The Effect of Celecoxib on Alzheimers Disease Symptoms in

Drosophila melanogaster
Lily Shearer
Niles North High School
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Table of Contents

Acknowledgements..2

Purpose, Hypothesis, and Rationale.3

Review of Literature.....4

Materials..10

Procedure.11

Variables.13

Data.....15

Graphs.....19

Discussion............22

Statistical Analysis.......24

Error Analysis..26

Conclusion...27

References....28
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Acknowledgements

I could not have completed this project without the guidance of so many people.

First of all, I would like to thank my teacher and sponsor Mrs. Camel for all of the help

throughout the duration of my project. Thank you for always being so patient with me and for

giving such great comments on every aspect of my project. I would not be here if it was not for

you and your help.

I would also like to extend thanks to the other STEM teachers for all of their help with getting

supplies, providing support, and much more throughout my experimentation.

A special thanks to my classmates for listening as I went through all of my ideas and always

providing moral support when I needed it. I am so lucky to have classmates and friends who are

so helpful and so supportive.

Finally, I would like to thank my parents for always pushing me to do better and for all of their

support throughout all of my life.

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Purpose
The purpose of this experiment is to see if anti-inflammatory medication, specifically celecoxib,

can reduce Alzheimer's symptoms in Drosophila melanogaster by targeting the buildup of

amyloid plaques in the brain.

-Alzheimers disease is one of the most prevalent diseases in our nation, and it is important to

continue to target the specific causes of this disease and how we can use existing treatments to

help reduce harmful symptoms.

Hypothesis

If Drosophila melanogaster that have a mutant in the Amyloid Beta A4 Precursor Protein (APP)

gene and exhibit Alzheimer's symptoms are given celecoxib, an anti-inflammatory medication,

then their Alzheimer's symptoms will be less significant and their memories will function better

in a T-maze.

Rationale
APP is one of the three main genes that cause FAD Alzheimer's. These mutants cause the

buildup of amyloid plaques in the brain due to the A-42 peptides. Mefenamic acid has been

known to prevent the buildup and attenuating the neurotoxicities, and celecoxib has similar

chemical properties to mefenamic acid. If this buildup is prevented, the Alzheimers symptoms

will be less significant and the fruit flies will be able to function better.

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Review of Literature
Alzheimers disease (AD) is the most prevalent form of dementia in the human brain. In

2010, an estimated 36 million people were living with AD. This figure is predicted to increase to

66 million by 2030 (Newman, M., Ebrahimie, E., & Lardelli, M, 2014). The main feature of AD

is progressive memory loss, but symptoms can also include depression, delusions, hallucinations,

and aggressive behavior. Alzheimer's is a very prevalent disease in our society, and although

research is extensive, it is important to look into all possible treatments that have the possibility

of making a difference.

There are two main types of AD, which can classed as familial (FAD), usually which

onsets when the person is under 65 years of age, and sporadic (SAD), which onsets after the

person has turned 65. FAD is often attributed to mutations in three genes, PRESENILIN 1

(PSEN1), PRESENILIN 2(PSEN2), and the AMYLOID BETA A4 PRECURSOR PROTEIN (APP)

(Newman, M., Ebrahimie, E., & Lardelli, M, 2014). The PSEN proteins as well as the APP

proteins comprise -secretase complexes. These complexes are responsible for the cleavage of a

number of single-pass transmembrane proteins such as APP and NOTCH. APP is initially

cleaved by - or -secretases to release APP or APP fragments, respectively. The subsequent

cleavage of APP by -secretase after -secretase cleavage liberates amyloid- (A) peptides of

various lengths. The longer A-42 peptide is prone to aggregation and is suggested to form toxic

oligomers and fibrils that eventually deposit as amyloid plaques in the brain (Newman, M.,

Ebrahimie, E., & Lardelli, M, 2014). Amyloid plaques consist of a proteinaceous core composed

of 510nm amyloid fibrils surrounded by dystrophic neurites, astrocytic processes and

microglial cells. These amyloid plaques triggers subsequent cellular abnormalities such as

inflammation and oxidative stress, as well as an unbalance of neurotransmitters in the brain.

