1. RESEARCH POSTER PRESENTATION DESIGN © 2012
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Intermediate filaments (IFs) serve essential functions as
cytoskeletal components, providing structural support to
cellular bodies and regulating mechanical stress. Deficiencies in
IF structure, hence function, lead to a variety of conditions,
most significantly desmin-related myopathy. However, such
conditions rarely result from qualitative defects alone, but
emerge in combination with quantitative losses in the amount
of IF expressed. Previously, Caenorhabditis elegans was used as
a model organism to establish that mechanical stress reduction
through two methods, immersion in a soft agar environment and
introducing a separate mutation that restricts locomotion, both
effectively reversed qualitative defects. The aim of this project
was to reexamine those stress reduction methods while
mimicking quantitative IF deficiencies. Because both previous
stress reduction methods were immersive in nature, it was
hypothesized that the treatments would be equally efficacious
when a quantitative mutation was introduced. To simulate such
defects, the lin-14 heterochronic gene was utilized to shift
developmental patterns of IF quantity. Its impact was observed
in combination with structural defects in IFA-2 and IFA-3. The
previous stress reduction techniques were imposed upon
populations with qualitative and quantitative defects. Both
methods of stress reduction were equally effective with the
addition of the quantitative deficiencies, supporting the initial
hypothesis.
Abstract
Introduction
The problem today is there limited research into the therapeutic treatments for conditions
available for Desmin-Related Myopathy, as well as similar conditions related to IF deficiencies.
The purpose was to establish the baseline theory of stress reduction as a applicable form of
treatment for IF deficiencies like DRM
Problem/Purpose Results
Conclusion
The results supported the initial
hypothesis
Outlays and confirms theory of stress
reduction
Therapeutic Stress Reduction
• Quantitative alterations did not decrease
effectiveness
• Both are immersive in nature
• Future studies could extend to DRM or other
similar conditions
• Stress reduction methods as applicable to
humans
References
• Altun, Z.F., & Hall, D.H. (2009). Introduction to C. elegans anatomy. In WormAtlas. Retrieved from
http://wormatlas.org/hermaphrodite/introduction/Introframeset.html
• Ambros, V. (1997). C. elegans II. 2nd edition. New York, NY: Cold Spring Harbor Laboratory Press. Retrieved from
http://www.ncbi.nlm.nih.gov/books/NBK20033/?report=reader#!po=25.0000
• Cooper, G.M. (2000). The cell: A molecular approach. 2nd edition. Sunderland, MA: Sinauer Associates. Retrieved from
http://www.ncbi.nlm.nih.gov/books/NBK9834/
• Goldfarb, L.G., Vicart, P., Goebel, H.H., & Dalakas, M.C. (2004). Desmin myopathy. Brain, 127, 723-724. doi:
10.1093/brain/awh033
• Hodgkin, J., Horvitz, H.R., & Brenner, S. (1979). Nondisjunction mutants of the nematode Caenorhabditis elegans. Genetics,
91, 67-94.
• Horvitz, H.R. & Sulston, J. E. (1980). Isolation and genetic characterization of cell-lineage mutants of the nematode
Caenorhabditis elegans. Genetics, 96, 435-454.
• Karabinos, A., Schmidt, H., Harboth, J., Schnabel, R., & Weber, K. (2001) Essential roles for four cytoplasmic intermediate
filament proteins in Caenorhabditis elegans development. Proceedings of the National Academy of Sciences, 98, 7863-7868.
