PSL Rop 2012-2013

RESEARCH OPPORTUNITY PROGRAM
299Y PROJECT DESCRIPTIONS 2012‐2013
FALL/WINTER Project Code: PSL 1

Name and Title: Prof. S. Lee Adamson
Department: Obstetrics & Gynaecology, and Physiology
Phone Number: 416‐586‐8377 Email: adamson@lunenfeld.ca

TITLE OF RESEARCH PROJECT: Interactions between the Placenta and the Mother in Determining
Pregnancy Outcomes in Mice.

NUMBER OF STUDENT PLACES AVAILABLE: 2

OBJECTIVES AND METHODOLOGY: Our lab is using genetically altered mice to examine the effect of
mutations in angiogenic genes in the placenta on maternal and fetal outcomes in pregnancy. In most
models, genetic mutations are only in the fetal trophoblast cells that form the placenta and only half the
fetuses in the litter are affected. Nevertheless, changes in maternal function in the wildtype mothers
and in their wildtype littermates have been observed revealing the importance of placental‐maternal
interactions in pregnancy. Our current objectives are to determine the effects of altering the VEGF
angiogenic pathway in trophoblast on placental morphology and vascularity. Vascularity in placentas is
being quantified using histomorphometry image analysis software, and by visualizing plastic or rubber‐
filled placental vasculature using scanning electron microscopy and micro‐computed tomography
imaging.

DESCRIPTION OF STUDENT PARTICIPATION: Students will participate in collection and analysis of
placental samples with the assistance of technical staff in the lab. They will learn placental anatomy,
how the placenta interacts with maternal physiology to affect placental morphology and vascularity, and
will learn state‐of‐the‐art methods for tissue and vascular imaging and quantification. They will perform
quantitative image analysis and be responsible for collating and statistically analyzing their data. They
will also collect high quality images to supplement their findings, and will participate in lab events and
lab meetings when feasible given their academic schedule.

MARKING SCHEME (assignments with weight and due date):
 10%‐ 2‐page report due 15 November (double‐spaced, including rationale of project, hypothesis,
methods)
 30% ‐ final report (~10 pages double‐spaced, including introduction, methods, results and
discussion and references, in standard journal format; figures and legends should be included,
but do not count towards the page limit; by April 5, 2013)
 10% ‐ Participation in weekly Journal clubs/lab meetings (by April 5, 2013)
 10% ‐ Oral presentation to the lab or poster presentation in Undergraduate Research Fair for
ROP students (held in March) (by April 5, 2013)
 30% ‐ lab mark: 15% by December 31; 15% by April 5, 2013
 10% ‐ Final data and lab books, labeled and explained for lab archives (by April 5, 2013)


Name and Title: Patricia Brubaker, Professor and Canada Research Chair
Department: Physiology
Phone Number: 416‐978‐2593 Email: p.brubaker@utoronto.ca

TITLE OF RESEARCH PROJECT: Synthesis, Secretion and Bioactivities of the Intestinal Glucagon‐like
Peptides


OBJECTIVES AND METHODOLOGY:
Under the direct supervision of both the PI and a senior graduate student/post‐doctoral fellow, the
project student will conduct immunohistochemical analysis of intestinal sections for proteins involved in
the secretion and/or actions of glucagon‐like peptide‐1 and glucagon‐like peptide‐2 – these are peptide
hormones with key roles in the maintenance of glucose and intestinal homeostasis, and that are now
being used clinically for the treatment of diabetes and short bowel syndrome, respectively.

DESCRIPTION OF STUDENT PARTICIPATION:
The student will be responsible for all aspects of the project, from experiments to analysis to data
presentation. The PI will ensure that the student obtains all necessary training in required laboratory
techniques, as well as that the student takes all UofT courses required for any student working in the
laboratory (regardless of the project), including Biosafety and Radiation.

methods)


Name and Title: Zhong‐Ping Feng
Phone Number: (416)946‐0671(office) Email: zp.feng@utoronto.ca

TITLE OF RESEARCH PROJECT: Regulatory Mechanisms of Synaptic Function


OBJECTIVES AND METHODOLOGY: Synaptic activities are essential for neuronal communication and are
regulated during development, learning and memory process or after neuronal injury. The main
objectives of the research projects are to investigate the cellular and molecular mechanisms involved in
the synaptic regulation and subsequence neural behavioural changes using a fresh‐water pond snail,
Lymnaea stagnalis, as animal model. These studies are carried out both in vitro and in vivo systems,
using cell culture, electrophysiological recordings, optical imaging, immunochemical, and molecular
biological techniques.

