Undergraduate

Research
Opportunities
Program
2012 Name: Colin Wilhelm
Application Current Street / City / State / Zip: 10640 N McCarran Bvld. B-109, Reno NV, 89503
Permanent Street / City / State / Zip: 10640 N McCarran Bvld. B-109, Reno NV, 89503
E-mail address: colin83089@yahoo.com
Home phone: 775-720-9543
Application
Cell phone: 775-720-9543
Deadline
March 16, 2012 Acceptance of this award may impact a student's income level to a degree that could affect eligibility for
student loans, and other scholarship/fellowship opportunities. If an applicant has any concerns in this area,
Completed application it is the student's duty to consult with a campus financial aid advisor.
along with reference
letters and transcripts Are you a US citizen or
permanent legal resident alien? Yes No
must be received by
Jessica Bettencourt Do you have a valid US
jbettencourt@unr.edu social security number? Yes No
Are you applying for other
summer research programs? Yes No
Application and reference
letters must be submitted If so, which program(s): NSF EPSCoR Summer UROP
electronically.
Information below is for requested for statistical data only:
High school attended: Homeschool
City/State of high school: Kihei, HI
Date of high school graduation: June, 2007
Birthdate (Month/Day/Year): 08/30/1989
Gender: Female Male

Ethnicity: Hispanic or Latino Not Hispanic or Latino

Race: American Indian/Alaska Native

Asian
Native Hawaiian or Other Pacific Islander
Black or African American
White
More than one race
Disabled: Yes No

I have previously participated in: INBRE UROP Year Pipeline Bridge Workshop Year

Nevada INBRE 2012 UROP Application / 1

 I am currently enrolled at: University of Nevada, Reno
Major: Biology
Number of credits earned by 5/2012: 105
Anticipated graduation date: May, 2013

Reference letters

Two letters of reference (a third is optional) must be emailed directly from the letter's author. One letter
must be from your potential mentor. The other reference(s) should be from someone aware of your
science interests and skills (e.g., a professor from one of your classes.)

Mentor / Reference #1
Name: Dr. Grant Mastick
Email: gmastick@unr.edu
Phone: 775-784-6356
Mentor’s institution: UNR UNLV NVCI UNSOM DRI

Reference #2
Name: Dr. Thomas Kidd
Email: tkidd@unr.edu
Phone: 775-784-6669

Reference #3 (optional)
Name:
Email:
Phone:

Transcripts
Official transcripts or computer generated transcripts must also be received by March 16, 2012. We do
not accept Xeroxed or faxed copies for legibility reasons. If your current institution does not allow you to
download a PDF file of your unofficial transcripts that you can attach to an email, we will accept official
hard-copies that are hand delivered or mailed. If you plan to provide a hard-copy original, please make
your request to the Registrar’s Office 2-3 weeks early. In the past, students have waited until the last
minute and missed our deadline
Transcripts from UNR, TMCC and WNC are attached.

Nevada INBRE 2012 UROP Application / 2

Personal statement
Please describe a) your academic background, b) scientific interests, c) any past research experience, and d) what you hope to gain from the
research experience. Please do not exceed one page.