doi: 10.103/pnas.121169998
• Karabinos, A., Schulze, E., Schunemann, J., Parry, D.A.D., & Weber, K. (2003) In vivo and in vitro evidence that the four
essential intermediate filament (IF) proteins A1, A2, A3 and B1 of the nematode Caenorhabditis elegans form an Obligate
Heteropolymeric IF system. Journal of Molecular Biology, 333, 307-319. doi: 10.1016/j.jmb.2003.08.041
• Labouesse, M. (2006). Epithelial junctions and attachments. Retrieved from
http://www.ncbi.nlm.nih.gov/books/NBK19677/
• Lodish, H., Berk, A., Zipursky, S.L., et. al. (2000). Molecular cell biology. 4th edition. New York: W. H. Freeman. Available
from http://www.ncbi.nlm.nih.gov/books/NBK21560/
• Rutgers University. (n.d.). C. elegans as a Model System. Retrieved from
http://avery.rutgers.edu/WSSP/StudentScholars/project/introduction/worms.html
• Saito, R.M., & van den Heuvel, S. (2002). Malignant worms: What cancer research can learn from C. elegans [Abstract].
Cancer Investigations, 20(2), 264-275.
• Zhu, E. (2014). Examining Stress Reduction on mua-6 and ifb-1 Intermediate Filament Weak Points in C. elegans.
Unpublished document.
Goldfarb, et al., 2004
• Desmin-Related Myopathy centers on genetically-
inherited deterioration of desmin
• Current treatments are nonexistent
Lodish, et al., 2000
• IFs are compromised of 3 primary components
• Universal structure and function of providing
mechanical support
Hapiak, et al., 2003
• Epidermal IFs expressed in Fibrous Organelle (FO)
• IFA-2, IFA-3, IFB-1
By: Eric Zhu (Ohio)
Dr. John Plenefisch of the University of Toledo Dept of Biological Sciences and Ms. Blythe Tipping of Sylvania Southview
High School
Temporal and Osmotic Regulation of Intermediate Filament Function in the C. elegans Epidermis
Figure 1
Figure 2
Hypothesis
Decreasing stress on mua-6 and 1fb-1 weak points will reduce the expression of their respective
phenotypes in C. elegans.
Given that the stress reduction techniques are both of an immersive nature, the effectiveness of
the tests will not be diminished, as the entirety of the nematode is subjected to the experimental
effects.
Materials & Methods
Previous Examinations Nematode Strains
Figure 4: mua-6/ifa-3 Expression in Soft Agar
Retrieved from
http://wormatlas.psc.edu/hermaphrodite/musclesomatic/Im
ages/MusFIG11lr.jpg
Zhu, 2014
• IFA-2 and IFB-1 mirror human
myopathies
• Stress reduction
1. Soft Agar
2. unc-54
Heterochrony
Ambros, 1997
• Changes in timing of developmental
events
• lin-14 (gf/lf)
• mua-6
• ifa-3
• lin-14 (gf/lf)
• unc-54
Figure 3
Retrieved from http://ac.els-cdn.com/S0012160603004652/1-s2.0-S0012160603004652-
main.pdf?_tid=fe03ba12-9886-11e3-a92e-
00000aab0f26&acdnat=1392719310_5654fadfa6af33b70af2f2b6c9b6f6ca
Temporal vs Osmotic Regulation
• Soft Agar Experiments
• unc-54 Experiments
• Control against mua-6/ifa-3
• A, B, C categorization
lin-14 (gf)
lin-14 (lf)
Soft Agar Unc-54
mua-6 mua-6
mua-6/gf mua-6/gf
mua-6/lf mua-6/lf
ifa-3 ifa-3
ifa-3/gf ifa-3/gf
ifa-3/lf ifa-3/lf
+/+ ♂ x mua-6/mua-6 ♀♂
+/mua-6 ♂ x unc-54/unc-54 ♀♂
+/unc-54 ;+/mua-6
+/unc-54 ;+/+
No mua-6
progeny
+/? ;mua-6/mua-6
unc-54/unc-54 ;mua-6/mua-6
Stage 1
Stage 2
Stage 3
Stage 4
Stage 5
unc-54 Cross
X2=7.212 p=0.71
Figure 5: mua-6/ifa-3 Expression with unc-54
X2=6.508 p=0.77