity in the

Cortical Interneuron Melting Pot." Neural Plasticity 2011 (2011): 1-14. Web.

4. Lawrence, Y. A., T. L. Kemper, M. L. Bauman, and G. J. Blatt. "Parvalbumin-,

Calbindin-, and Calretinin-immunoreactive Hippocampal Interneuron Density in
Autism." Acta Neurologica Scandinavica 121.2 (2010): 99-108. Web.

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