Neuronal

Dierences in C. elegans Worms from Dierent Parts of the World

Natalie Bulger, Eli Solomon, Alec Friswold

Works Cited
Dolgin, Felix, and CuXer. "Hakuna Nematoda." Nature.com. Nature Publishing Group, 12 Dec. 2007. Web. 29 July 2014.
Wol, J. Carleton College, Biology Department. Carleton Summer Science Ins9tute Handouts. 2014.
Marinelli, Janet. Plant. New York, NY: DK, 2005. Print.
Acknowledgments

Discussion-Chemotaxis

Acetone

N2
LKC34
KR314

par-2

L4440
RNAi

Figure 7. Varied Reac1ons to par-2 RNAi. The

control showed no change in number of
ospring from original worms. N2 worms
ospring were fewer than the control when
aected by par-2 RNAi, LKC34 and KR314
worms ospring increased.

We would like to thank Professor Jennifer Wol J-Wol, Anais Boyd, Georgia SchmiX, the Carleton Summer Science Ins9tute, and the thousands of C. elegans.

Octanol

Cineole
Terpineol
Strains and Scents

KR314

LKC34

N2

KR314

LKC34

N2

KR314

LKC34

-1

N2

-0.5

KR314

18
16
14
12
10
8
6
4
2
0

Trial 2

LKC34

Figure 6. LKC34 Exhibits Dierent unc-4 RNAi Reac1on. The control plates
showed no change in any worm strain. The unc-4 RNAi had no eect on
N2 or KR314, but LKC34 exhibited a 25% dierence in worms aected by
RNAi. These worms were not able to crawl backwards.

Trial 1

N2

Gene Reac1ons

KR314

The results of the unc-4 RNAi experiment demonstrated dierent

neuronal reac9ons among the three strains of C. elegans. unc-4 RNAi
inhibits C. elegans ability to crawl backwards by limi9ng gene expression
in certain neurons. When the control, L4440 RNAi, was fed to the three
strains it resulted in 100% backwards mobility in all three strains of
worms, as predicted. However, LKC34 (Madagascar) reacted dierently
than the others a]er being fed unc-4 RNAi. While 100% of the N2 (United
Kingdom) strain and KR314 (Canada) strain were able to move backwards
a]er being fed unc-4 RNAi, only 75% of the LKC34 strain were s9ll able to
move backwards a]er being fed the same unc-4 RNAi. The dierent
reac9ons of LKC34 to unc-4 RNAi demonstrate that this strain of C.
elegans does, in fact, have dierent neuronal reac9ons than the other
two strains. This supports the idea that C. elegans from dierent parts of
the world vary in neuronal func9on.

The par-2 RNAi experiment had very unexpected results. par-2 RNAi
aects the worms reproduc9ve tract and is supposed to inhibit C.
elegans ability to produce viable ospring. As predicted, more live adults
from the N2 strain survived a]er being fed the control RNAi, L4440, rather
than when fed the par-2 RNAi. However, both the LKC34 and the KR314
strain resulted in more live adults on par-2 RNAi, than on L4440 RNAi. In
fact, KR314 had three 9mes as many live adults survive on par-2 RNA,
than on L4440. The increase in worms a]er introducing par-2 RNAi is
curious, and should be inves9gated further. However, if these results are
reproducible, the experiment shows that the three dierent strains of
worms do exhibit extremely dierent reac9ons to par-2 RNAi.

LKC34

Discussion-RNAi

Results

N2

1. 4-6 adult hermaphrodites of N2, LKC34, and KR314 strains were fed on
agar plates seeded with bacteria containing double stranded RNA. One
set of plates was the control with L4440 RNAi, the second set
contained unc-4 RNAi, and the third set contained par-2 RNAi.
2. All plates were incubated at 22 degrees Celsius.
3. A]er one day these hermaphrodites were transported to fresh plates.
4. All plates were returned to 22 degrees Celsius for two days.
5. The eects of unc-4 RNAi were tested by tapping the head of 20
worms and recording whether or not they were able to crawl
backwards.
6. The eects of par-2 RNAi were tested by dividing the plates into four
quadrants and recording how many live adults were in quadrant one.
7. The eects of L4440 RNAi on backwards movement and the number of
live adults were also recorded as a control to compare to par-2 and
unc-4 RNAi.







Figure 4. Three RNAi example plates for one strain

Chemotaxis Index

RNAi Methods

Number of Live Adults

Caenorhabdi6s elegans is a free-living soil nematode, 1 mm in length, that

is commonly used for studying the nervous system. C. elegans worms are
rela9vely inexpensive and easy to maintain. They contain only 302
neurons, making their nervous system easily studied and understood.
Although simple, they bear striking similari9es to more complex
organisms, including humans, in both their nervous system and other
9ssue types. C. elegans are an ideal model organism for neuroscience
research.