Students are expected to participate in one of the indicated research projects, understand the concepts
of the project, learn and perform one or two of the following tasks: 1) animal behavioural evaluation 2)
data analysis 3) cell culture 4) optical imaging 5) basic molecular assays 6) basic biochemical analysis

methods)


Name and Title: Dr. Sheena Josselyn, Associate Professor
Department: Physiology, Institute of Medical Sciences
Phone Number: 416‐813‐7654 x 1824 Email: sheena.josselyn@sickkids.ca

TITLE OF RESEARCH PROJECT: Developing a Novel Touchscreen‐Based Task for Modeling Depression in
Mice


This project will involve the development of a novel approach to model depression in mice using
touchscreen‐based operant chambers as well as standard operant chambers. The tasks used in the
touchscreen‐equipped operant chamber will be simultaneous visual discriminations using computer‐
generated achromatic visual stimuli, while tasks used in the standard operant chambers will involve bar
pressing for food pellets and fear‐conditioning to auditory stimuli.

Students will be responsible for training mice daily to perform visual discriminations using touchscreen‐
equipped operant chambers as well as training mice daily to bar press for food pellet rewards using
standard operant chambers. Students will collect and analyze behavioural data. Students will be
responsible for conducting a literature review, creating an annotated bibliography of relevant papers,
and writing a manuscript‐style final report detailed and describing the collected data.

methods)


Name and Title: Fang Liu, Professor
Department: Department of Neuroscience
Phone Number: (416) 535‐8501 ext. 4659 Email: fang_liu@camh.net

TITLE OF RESEARCH PROJECT: Roles of Alpha‐Synuclein and Parkin in Excitotoxicity


Numerous data reveal the involvement of alpha‐synuclein and parkin in dopaminergic neuronal loss, yet
the detailed role of these two molecules in this process is unknown. The research is aimed to study the
roles of alpha‐synuclein and parkin in dopamine‐ and glutamate‐mediated cell death. Using recombinant
cell systems, the relation and contribution of alpha‐synuclein and parkin to cell death process will be
studied.

Students will perform the experiments with established methods, and analyze and report results and
possible problems encountered during experiments. They are also encouraged to from their own
hypothesis with further experimental directions based on the results obtained.

methods)


Name and Title: Jinrong Min, Assistant professor/PI
Department: Department of Physiology/ Structural Genomics Consortium, University of Toronto
Phone Number: 416‐9463868 Email: jr.min@utoronto.ca

TITLE OF RESEARCH PROJECT: Structural Studies of Chromatin Proteins


OBJECTIVES AND METHODOLOGY: Learn basic lab skills in protein biochemistry, and use these skills in
protein purification and crystallization

The students will participate in research projects in my lab to characterize chromatin proteins by X‐ray
crystallography in combination with other biochemical and biophysical techniques. The students will
learn how to prepare the LB media for E Coli cell growth, prepare competent cells and LB agar plates for
transformation, and make different buffers for protein purification. They are also expected to learn the
recombinant DNA technology for subcloning the proteins of interest, and learn protein purification
methods with senior members in the lab. There is also a chance for them to learn how to do protein
crystallization in the end of the course.

methods)


Name and Title: Dr. Beverley A. Orser, MD, PhD, FRCPC, Canada Research Chair in Anesthesia
Department: Physiology and Anesthesia
Phone Number: 416 978 0574 Email: beverley.Orser@utoronto.ca

