Cell-Matrix Adhesion in Muscle Development and Disease

210 Pages·2016·8.72 MB·English

The University of Maine
DigitalCommons@UMaine
Electronic Theses and Dissertations Fogler Library
8-2012
Cell-Matrix Adhesion in Muscle Development and
Disease
Michelle F. Goody
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Recommended Citation
Goody, Michelle F., "Cell-Matrix Adhesion in Muscle Development and Disease" (2012).Electronic Theses and Dissertations. 1801.
http://digitalcommons.library.umaine.edu/etd/1801
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CELL-MATRIX ADHESION IN MUSCLE
DEVELOPMENT AND DISEASE
by
Michelle F. Goody
B.S. The University of Maine, 2007
A DISSERTATION
Submitted in Partial Fulfillment of the
Requirements for the Degree of
Doctor of Philosophy
(in Biomedical Sciences)
The Graduate School
The University of Maine
August, 2012
Advisory Committee:
Clarissa Henry, Associate Professor, School of Biology and Ecology, University
of Maine, Advisor
Dorothy E. Croall, Professor of Biochemistry, Department of Molecular and
Biomedical Sciences, University of Maine
Leif Oxburgh, Principal Investigator, Maine Medical Center Research Institute
Roger Sher, Associate Research Scientist, The Jackson Laboratory
Mary Tyler, Professor, School of Biology and Ecology, University of Maine
DISSERTATION ACCEPTANCE STATEMENT
On behalf of the Graduate Committee for Michelle F. Goody I affirm that this
manuscript is the final and accepted dissertation. Signatures of all committee members
are on file with the Graduate School at the University of Maine, 42 Stodder Hall, Orono,
Maine.
Clarissa Henry, PhD, Advisor Date
Copyright Michelle F. Goody 2012
iii
LIBRARY RIGHTS STATEMENT
In presenting this dissertation in partial fulfillment of the requirements for an
advanced degree at the University of Maine, I agree that the Library shall make it freely
available for inspection. I further agree that permission for “fair use” copying of this
dissertation for scholarly purposes may be granted by the Librarian. It is understood that
any copying or publication of this dissertation for financial gain shall not be allowed
without my written permission.
Signature:
Date:
CELL-MATRIX ADHESION IN MUSCLE
DEVELOPMENT AND DISEASE
By Michelle F. Goody
Advisor: Dr. Clarissa Henry
An Abstract of the Dissertation Presented
in Partial Fulfillment of the Requirements for the
Degree of Doctor of Philosophy
(in Biomedical Sciences)
August, 2012
A variety of diseases, both inherited and acquired, affect muscle tissues in
humans. The anchoring of muscle fibers to their surrounding environment is critical for
muscle homeostasis. Muscle fibers attach to their microenvironment through cell-matrix
adhesion complexes. These anchoring complexes are placed under repeated stress during
muscle contraction. Genetic mutations in these complexes weaken the attachment
between muscle fibers and their microenvironment, making fibers more susceptible to
damage and death. This increased fiber degeneration eventually leads to progressive
muscle wasting diseases, known as congenital muscular dystrophies. Although clinical
trials are ongoing, there is presently no way to cure the loss of muscle structure and
function associated with congenital muscular dystrophies.
Animal models of human diseases are used to gain insights into mechanisms of
disease pathogenesis and to screen for potential therapeutic compounds. The zebrafish
model system, well-known for its use in developmental biological studies, is rapidly
becoming widely-accepted as a useful model for biomedical research. We utilized
zebrafish embryos to study the initial morphogenesis of substructures in the muscle
microenvironment and the initial stable cell-matrix adhesions formed in muscle tissue. As
the muscle fiber microenvironment is abnormal in congenital muscular dystrophies and
cell-matrix adhesions are weakened, studies elucidating how strong, stable cell-matrix
adhesions form in development could be informative in the effort to treat congenital
muscular dystrophies.