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These ultimately lead to neuronal dysfunction, degeneration and death, and have been

hypothesized to be one of the lead causes of FAD Alzheimer's. (Newman, M., Ebrahimie, E., &

Lardelli, M, 2014)

Figure 1: (Imbimbo, B. P., Solfrizzi, V., & Panza, F, 2010).

There are 4 drugs currently approved by the FDA to treat AD, and can be classified into

two groups: Cholinesterase inhibitors and memantine (Alzheimers Association, 2016).

Cholinesterase inhibitors inhibit enzymes from breaking down acetylcholine. Acetylcholine is an

important neurotransmitter for memory, and because it is in short supply in people with AD,

delaying this destruction creates a greater chance of it being passed on to the next nerve cell

(Alzheimer's Association, 2016). However, cholinesterase inhibitors usually only delay the onset

of Alzheimers symptoms. Donepezil, galantamine and rivastigmine are all types of

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cholinesterase inhibitors (Alzheimer's Association, 2016). Memantine, the fifth Alzheimer's drug,

is an NMDA (N-methyl-D-aspartate) receptor antagonist, and it works by regulating glutamate,

another important neurotransmitter in the brain involved in learning and memory. The

attachment of glutamate "docking sites" on the cells surface called NMDA receptors allows

calcium to enter the cell. This process is important for cell signaling, as well as learning and

memory (Alzheimers Association, 2016). However, in patients with AD, excess glutamate is

sometimes released from damaged cells, which leads to calcium overexposure and can speed up

cell damage. Memantine assists the prevention of this overexposure by partially blocking the

NMDA receptors (Alzheimer's Association, 2016). The success of these medications varies from

patient to patient, but overall usually only delay the symptoms for 2-5 years (Alzheimer's

Association, 2016).

Drosophila melanogaster, or fruit flies, have become commonly used as a model

organism for genetics, microbial pathogenesis, physiology, life history evolution, and more, due

to their short lifespan(40-60 days) and similarities to the human genome (Pitman, J. L., 2009).

Because of the wide knowledge of the fruit fly genome, it has become possible to breed fruit flies

that have mutations in specific genes, including APP (Pitman, J. L., 2009). One of the main

reasons behind the use of fruit flies in Alzheimers research is that the fly has a brain of

approximately 200,000 neurons, and like the vertebrate central nervous system, it is composed of

a series of functionally specialized substructures. The mushroom bodies deal with memory

consolidation and are analogous to the human hippocampus (Moloney, A., Sattelle, D. B.,

Lomas, D. A., & Crowther, D. C, 2010). The neurons of fruit flies are also very similar to those

of humans in terms of their shape, synaptic intercommunications and their biochemical

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signatures (Moloney, A., Sattelle, D. B., Lomas, D. A., & Crowther, D. C, 2010). These

functional and structural similarities allow fly models of human disease to be successful on the

biophysical, molecular biological, neurobiological and behavioural levels (Moloney, A., Sattelle,

D. B., Lomas, D. A., & Crowther, D. C, 2010). In a study done in 1990, it was concluded that

flies retrieve memory efficiently when rewarded sugar, and that sugar reinforced memory

persists much longer than shock reinforced memory.

One test on NSAIDs and Alzheimers symptoms was done in 1993 that produced positive

results A first 6-month, double-blind, placebo-controlled study in 44 AD patients showed a

significant slower cognitive decline in patients receiving indomethacin compared to those on

placebo (Rogers et al., 1993). Unfortunately, 42% of patients abandoned the study due to adverse

events. Thus, the positive results of this study are of difficult interpretation. A subsequent 1-year

double-blind, placebo-controlled, study of indomethacin (100mg/day) given with omeprazole

(20mg/day) as gastroprotective agent, to 51 mild-to-moderate AD patients produced relatively

low drop-out rates (7 out of 26 patients on indomethacin and 6 out of 25 patients on placebo).

Unfortunately, the low number of subjects rendered the study inconclusive with non-significant

trends in favor of indomethacin on two cognitive scales at 12months (de Jong et al., 2008).