My college career began at Western Nevada College shortly after I moved to Nevada from my home state of Hawaii. I took
classes that I could transfer to UNR to obtain a Biology degree, while working 40 hours a week at a local skilled nursing
facility to pay for school and my living expenses. Between the lower division classes that I was taking, and experiences in
the field of health care, I soon realized that I wanted to pursue a career in medicine. I was accepted as a transfer student to
UNR in Spring 2011, and quit my job of almost two years to move to Reno and dedicate myself to full time schooling. Since
then, I have received my AS from TMCC, graduating with honors, and have begun undergraduate research on campus at
UNR. My college career has evidenced my diligence and passion for research and medicine, as I have balanced scholastic
endeavors with maintaining a healthy personal life.
As I have progressed through college I have become increasingly interested in the connections between biomedical
research and patient care. Lab and lecture courses have both introduced me to research techniques and the experimental
process. I have been particularly fascinated by the relationship this research has to treating human disease. I see that a
solid understanding of how relevant research is applied to medical applications is vital to a broader understanding of
medicine. On a practical level, my job in a skilled nursing facility provided exposure to patients with diagnoses of
Parkinson’s disease, Alzheimer’s disease and stroke. This experience combined with an elective neurobiology course that I
took last semester provoked me to pursue undergraduate research in the field of neuroscience. When I was offered an
opportunity to begin research in Dr. Mastick’s developmental neurobiology lab this past winter break, I knew that this was
the opportunity I was looking for.
In the two and a half months that I have been in the Mastick lab, I have gained a good level of autonomy, designing
and running experiments on my own. I have become proficient at genotyping with PCR, cyrosectioning samples,
immunohistochemistry, and fluorescent imaging. More recently I have been dissecting mouse embryos for use in whole
mount labeling. I am applying these techniques to experiments involving the positioning of motor neuron cell bodies and
path finding of motor axons in developing mouse embryos. I very much enjoy the independence of working on projects and
providing novel evidence that can be translated into relevant medical applications.
The Summer INBRE grant would allow me to devote more time to the research that I enjoy during the summer
months. If I receive this grant, I am confident that I can make both a contribution to the scientific community, and to my
personal career, by honing techniques I have already begun to use and growing in my independence in the lab. I am
currently planning on applying to medical school, either this summer or the following year, and this experience, along with
the accolade associated with it, will help to solidify that path for my future.

Nevada INBRE 2012 UROP Application / 3

Project proposal
Please create a brief research proposal that is approved by the faculty mentor. The proposal should be a concise statement including a clear
hypotheses to be tested or questions to be asked. Please do not exceed two pages.
Project proposal and references are attached.

Nevada INBRE 2012 UROP Application / 4

Grant Proposal: Floor plate signals position motor neurons in the spinal cord
Author: Colin Wilhelm; Mentor: Grant Mastick, Biology Department, UNR.

Abstract
Guidance cues, specifically those found in the floor plate of the neural tube, play distinct
roles in guiding motor axons to grow out of the spinal cord. While their importance in regulating
motor axon guidance has been clearly elucidated, the function of floor plate signals in the
settlement of motor neuron cell bodies is still unclear. I will test if mutations in the genes
responsible for the floor plate-derived Slit repellent signals, and their Robo receptors, will result
in the abnormal placement of spinal cord motor neurons outside of the neural tube. This project
will determine the role of floor plate signals in maintaining the positioning of motor neuron cell
bodies inside the spinal cord.

Introduction
During embryo development, motor neurons differentiate and settle in a ventral position
near the floor plate (Sabharwal et al., 2011). Spinal cord motor neurons are segregated into
distinct motor pools (Price et al., 2002). These motor pools match with specific muscles at each
spinal level. Proper placement of cell bodies along the spinal cord is vital for the function of this
highly specific pathway (Jessel et al., 2011).
Floor plate signals play an important role in regulating motor axon guidance (Bai et al.,
2011; Bonanomi et al., 2012). In motor axon guidance both Slit secreted repellent proteins, and
their associated Robo receptors are required to repel axons away to their exit point. We have
begun to explore whether floor plate signals also regulate the positioning of motor neuron cell
bodies. Our lab found that mutations in Robo receptors lead to abnormal positioning of motor
neuron cell bodies. In these mutant embryos, some motor neuron cell bodies migrate outside of
the spinal cord, near the motor exit point. This suggests that floor plate signals are required for
the proper placement of motor neuron cell bodies.
Boundary cap cells were previously identified as important in regulating the positioning
of motor neuron cell bodies. Normally, boundary cap cells are precisely located at the motor exit
point to prevent the emergence of motor neuron cell bodies, while allowing the passage of axons
out of the neural tube (Bron et al., 2007). Our new finding in Robo mutant embryos suggests that
Slit repellents are required for the positioning or function of boundary cap cells, which in turn
regulate motor neuron cell body positioning.

Objectives
Aim 1: Determine if motor neuron cell bodies emigrate in Slit1-/-;2-/- and Robo1-/-;2-/- mutant
mouse embryos and quantify the migration phenotype at multiple spinal levels.
Aim 2: Determine if specific classes of motor neurons (eg.LMCl, LMCm, MMC) emigrate in
guidance cue/receptor knockouts.
Aim 3: Explore the effects of guidance cue/receptor mutations on the presence and positioning of
boundary cap cells.