TITLE OF RESEARCH PROJECT: Memory Loss after Anesthesia


Memory deficits persist after general anesthesia or sedation much longer than expected based on drug
pharmacokinetics. Most anesthetics and sedative drugs increase the activity of inhibitory GABA‐A
receptors. The goal of the project is to: 1) identify the cause of long‐term memory loss after anesthesia
and sedation and 2) determine whether drugs that cause sedation but do not target GABA‐A receptors
(such as the adrenergic agonist dexmedetomidine) also cause memory deficits. Memory will be studied
using mouse models of fear‐associated memory and non‐adversive memory paradigms. We anticipate
that the results from the proposed studies will advance our understanding of the cellular mechanisms
underlying memory and also provide clues to the optimum drug treatment for patients undergoing
anesthesia and sedation.

Students will gain hands‐on experience designing and performing behavioral experiments using several
genetically modified mouse strains. They will participate as a member of a lab team that is focused on
understanding the molecular basis of memory.

methods)


Name and Title: Ian Rogers PhD, Assistant Professor
Department: Obstetrics and Gynecology, Cross appointment in Physiology
Phone Number: 416‐586‐4800 x 4122 Email: rogers@lunenfeld.ca

TITLE OF RESEARCH PROJECT: Bio‐Artificial Organs‐ Stem Cells to Repair Damaged and Diseased Tissues


My lab is focused on developing cell therapies to treat various human diseases. In most cases we use

human stem cells. The diseases we are most interested in are 1) diseases that can be treated by bone
marrow transplants, such as leukemia, 2) type 1 diabetes and 3) peripheral vascular disease. Our goal is
to use stem cells to produce replacement tissues. For example we use umbilical cord blood stem cells to
help us develop more successful regimens for bone marrow transplants. We also use umbilical cord
blood stem cells to produce blood vessels to repair or replace damaged vessels that occur in patients
suffering from peripheral vascular disease.
The student will be supervised by a lab member and taught typical cell biology techniques used in the
lab. These will include cell culture techniques using cell lines. Cell imaging techniques including
immunocytochemistry and immunohistochemistry and other methods of analysis such as PCR will be
taught.
1. 10%‐ 2‐page report due 15 November (double‐spaced, including rationale of project, hypothesis,
methods)
2. 30% ‐ final report (~10 pages double‐spaced, including introduction, methods, results and
3. 10% ‐ Participation in weekly Journal clubs/lab meetings (by April 5, 2013)
4. 10% ‐ Oral presentation to the lab or poster presentation in Undergraduate Research Fair for
5. 30% ‐ lab mark: 15% by December 31; 15% by April 5, 2013
6. 10% ‐ Final data and lab books, labeled and explained for lab archives (by April 5, 2013)


Name and Title: Qinghua Wang, Associate Professor
Department: Physiology and Medicine
Phone Number: (416) 864‐6060 x77610 Email: wangq@smh.ca

TITLE OF RESEARCH PROJECT: Dynamics of Islet Beta‐Cell Mass in the Regulation of Glucose
Homeostasis in Mice.

NUMBER OF STUDENT PLACES AVAILABLE: 1‐2

Loss of islet beta‐cell mass is a course underlying beta‐cell dysfunction and insulin deficiency in both
type 1 and type 2 diabetes. Theo objective if this course is to learn how the islet beta‐cells are regulated
and their contribution to the glycemic control and in the development of diabetes in mouse diabetes
models. The primary methodology is Histochemistry in pancreatic tissue to identify insulin‐ and
glucagon‐secreting cells, and the Beta‐cell Mass Analysis is conducted based on specific computer
software i.e. ImageScope.

The students will learn the procedure of Immunostaining, image capture using microscope, master the
principal and protocol of computer viewing program, and to certain extent, to get familiar with the basic
skill of animal handling, including injections and glucose tolerance test etc.

methods)


Name and Title: Dr Carin Wittnich, Professor
Phone Number: 416‐978‐7744 Email: c.wittnich@utoronto.ca

TITLE OF RESEARCH PROJECT: Role of Heavy Metal Contaminants Ii Marine Mammals and Human Health


Objectives are to clarify levels of heavy metals in fish and marine mammal species found along Canada’s
coastline that also serve as food sources for the indigenous peoples and other local populations.
Methodology includes collecting samples of key tissue such as muscle, blubber, liver, kidney etc taken at
necropsy. The tissue is preserved through freezing and transported to the laboratory at the University of
Toronto. There the tissue is prepared for subsequent analysis for full heavy metal profiles using mass
spectrophotometry.