In a recent study, mice were used as models to test if mefenamic acid could reduce

Alzheimers symptoms. Mefenamic acid is an non-steroidal anti-inflammatory medication

(NSAID) that is a cyclooxygenase-1 and 2 inhibitor with only moderate anti-inflammatory

properties (Daniels, M. J., Rivers-Auty, J., Schilling, T., & Spencer, N. G, 2016). After mice that

were genetically modified to have Alzheimer's symptoms were injected with mefenamic acid,

their symptoms were almost completely gone. It was concluded that mefenamic acid attenuates
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the neurotoxicities induced by the A-42 peptide, or the amyloid plaques, and specifically targets

the NLRP3 inflammasome (Daniels, M. J., Rivers-Auty, J., Schilling, T., & Spencer, N. G,

2016).

Mefenamic acid has been identified as a NLRP3-selective inhibitor. Fenamate NSAIDs

have been shown to inhibit IL-1 secretion from macrophages, although the significance of COX

inhibition remains unclear; The fenamate class of NSAIDs inhibit the NLRP3 inflammasome via

reversible blockade of volume-regulated anion channels (VRAC) in the plasma membrane (Dean

L, 2011). Celecoxib is a part of the fenamate class of NSAID. Because of this, it can be

concluded that celecoxib may have enough similar properties to mefenamic acid that would

allow it to also reduce amyloid plaques and/or the negative effects that these plaques cause.

Celecoxib is a nonsteroidal anti-inflammatory drug (NSAID) used to treat mild to moderate pain

(Dean L, 2011).

Celecoxib is a COX-2 selective nonsteroidal anti-inflammatory drug (NSAID). It is used

to treat the pain and inflammation of osteoarthritis, rheumatoid arthritis, ankylosing spondylitis,

acute pain in adults, painful menstruation, and juvenile rheumatoid arthritis (Dean L, 2011).

Celecoxib is compound 22; the 4-sulfamoylphenyl on the 1-pyrazol substituent is required for

COX-2 inhibition and the 4-methyl on the 5-pyrazol system has low hindrance to maximize

potency, while the 3-trifluoromethyl group provides selectivity (Dean L, 2011).

In conclusion, the prevalence of Alzheimer's disease causes it to be a significant problem

in our society that researchers should continue to study to look for a cure. Due to the orthologous

genes and ease of experimentation, Drosophila melanogaster is a model organism to perform

Alzheimer's research on. The drug, mefenamic acid, has properties that allow it to target the
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NPL3 inflammasome and possibly inhibit the buildup of amyloid plaques that cause Alzheimer's,

and due to the similarities of the drug celecoxib to mefenamic acid, celecoxib has the potential to

also prevent AD. Because of the potential of these drugs, experimentation should be completed

to see their power and get closer to a cure for Alzheimers.

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Materials
Drosophila melanoxgaster (wild type- not genetic modification) (25-30 specimen)

APP Genetically Modified Drosophila melanogaster (75-95 specimen)

Celecoxib

Formula 4-24 Drosophila Medium, Plain (1 Liter)

Freezer

Drosophila Glass Culture Tubes (10-30)

Cylinder Sponges (30)

T Maze Materials (3 tubes connected to two culture vials)

Liquid odor 1 (resembles licorice)

Liquid odor 2 (resembles sweaty socks)

Sucrose

Balance

Tap Water

10 mL Beakers (2-4)