Research Hypotheses
Slit/Robo guidance cues and receptors are individually necessary for the settlement of
motor neuron cell bodies in the spinal cord. The knockout of any one of these cues or receptors
will result in the disruption of boundary cells at the motor exit point, which leads to the
phenotype of emigrant motor neuron cell bodies.

Experimental Plans
Aim 1 – Determine location of cell bodies at multiple spinal levels in knockout embryos
First, I will collect Slit1-/-;2-/- and Robo1-/-;2-/- mouse embryos from mutant lines available
in the Mastick lab, along with wildtype embryos serving as negative controls. These embryos
will be at day 11 of development and I will use PCR to verify their genotypes. Next I will
collect spinal cord sections and perform primary antibody labeling with anti-Islet1 to identify
motor neurons, and anti-beta III-tubulin to show exiting motor nerves (Geisen et al., 2008).
Secondary antibody labeling will be performed with specific fluorescent dye tags and I will
capture images of different spinal levels. I will then quantify the emigrant phenotype in each
kind of mutant by counting the number of emigrating neurons at specific spinal levels and
compare this to wildtype controls. This experiment will confirm the necessity of Slit/Robo in the
regulation of motor neuron cell body position.
Aim 2 – Determine if specific motor neuron classes are more affected by knockouts
For these experiments, I will use sections from the same spinal cords, but label them with
antibodies for specific subtypes of motor neurons. I will employ three primary antibodies, using
anti-Lim3 to identify MMC motor neurons, anti-Lim1 for LMCl identification and anti-Isl1 to
identify all three motor neuron types (Price and Briscoe, 2004). Secondary antibody labeling
with fluorescent tags specific to each primary label will allow for the determination of each
neuron type as a third class of motor neuron (LMCm) will only display Islet1 labeling. Motor
neuron classes will be identified and the number of emigrant cell bodies from each class will be
quantified. This experiment will help to determine if there is a particular kind of motor neuron
that tends to migrate out of the spinal cord.
Aim 3 – Determine the positioning and density of boundary cap cells in knockout embryos
I will collect the same set of embryos as in the previous experiment and perform in situ
hybridization with a Krox20 probe. This transcription factor is a molecular marker in boundary
cap cells (Coulpier et al., 2009). Additionally, I will perform fluorescence antibody labeling
with an anti-Krox20 primary antibody. By using the labels provided by mRNA and antibodies I
can compare the location and quantity of boundary cap cells in mutant and wildtype embryos.
This experiment will show the effects of floor plate signal knockouts on the presence and
positioning of boundary cap cells.

Timetable
Beginning on June 4th, I will complete the collection of embryos for Aims 1-3 by June
th
17 . I will then begin cryosectioning, immunohistochemistry, and in situ hybridization
immediately and will have imaging of these samples completed by July 29th. I will then compile
my data and create a poster for presentation by August 10th.