Student will participate in the collection of the samples; preparation of the tissue in my laboratory at the
medical sciences bldg; analysis of the results; perform statistics; conduct and compile literature etc. For
the 299 student there is no travel involved; I conduct some necropsies on campus and in those the
student would participate in the tissue collection so that they are made aware of this component.
Normally my collaborators from other locations take the samples, freeze them and they are sent to my
laboratory for processing.


• 10%‐ 2‐page report due 15 November (double‐spaced, including rationale of project, hypothesis,
methods)
• 30% ‐ final report (~10 pages double‐spaced, including introduction, methods, results and discussion
and references, in standard journal format; figures and legends should be included, but do not count
towards the page limit; by April 5, 2013)
• 10% ‐ Participation in weekly Journal clubs/lab meetings (by April 5, 2013)
• 10% ‐ Oral presentation to the lab or poster presentation in Undergraduate Research Fair for ROP
students (held in March) (by April 5, 2013)
• 30% ‐ lab mark: 15% by December 31; 15% by April 5, 2013
• 10% ‐ Final data and lab books, labeled and explained for lab archives (by April 5, 2013)

Name and Title: Martin Wojtowicz
Phone Number: 416 978 2899 Email: martin.wojtowicz@utoronto.ca

TITLE OF RESEARCH PROJECT: A Study of Neuronal Plasticity in the Brain


To understand the structural and functional changes in the brains of experimental animals that
accompany learning and memory.

Students will take part in experiments by taking physiological measurements on laboratory rats and
mice. They will learn surgical techniques and preparation of in vitro brain slices. They will perform
fixation of brain tissue for histology and immunohistochemistry. Perform tissue sectioning and staining
for markers of neuronal plasticity.

Students will work under guidance of a senior graduate student, postdoctoral fellow or technician and
will have to commit to spend certain amount of time in the laboratory. Most projects will require
spending 2 half‐days or one full day per week. Careful attention to instructions, meticulous note taking
and record keeping are a must. Dexterity and ability to operate electronic equipment are an asset.
Initiative and ability to read background literature will be welcome.

MARKING SCHEME
methods)


Name and Title: Dr. Mei Zhen, Associate Professor
Phone Number: 416‐586‐1592 Email: zhen@lunenfeld.ca

TITLE OF RESEARCH PROJECT: Determine the Physiological Role of UBR1, a Gene Associated with the
Johanson‐Blizzard Syndrome


Objectives: Missense mutations in the UBR1 gene lead to the Johanson‐Blizzard Syndrome, an
autosomal recessive congenital disorder, with hallmark features in pancreatic exocrine insufficiency and
mental retardation. UBR1 encodes for an E3 ubiquitin ligase, a family of enzymes that selectively target
cellular proteins for degradation. It is hypothesized that in Johanson‐Blizzard Syndrome patients, an
aberrantly high activity of the UBR1‐substrates causes abnormal development/function of the
pancreatic and nervous system. The physiological substrates of UBR1 for both systems, however, remain
completely unknown. The objective of this project is to use an animal model to 1) determine the
physiological role of UBR1, and 2) identify signal pathways through which UBR1 negatively regulates.