Disposable Pipettes

Stirring Rod

Scoopula

Graduated Cylinder

Ruler

Masking Tape

Gloves and Face Mask

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Procedure

Preparing Habitats
1. Obtain Formula 4-24 Drosophila Plain Medium and twelve culture tubes. Add
approximately 4 grams of dry media and approximately 10 mL of water to the bottom of
each culture tube.
2. Make sure to add enough water to the media flakes until an accurate solid consistency is
reached. The six culture tubes should be filled of the way with the media food source.
3. Using a mortar and pestle, break the celecoxib pills apart until they reach a fine powder.
4. Add 0.15 grams of celecoxib to three of the culture tubes. Add .30 grams of celecoxib to
another three of the culture tubes. Make sure the tubes are labeled with low
concentration and high concentration.
5. Place approximately 25 genetically normal flies in three of the vials. Label these
control.
6. Place approximately 25 APP modified fruit flies in each of the tubes that were given
celecoxib as well as 25 APP modified flies in the remaining three tubes.
7. Place all twelve labeled tubes in a stable, dry area. Make sure to plug the tops of the tubes
with foam sponges to prevent escape.
8. After one week, if necessary, transfer flies into new vials with fresh medium; render the
flies unconscious by placing each tube on a horizontal slant inside a freezer for
approximately 1-2 minutes. Take the tubes out and lay them down on their side. While
the fruit flies are unconscious, transfer all the specimen into their newly assigned tube.
Transfer the fruit flies by holding the frozen tube at a slant and slowly pouring the
unconscious specimen into the newly prepared tube also held at a slant. After the transfer
is complete, immediately cap the new tubes to prevent escape and place them on a
horizontal slant.

Training Flies
1. After a few days, begin to train the fruit flies to recognize the odor.
2. Place four drops of odor one on a foam sponge. Place in the vial and leave for 2 minutes.
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3. Immediately after, place another sponge and 1.25 grams of sugar in vial. Leave for 5
minutes.
4. Repeat step two for all tubes for two to three days.
5. Place four drops of odor two on a foam sponge. Place in the vial and leave for 3 minutes.
Immediately after, place a fresh, new sponge in vial.

T Maze Testing
1. Connect three plastic tubes together in a T formation according to box, with the ends of
the T having two additional tubes that have the sponge toppers. Place a sponge topper
with four drops odor 1 on the left side and a sponge topper with four drops of odor 2 on
the right side.
2. Anesthetize flies (according to step 8 above) and then place in the edge of the T maze.
Close the edge with a foam stopper.
3. Once flies have picked a side, remove the left tube, place 1.25 grams of sugar in, and
leave for 5 minutes. Anesthetize flies and return to previous tube.
4. Remove right tube, anesthetize flies, and return to previous tube.
5. Repeat testing for each vial for 4 days.

Disposal
1. Once testing is complete, euthanize flies by placing in the freezer for 15 minutes. Place
flies in a plastic bag and throw in garbage.
2. Dispose of the old food medium sources by rinsing the culture tubes with water and soap.
3. Wash out the tubes and any equipment used to touch the fruit fly specimen during the
duration of the experiment.
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Variables

Independent Variable: The independent variables in this experiment are the amount of
celecoxib that the fruit flies are exposed to. The amounts of 0.15 grams and 0.30 grams were
used for this experiment. The differing amounts of celecoxib have the possibility to change the
memory functioning of the fruit flies, and therefore are the independent variables.

Dependent Variable: The dependent variable is the number of fruit flies that choose odor 1,
which is the odor that had previously led them to sugar. This will be translated into percentages.
This is the dependent variable because the amount of flies that choose the first odor have the
possibility to change if they are given the celecoxib. The memory functioning of the fruit flies is
the dependent variable because it can possibly be changed by the amount of celecoxib that it is
exposed to.

Control Groups: The control groups are the group of flies that do not have a genetic mutation as
well as the group of fruit flies that have the mutation but are not given the celecoxib. These are
the controls because they demonstrates the response of no fruit flies to the drugs as well as how
fruit flies that have a mutation in APP perform in the T Maze. These are the controls because
they will not be affected by the celecoxib.

Constants:
- Environment of flies: food given each day, water, space in vial, temperature, light source,
etc.
- Amount of time flies are anesthetized for
- Materials used to gather results
These things were kept constant to prevent experimental error. By keeping the only variable that
changes the amount of celecoxib they were exposed to, the results would be more focused on
how the drug affects the memories of the fruit flies, instead of how these other variables affect
the memory functioning.
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Data

APP Mutation with Low Concentration of Celecoxib (.15g)

Odor 1 (led to Total Fruit Percent that

Vial 1 sucrose) Odor 2 Flies chose odor 1
Day 1 17 13 30 56.66666667
Day 2 14 14 28 50
Day 3 16 12 28 57.14285714
Day 4 16 11 27 59.25925926

Odor 1(led to Total Fruit Percent that

Vial 2 sucrose) Odor 2 Flies chose odor 1
Day 1 16 15 31 51.61290323
Day 2 17 15 32 53.125
Day 3 15 16 31 48.38709677
Day 4 18 15 33 54.54545455