Dissemination of Results
I will present a poster that summarizes the results of experimentation at the
Undergraduate Summer Research Poster Conference on August 11, 2012. My results will be
organized into figures, which will be included in a manuscript that will be submitted by my
faculty mentor to a peer reviewed journal.
References
Bai, G., O. Chivatakarn, D. Bonanomi, K. Lettieri, L. Franco, C. Xia, E. Stein, L. Ma, J. W.
Lewcock, and S. L. Pfaff. "Presenilin-Dependent Receptor Processing Is Required for
Axon Guidance." Cell 144, no. 1 (2011): 106-18.
<http://www.ncbi.nlm.nih.gov/pubmed/21215373>
Bonanomi, D., O. Chivatakarn, G. Bai, H. Abdesselem, K. Lettieri, T. Marquardt, B. A.
Pierchala, and S. L. Pfaff. "Ret Is a Multifunctional Coreceptor That Integrates
Diffusible- and Contact-Axon Guidance Signals." Cell 148, no. 3 (2012): 568-82.
<http://www.ncbi.nlm.nih.gov/pubmed/22304922>
Bron, R., M. Vermeren, N. Kokot, W. Andrews, G. E. Little, K. J. Mitchell, and J. Cohen.
"Boundary Cap Cells Constrain Spinal Motor Neuron Somal Migration at Motor Exit
Points by a Semaphorin-Plexin Mechanism." Neural Dev 2 (2007): 21.
<http://www.ncbi.nlm.nih.gov/pubmed/17971221>
Coulpier, F., S. Le Crom, G. S. Maro, J. Manent, M. Giovannini, Z. Maciorowski, A. Fischer, M.
Gessler, P. Charnay, and P. Topilko. "Novel Features of Boundary Cap Cells Revealed
by the Analysis of Newly Identified Molecular Markers." Glia 57, no. 13 (2009): 1450-7.
<http://www.ncbi.nlm.nih.gov/pubmed/19243017>
Geisen, M. J., T. Di Meglio, M. Pasqualetti, S. Ducret, J. F. Brunet, A. Chedotal, and F. M. Rijli.
"Hox Paralog Group 2 Genes Control the Migration of Mouse Pontine Neurons through
Slit-Robo Signaling." PLoS Biol 6, no. 6 (2008): e142.
<http://www.ncbi.nlm.nih.gov/pubmed/18547144>
Jessell, T. M., G. Sormeli, and J. S. Kelly. "Motor Neurons and the Sense of Place." Neuron 72,
no. 3 (2011): 419-24. <http://www.ncbi.nlm.nih.gov/pubmed/22078502>
Price, S. R., N. V. De Marco Garcia, B. Ranscht, and T. M. Jessell. "Regulation of Motor Neuron
Pool Sorting by Differential Expression of Type Ii Cadherins." Cell 109, no. 2 (2002):
205-16. <http://www.ncbi.nlm.nih.gov/pubmed/12007407>
Sabharwal, P., C. Lee, S. Park, M. Rao, and S. Sockanathan. "Gde2 Regulates Subtype-Specific
Motor Neuron Generation through Inhibition of Notch Signaling." Neuron 71, no. 6
(2011): 1058-70. <http://www.ncbi.nlm.nih.gov/pubmed/21943603>
 Page 1 of 1
Western Nevada College 11/18/2011
WNC Unofficial Transcripts

Name: Wilhelm,Colin Riley

Former Name: Wilhelm,Colin Riley
Student ID: 1004435184
DOB: 08/30/1989

Beginning of Undergraduate Record

2009 Sumr
Course Description Att. Earned Grade Points
ENG 101 Composition I 3.00 3.00 A 12.000
MATH 96 Intermediate Algebra 3.00 3.00 A 12.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 6.00 6.00 6.000 24.000

2009 Fall
Course Description Att. Earned Grade Points
MATH 128 Precalculus/Trigonometry 5.00 5.00 A 20.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 5.00 5.00 5.000 20.000

2010 Sprg
Course Description Att. Earned Grade Points
CH 201 Ancient & Medieval Cultur 3.00 3.00 A 12.000
CHEM 121 General Chemistry I 4.00 4.00 A 16.000
MATH 181 Calculus I 4.00 4.00 A 16.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 11.00 11.00 11.000 44.000

2010 Sumr
Course Description Att. Earned Grade Points
BIOL 223 Human Anatomy & Phys I 4.00 4.00 A 16.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 4.00 4.00 4.000 16.000

2010 Fall
Course Description Att. Earned Grade Points
BIOL 224 Human Anatomy & Phys II 4.00 4.00 A 16.000
CH 202 The Modern World 3.00 3.00 A 12.000
ENG 102 Composition II 3.00 3.00 A 12.000
STAT 152 Intro to Statistics 3.00 3.00 A 12.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 13.00 13.00 13.000 52.000

Term Honor: Dean's Honors List

Undergraduate Career Totals

Cum GPA: 4.00 Cum Tot 39.00 39.00 39.000 156.000

End of WNC Unofficial Transcripts

 Page 1 of 1
Truckee Meadows Community College 11/18/2011
7000 Dandini Blvd.
Reno, NV 89512
TMCC Unofficial Transcript

Name: Wilhelm,Colin Riley

Former Name: Wilhelm,Colin Riley
Student ID: 1004435184
DOB: 08/30/1989

End of TMCC Unofficial Transcript

Degrees Awarded
Degree: Associate of Science
Confer Date: 05/15/2011
Degree Honors: Degree Honors with Distinction
Plan: ASSOCIATE OF SCIENCE, BIOLOGY
_____________________________

