INTRODUCTION It is known that neurodegenerative disorders such

as Alzheimer’s disease and Parkinson's disease are caused by protein accumulation within in the body (Takalo et al, 2013). Studying these disorders in human subjects can sometimes be difficult. Often, the use of a model organism like Caenorhabditis elegans can be used for preliminary studies. C. elegans has been shown to produce protein accumulation in response to a stress (Morton and Lamintina, 2013). Dr. Boyd’s Lab at Middle Tennessee State University have shown that ubiquitin and proteasome fluorescent tagged C. elegans respond to mechanical and osmotic stress by re-localizing ubiquitin and proteasome into spheres within the nucleus of the oocyte. It is not known if the re-localized ubiquitin and proteasome cause the oocyte of the adult to die. This poster aims to address two questions, the first question is “Do the spheres cause the oocyte death” and the second question is “Can the experimental worms survive and lay viable eggs”?

Ubiquitin-Proteome Spheres In Post Stress Response RecoveryHeather Corban, Central Magnet

Katherine Sampuda, Middle Tennessee State University

DISCUSSION

Both the ubiquitin and the proteosome strains of C. elegans had similar results. In both strains, death increased as the as time of exposure and molar concentration increased. Both strains also had spheres of relocalized protein after salt exposure. These data suggest that worm death and sphere formation may be correlated. However, we do not have any data yet that confirms a connection between sphere formation and worm death. In addition, we do not have data on the viablity of eggs in the surviving worms. More research is needed to address these two points.

ACKNOWLEDGEMENTSSpecial thanks to Middle Tennessee State University TRIAD GK12 program sponsored by a National Science Foundation Grant.

Thanks, also to Dr. Boyd for her expertise.

RESULTSIn both strains, death increases as the time exposed to the salt increases (Graph 1) as well as with an increase in molar concentration (Graphs 1 and 2). The ubiquitin and proteasome strain contain spheres of re-localized protein after each time point (Figure 1 and Figure 2)

Graph 1 Proteasome and Ubiquitin Survival at 2M NaCl

Graph 2 Proteasome and Ubiquitin Survival at 1M NaCl

Figure 1 Ubiquitin Tagged C.elegans At 25minutes

Figure 2 Proteasome Tagged C.elegans At 25minutes

MATERIALS & METHODS The experiment was duplicated using two

transgenic strains of C. elegans, a ubiquitin labeled strain and proteasome labeled strain. In both cases, 10 worms were placed in each of the following osmotic solutions: 1M NaCl, 2M NaCl, and 5M NaCl. The worms stayed in the salt solutions for one of three time frames: 10 minutes, 25 minutes, and 45 minutes. After that time, the worms were transferred to M9 Buffer solution and left for an equivalent number of minutes in order to wash the salt off the worm. The worms were observed on a confocal microscope to determine if any spheres were present after the experiment. The next day, survival numbers were recorded from the experimental worm plates.

REFERENCESLant B. and K.B. Storey. 2010. An Overview of Stress Response and Hypometabolic Strategies in Caenorhabditis elegans: Conserved and Contrasting Signals with the Mammalian System. Int J Biol Sci 6(1):9-50.

Morton E.A. and T. Lamintina. 2013. Caenorhabditis elegans HSF-1 is an essential nuclear protein that forms stress granule-like structures following heat shock. Aging Cell 12(1): 112-20.

Takalo M., A. Salminene, M. Hiltunen and A. Haapasalo. 2013. Protein aggregation and degradation mechanisms in neurodegenerative diseases. Am J Neurodegener Dis 2(1): 1-14.

10 25 450

2

4

6

8Proteasome Tagged C.elegan

Survival At 2M NaCl Solution

Time (minutes)

Num

ber O

f Wor

ms

That

Sur

vive

d10 25 45

02468

1012

Ubiquitin Tagged C.elegan Survival At 2M NaCl Solu-

tion

Time (minutes)

Num

ber O

f Wor

ms

That

Sur

vive

d

Protea-some

Tagged C.elegan Survival At 1M NaCl

Protea-some

Tagged C.ele-gan

Survival At 1M NaCl

Proteasome at 25 minutes

Proteasome at 10 minutes

Ubiquitin at 10 minutes

Ubiquitin at 25 minutes

10 25 450

1

2

3

4

5

6

7

8

Proteasome Tagged C.elegan Survival At 2M NaCl Solution

Time (minutes)

Num

ber O

f Wor

ms T

hat S

urvi

ved

10 25 450

2

4

6

8

10

12

Ubiquitin Tagged C.elegan Survival At 2M NaCl So-lution

Time (minutes)

Num

ber O

f Wor

ms T

hat S

urvi

ved

10 25 4502468

1012

Ubiquitin Tagged C.elegans Survival At 1M NaCl Solution

Time (minutes)

Num

ber O

f Wor

ms T

hat

Surv

ived

10 25 4502468

1012

Proteasome Tagged C.elegan Survival At 1M NaCl

Time (minutes)

Num

ber O

f Wor

ms T

hat

Surv

ived

Worm Strain

[High Salt]

Time (min)

Avg Spheres Per Gonad

Avg Spheres

Per Oocyte

LN130 2M 10 46/50+ 5

LN130 2M 25 45/50+ 4

LN130 2M 45 50/50+ 4

LN153 2M 10 41/50+ 2

LN153 2M 25 46/50+ 5

LN153 2M 45 50/50+ 5

Ubiquitin and proteasome re-localized in sphere formation in both of the transgenic strains at each recovery time point.