Odor 1(led to Total Fruit Percent that

Vial 3 sucrose) Odor 2 Flies chose odor 1
Day 1 16 15 31 51.61290323
Day 2 15 16 31 48.38709677
Day 3 17 15 32 53.125
Day 4 17 13 30 56.66666667
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Averages for Low Concentration

Low
Concentration Average
of Drug Percent
Day 1 53.29749104
Day 2 50.50403226
Day 3 52.88498464
Day 4 56.82379349

APP Mutation with High Concentration of Celecoxib (.30g)

Odor 1(led to Total Fruit Percent that

Vial 1 sucrose) Odor 2 Flies chose odor 1
Day 1 17 14 31 54.83870968
Day 2 19 12 31 61.29032258
Day 3 19 16 35 54.28571429
Day 4 21 14 35 60

Odor 1(led to Total Fruit Percent that

Vial 2 sucrose) Odor 2 Flies chose odor 1
Day 1 17 14 31 54.83870968
Day 2 19 13 32 59.375
Day 3 15 16 31 48.38709677
Day 4 21 13 34 61.76470588
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Odor 1(led to Total Fruit Percent that

Vial 3 sucrose) Odor 2 Flies chose odor 1
Day 1 20 14 34 58.82352941
Day 2 20 13 33 60.60606061
Day 3 21 14 35 60
Day 4 21 13 34 61.76470588

Averages for High Concentration

High Average
Concentration Percent
Day 1 56.16698292
Day 2 60.4237944
Day 3 54.22427035
Day 4 61.17647059

APP Mutation with NO Celecoxib

Vial 1 (no Total Fruit

drug) Odor 1 Odor 2 Flies Percent
Day 1 16 14 30 53.33333333
Day 2 15 16 31 48.38709677
Day 3 16 15 31 51.61290323
Day 4 14 17 31 45.16129032
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Vial 2 (no Total Fruit

drug) Odor 1 Odor 2 Flies Percent
Day 1 16 15 31 51.61290323
Day 2 17 16 33 51.51515152
Day 3 14 18 32 43.75
Day 4 15 16 31 48.38709677

Vial 3 (no Total Fruit

drug) Odor 1 Odor 2 Flies Percent
Day 1 16 14 30 53.33333333
Day 2 17 15 32 53.125
Day 3 15 18 33 45.45454545
Day 4 16 14 30 53.33333333

Averages for APP Mutation without Celecoxib

Average
No Drug Percent
Day 1 52.75985663
Day 2 51.00908276
Day 3 46.93914956
Day 4 48.96057348

Control (No Mutation)

Vial 1 Total Fruit

(control) Odor 1 Odor 2 Flies Percent
Day 1 23 13 36 63.88888889
Day 2 23 14 37 62.16216216
Day 3 24 15 39 61.53846154
Day 4 24 14 38 63.15789474
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Vial 2 Total Fruit

(control) Odor 1 Odor 2 Flies Percent
Day 1 21 14 36 58.33333333
Day 2 20 14 34 58.82352941
Day 3 21 13 34 61.76470588
Day 4 22 14 36 61.11111111

Vial 3 Total Fruit

(control) Odor 1 Odor 2 Flies Percent
Day 1 21 13 34 61.76470588
Day 2 22 14 36 61.11111111
Day 3 23 13 36 63.88888889
Day 4 24 12 36 66.66666667

Averages for Control

Average
Control Percent
Day 1 61.32897603
Day 2 60.69893423
Day 3 62.3973521
Day 4 63.64522417
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Graphs
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Slopes of Best Fit Lines

Control(no mutation): 0.87

Low Concentration: 1.29
High Concentration: 0.88
APP with no Celecoxib: -1.56
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Discussion

When looking at this data, it is clear that on a day to day basis and on average, the fruit

flies that had the APP mutation and were exposed to the celecoxib chose the positive odor(the

one that led to sucrose) more frequently than the flies with the APP mutation who were not

exposed to the celecoxib. The fruit flies that had the genetic mutation but were not given

celecoxib typically chose the positive odor 45-55 percent of the time, while the fruit flies given