Experiments
unc-4 RNAi
par-2 RNAi
Chemotaxis
The goal of the unc-4 RNAi
experiment was to nd potenn9al
dierences in reac9ons to
restric9ng gene expression in
neurons. The goal of the par-2
Figure 2. Chemosensory Neurons of an N2 individual. The neurons are
RNAi experiment was to show if
stained with Di-I. No dierences between the chemosensory neurons in
there are dierences in gene
N2, LKC34, and KR314 were observed.
expression throughout the rest of
Chemotaxis Methods
the body. The goal of the
To test the chemosensory neurons in the N2, LKC34, and KR314 strains of
chemotaxis experiment was to
C. elegans, a chemotaxis experiment tested the worms' reac9ons to
nd out if strains from the United
dierent odorants. Two trials of this experiment were completed.
Kingdom (N2), Madagascar
1. 15 agar plates were divided into sec9ons of aXrac9on, repulsion, and
(LKC34), and Canada (KR314)
neutral.
diered in their chemotaxis Figure 1. Odorant and Strain Loca1on. Pine trees are found in the same places as the N2 and KR314 strains
of C. elegans, Vanilla bean is found in the same place as LKC34, and Eucalyptus is not found with any strains 2. 1l of sodium azide was placed 5 mm from the edge of the plate.
behavior.

Sodium azide paralyzes the worms once they have moved closer or
farther from the odorant being tested.
3. Ethanol was placed on the repulsion side of the plate.
4. 3 l of the odorant being tested was diluted in ethanol and then
placed on the aXrac9on side of the agar plate.
5. One drop of C. elegans was placed directly in the center of the plate in
the neutral por9on.
Chemotaxis
RNAi
6. A]er the plates had sat for 2 hours, they were refrigerated overnight.
KR314 Social Clump
7. The worms that had seXled in each por9on of the plate were counted
The dierence in aXrac9on or repulsion of
The RNAi experiments introduced bacteria
LKC34 Social
the next day.
dierent strains to specic odorants can
that inhibited unc-4 and par-2 gene
Clump
8. A formula ((AXracted - Repelled)/total worms) was used to determine
demonstrate neuronal dierences in C.
expression to compare the worms
the chemotaxis index of each strain of worm with a specic odorant.
elegans nervous systems.
neuronal and reproduc9ve reac9ons.

100%

80%

N2
60%

LKC34
40%

1
KR314
20%

0%


Can move Cannot Can move Cannot

0.5
backward
move
backward
move

backward
backward

Figure 3. Example chemotaxis plate
unc-4
L4440

0
% Move Backwards

Introduc1on

Vanillin

Figure 5. Similar Chemotaxis Reac1ons. All strains

reacted similarly to the controls, Acetone and
Octanol. All strains were repelled by Terpineol. The
reac1ons to Cineole varied. N2 was repelled, KR314
was aWracted, and LKC34 was dierent in each trial.
The reac1ons to Vanillin also varied. LKC34 was
aWracted, KR314 was mostly aWracted, although in
the rst trial it was slightly repelled, and N2 was
dierent with each trial.

N2 Solitary Spread

The chemotaxis experiment showed similar trends for all three strains.
They all reacted to the control odorants, acetone and octanol, as
predicted. Their reac9ons to cineole, terpineol, and vanillin were also very
similar. Although they diered in aXrac9on and repulsion to cineole and
vanillin between the two trials, all of the strains displayed a very small
reac9on in terms of magnitude. Also, all three strains showed similar
repulsion reac9ons to terpineol. The similar reac9ons displayed by all
three strains demonstrate that although they originated geographically
isolated from one another, their chemosensory neurons do not dier
signicantly to these specic odorants.

Conclusion

We predicted that geographic dierences between N2, LKC34, and KR314

C. elegans would result in neuronal varia9on within their nervous
systems. The chemotaxis experiment did not support this idea because all
three strains showed similar trends in terms of their reac9ons to dierent
odorants. However, the unc-4 RNAi experiment did support this idea
because the LKC34 strain from Madagascar reacted dierently than N2
and KR314 when fed unc-4 RNAi. Likewise, the par-2 RNAi experiment
resulted in very dierent reac9ons from all three strains. Due to the
varied results from all three experiments, further study is needed on the
eect of geographic origin on the dierences of the nervous system of C.
elegans.