Methods: Through a genetic screen designed to identify genes that regulate the development and
function of a neuroendocrine‐like neuron in C. elegans, we recently identified a loss of function
mutation allele of UBR‐1, the sole C. elegans homologue of human UBR1 (Chitturi, Hung and Zhen, in
preparation). We found that in ubr‐1 mutant animals, this neuron fails to differentiate proper release
sites, and exhibits a decreased efficiency in releasing neuropeptides. The C. elegans ubr‐1 mutant
therefore represents an excellent animal model to understand the physiological function of UBR1 and to
identify potential targets of UBR1 relevant for nervous system development and function. We will
pursue these goals through following methodologies:

1) Examine the role of UBR‐1 in nervous system development: Through electron microscopy (EM),
we will trace and reconstruct the cross sections of this neuron to determine the cellular
structural changes in this neuron. Specifically, we will examine the distribution and morphology
of synaptic and dense core vesicles along the axon of the neuron, and compare the difference
between wild‐type and ubr‐1 mutants.

2) Identify molecular components of signaling pathways targeted by UBR1: A proven approach to
identify E3 ubiquitin ligase targets is to identify genetic suppressors of the E3 ligase mutant. In
the absence of the E3 ligase activity, the aberrantly increased target activity leads to the
manifestation of the mutant phenotypes. Therefore, a genetic lesion in the substrate, which
leads to a concurrent decrease of the substrate activity in the E3 ligase mutant background, will
rescue the E3 mutant phenotype. The C. elegans ubr‐1 mutants exhibit reduced growth rate and
sluggish locomotion. We will mutagenize the genome of ubr‐1 mutant, and select for progenies
that exhibit restored locomotion and growth. The precise location of the genetic lesion in these
suppressors will be mapped by standard single nucleotide polymorphism (SNP) mapping,
followed by whole‐genome sequencing, a approach that we have successfully undertaken to
isolate and clone the ubr‐1 mutant in the first place.

Student 1: This student will participate in the EM analysis that is currently led by a Ph. D student
Jyothsna Chitturi in my lab and an EM specialist Doug Homlyard at the Department of Pathology (Mount
Sinai Hospital). Doug and Jyothsna have prepared serial thin sections from the EM samples of wild‐type
and ubr‐1 animals. This student will assist them in obtaining digital images of these sections, reconstruct
and analyze the 3D image of the ultrastructure of this neuron.

Student 2: This student will participate in the genetic suppressor screen of the ubr‐1 mutant that is
aimed at identifying UBR‐1 substrates. This student will be under the direct supervision by both Jyothsna
Chitturi and Wesley Hung (a research associate in my lab), who have identified and cloned the ubr‐1
mutant. They have designed the genetic suppressor screen and cloning strategies of the ubc‐1 mutant.
This student will participate in chemical mutagenesis, mutant screening, genetic crosses, SNP mapping
and DNA sample preparation for whole‐genome sequencing (WGS).

These projects are intent for undergraduate students who are considering graduate studies to obtain
training and experience in research. Therefore, both students will attend our weekly lab meeting/journal
club where they will learn the fundamentals about scientific research. They are expected to do two oral
presentations. In Dec., 2012, they will each give a presentation that focuses on the background and
methodology review on their projects. In March 2013 (at the end of their course), they will give a final
presentation that consists backgrounds and results on their projects.

methods)

Name and Title: Min Zhuo, RSC, Professor and Canada Research Chair
Phone Number: 416‐978‐4018 Email: min.zhuo@utoronto.ca

TITLE OF RESEARCH PROJECT: Mapping Cortical Plasticity using MED64 Recording System

NUMBER OF STUDENT PLACES AVAILABLE: Two (2)

Under the direct supervision of both the PI and a senior graduate student/post‐doctoral fellow, the
project student will conduct electrophysiological experiments to study cortical plasticity (long‐term
potentiation (LTP) and long‐term depression (LTD)) in mouse in vitro brain slices. Key genetic
manipulated mice will be used to investigate molecular mechanisms of memory and pain related
plasticity.

The student will be responsible for all aspects of the experiments, from learning the preparation of brain
slices, the use of electrophysiological equipments, to analysis to data presentation. The PI will ensure
that the student obtains all necessary training in required laboratory techniques, as well as that the
student takes all UofT courses required for any student working in the laboratory (regardless of the
project), including Biosafety and Radiation.

methods)

PSL Rop 2012-2013

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RESEARCH OPPORTUNITY PROGRAM

My lab is focused on developing cell therapies to treat various human diseases. In most cases we use

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