Using this approach, we identified a previously undescribed cell-matrix adhesion
pathway required for normal organization of an important substructure in the muscle
tissue microenvironment. We show that activation of this cell-matrix adhesion pathway in
dystrophic zebrafish not only significantly reduces muscle degeneration, but also
improves swimming ability. The results presented in this dissertation identify proteins
that function in this cell-matrix adhesion pathway and use dystrophic zebrafish to show
the benefits and limitations of this pathway in treating symptoms of congenital muscular
dystrophies. Our findings suggest that activation of this pathway has the potential to
ameliorate loss of muscle structure and function in multiple muscular dystrophies.
ACKNOWLEDGEMENTS
Thank you to my committee members, Drs. Croall, Oxburgh, Sher and Tyler, for
your perspectives, feedback and guidance during my graduate education and graduate
research experience. I would especially like to thank Mary Tyler. Mary - not only was it a
pleasure to be your student and learn from you, but your recommendation to introduce
myself to Dr. Clarissa Henry changed the path that I was on and resulted in me
embarking on the adventure of graduate school.
I can only begin to express my gratitude for my advisor, Dr. Clarissa Henry.
Clarissa - it was an absolute joy to be trained by you and to conduct my graduate research
project in your lab under your guidance. I feel the top-notch, well-rounded training that I
received from you at UMaine has provided me with a solid foundation to competitively
pursue any science-related career. I am thankful for the opportunities that you afforded
me to travel to scientific conferences and for teaching me how to effectively
communicate my work. The atmosphere in your lab, an ideal balance between
productivity and fun, made me motivated and genuinely excited to be at the lab each day.
I would like to thank all the members of the Henry lab that I got to work with,
past and present. Thanks to a wonderful technician, Mary Astumian, for experimental
help as well as keeping the day-to-day operations of the lab running smoothly. Thank you
to former Henry lab graduate students Matt Peterson, Molly Jenkins and Erik McCarthy
for collaborations and helpful discussions of my project (especially Erik for piecing
together that MIBP2 and Nrk2b are the same protein, which resulted in a whole new
direction to explore). I would like to thank the outstanding undergraduate researchers that
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I got the opportunity to work with and help train, including Judi Azevedo, Kevin Lessard,
Sam Entwisle, Christine Reynolds and Anna Burgess. Also, thanks to Cindi Gousse, an
undergraduate worker, for doing the unglorified, but necessary chores that keep a lab
running. I owe a debt of gratitude to Meghan Kelly. Meghan - my time in graduate school
was so enjoyable in large part because you were right there next to me, whether we were
in class, at a conference or seminar, writing at our desks, injecting embryos, imaging at
the microscopes, cross-country skiing or at the movie theatre. Your input and support,
professionally and personally, are invaluable to me and I am truly grateful that graduate
school introduced us to one another.
I found a home in UMaine’s Graduate School of Biomedical Sciences and would
like to thank the GSBS director Dr. Carol Kim. Carol - you have put so much thought and
effort into making the GSBS a student-centered, intellectually stimulating, collaborative
and fun experience and you have succeeded in your vision. Also, thanks to my fellow
GSBS peers and members of nearby laboratories, especially Kim Brothers, Dr. Remi
Gratacap, Kristin Gabor, Jeremy Charette and Rachel Palmer, for helpful advice,
discussions of science and fun times.
I would like to thank UMaine’s Thesis Studio co-founder, Dr. Dylan Dryer, and
the members of my particular Thesis Studio, especially Robert Hodges, for making the
writing of my dissertation enjoyable. It was a wonderful experience to receive helpful
critiques of my writing and to be able to help others improve their writing.
Thanks to Mark Nilan for outstanding care and maintenance of the UMaine
zebrafish facility. The health and fecundity of the zebrafish at UMaine are unparalleled in
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large part to your ingenuity and meticulousness; the value of this resource is not taken for
granted.
Lastly, I would like to thank my extremely supportive family. Thank you to my
parents for the importance you place on education and for loving and supporting me in
everything I do, to my sisters for being my best friends, and to my husband, Brad, and his
family for constant love and encouragement. Thank you Brad for all the times that you
have listened to me explain what I did at work that day and waited in the car for me while
I just had to run into the lab for “five” minutes. Brad, I have loved every minute of
raising our five, furry, four-legged children with you and can not wait to start raising our
daughter together very soon; your personality is the perfect compliment to mine and your
humor and outlook remind me what is important in life. Thank you!
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