Test Credits

Test Credits
Att. Earned Points
Test Trans GPA: 0.00 Transfer Tot: 3.00 3.000 0.000

Transfer Credits
Transfer Credit from Western Nevada College
Att. Earned Points
Course Tran GPA: 0.00 Transfer Tot: 36.00 36.00 0.000

Beginning of Undergraduate Record

2010 Spring
Course Description Att. Earned Grade Points
BIOL 190 Intro Cell/Molecular Biol 3.00 3.00 A 12.000
BIOL 190L Cell/Molecular Biol Lab 1.00 1.00 A 4.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 4.00 4.00 4.000 16.000

2010 Summer
Course Description Att. Earned Grade Points
MUS 121 Music Appreciation 3.00 3.00 A 12.000
PSY 101 General Psychology 3.00 3.00 A 12.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 6.00 6.00 6.000 24.000

2010 Fall
Course Description Att. Earned Grade Points
BIOL 191 Intro Organismal Biology 3.00 3.00 A 12.000
BIOL 191L Intro Organismal Biol Lab 1.00 1.00 A 4.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 4.00 4.00 4.000 16.000

2011 Spring
Course Description Att. Earned Grade Points
CH 203 Am Exp & Const Change 3.00 3.00 A 12.000
CHEM 122 General Chemistry II 4.00 4.00 A 16.000
PHIL 210 World Religions 3.00 3.00 A 12.000
PHYS 151 General Physics I 4.00 4.00 A 16.000
Att. Earned GPA Unit Points
Term GPA 4.00 Term Tot 14.00 14.00 14.000 56.000

2012 Spring
Course Description Att. Earned Grade Points
BIOL 251 General Microbiology 4.00 0.00 0.000
NUTR 223 Prin of Nutrition 3.00 0.00 0.000
Att. Earned GPA Unit Points
Term GPA 0.00 Term Tot 7.00 0.00 0.000 0.000

Undergraduate Career Totals

Cum GPA: 4.00 Cum Tot 35.00 28.00 28.000 112.000
Wilhelm,Colin Riley University of Nevada, Reno
2165 Carville Rd. #11 H 1664 N Virginia St
Reno, NV 89512 Reno, NV 89557

Student ID: 1004435184 Page 1 of 1

Birth Date: 08/30/1989 Unofficial Transcript Print Date: 01/14/2012

Transfer Credits
Attempt Earned GPA Pt GPA
Truckee Meadows 31.0 28.0 112.0 4.000
Community College

Western Nevada College 36.0 36.0 144.0 4.000

Beginning of Undergraduate Record

2011 Spring

BIOL 300 Principles of Genetics 3.0 3.0 A

Attempt Earned GPA Crd GPA Pt GPA

Term 3.0 3.0 3.0 12.0 4.000
Career 3.0 3.0 3.0 12.0 4.000

2011 Fall

BIOL 315R Cell Biology 3.0 3.0 A

BIOL 316 Compar Animal Phys 3.0 3.0 A
BIOL 395 Lab Genetic/Cell Biol 2.0 2.0 A
BIOL 475 Neurobiology 3.0 3.0 B+
BIOL 477 Genes, Brain & Behavior 3.0 3.0 A-
CHEM 341 Organ Chem Sci Prof I 3.0 3.0 C
HDFS 201 Lifespan Human Develop 3.0 3.0 A

Attempt Earned GPA Crd GPA Pt GPA

Term 20.0 20.0 20.0 71.0 3.550
Career 23.0 23.0 23.0 83.0 3.609

2012 Spring

BCH 400 Intro Biochem 4.0 0.0

BIOL 415 Evolution 4.0 0.0
CHEM 342 Organic Chem Sci Prof II 3.0 0.0
PSY 441 Abnormal Psychology 3.0 0.0

Attempt Earned GPA Crd GPA Pt GPA

Term 14.0 0.0 0.0 0.0 0.000
Career 37.0 23.0 23.0 83.0 3.609

End of Undergraduate Record

Under the provisions of the Family Educational Rights and Privacy Act of 1974, as amended, this record is not
to be released to a third party without the written consent of the student.