0.15 grams of celecoxib chose the positive odor around 50-55 percent of the time and the fruit

flies given 0.30 grams of celecoxib chose the positive odor around 55-60 percent of the time. The

control group (the flies with no genetic mutation) chose the positive odor 60-65 percent of the

time. It is obvious that the group given a higher concentration of celecoxib had a greater

improved memory ability than the group with given a lower concentration, meaning that greater

amounts of celecoxib did cause a greater improvement in the memory functioning in the fruit

flies. This demonstrates that the celecoxib did help reverse some of the Alzheimers symptoms

that were present in the flies with the mutation because the fruit flies were choosing the odor that

they remembered having sugar, a positive result, with that odor. However, both groups were still

significantly lower than the control group, showing that the drug did not completely repair the

damaged memory functioning.

It is also possible to see if the memories of the fruit flies continued to progress each day

when looking at the slope of the best fit line on the scatter graph. When the slope is positive, it

means that more and more flies are choosing the positive odor each test. This can be attributed to

the fact that they are continually being trained by receiving the sucrose, and so they will continue

to pick the sucrose and more flies will choose the left side, but it can also be attributed to the fact
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that their memories are continuing to improve because of the drug. Because the slope of the APP

mutation control group is negative, it can be concluded that the fruit flies memory is continuing

to deteriorate each day. The groups given the drug as well as the control group improved on

average each day (given the positive slope), which demonstrates that the celecoxib is making a

difference in the memory ability of the fruit fly because it is not behaving the same as the group

with no drug, and it is also showing that the fruit flies were able to register the positive results

and continue to choose the odor that led to the sucrose.

The results of the APP mutated flies fluctuated up and down more than the results of the

control group as well as the groups that were given the celecoxib. This demonstrates a greater

severity of Alzheimers symptoms in the fruit flies because they are behaving and choosing

which odor more sporadically. Because the groups given the celecoxib did not fluctuate as much,
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Statistical Analysis

0.10 grams Celecoxib

Compared to APP with no Celecoxib

Null hypothesis: The average percentage of fruit flies given no celecoxib that chose the odor
with a positive result is equal to the average percentage of fruit flies that were given celecoxib
and chose the odor with a positive result.

Alternative Hypothesis: The average percentage of fruit flies given no celecoxib that chose the
odor with a positive result is equal to the average percentage of fruit flies that were given
celecoxib and chose the odor with a positive result.

Standard Deviation of Experimental Group: 3.559178019

Standard Deviation of Control Group: 14.25630647
Sample Size: 12
Degrees of Freedom: 11
t= 2.38361
p=.03627

Because p<.05, this data is statistically significant and the null hypothesis is rejected in favor of
the alternative hypothesis.

0.30 grams Celecoxib

Compared to APP Mutation with no Celecoxib
Null hypothesis: The average percentage of fruit flies given no celecoxib that chose the odor
with a positive result is equal to the average percentage of fruit flies that were given celecoxib
and chose the odor with a positive result.

Alternative Hypothesis: The average percentage of fruit flies given no celecoxib that chose the
odor with a positive result is equal to the average percentage of fruit flies that were given
celecoxib and chose the odor with a positive result.

Standard Deviation of Experimental Group: 4.075691875

Standard Deviation of Control Group: 14.25630647
Sample Size: 12
Degrees of Freedom: 11
t=5.177818
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p=.000305
Because p<.05, this data is statistically significant and the null hypothesis is rejected in favor of
the alternative hypothesis.

When performing statistical analysis, the results of the experimental groups were

compared with the results of the group with the APP mutation, but that were not given the

medication. For the group with the low concentration of celecoxib, the t value was calculated to

be 2.38361, and the p was calculated to be 0.03627. For the group with the higher concentration

of celecoxib, the t value was calculated to be 5.177818, and the p was calculated to be .000305.

Because the p value for both groups is less than 0.05, the null hypothesis is rejected in favor of

the alternative hypothesis, which means that the data is statistically significant.
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Error Analysis

There were many opportunities for experimental error when conducting this experiment.

One major example of experimental error is the fact that by working with living organisms, each

one is not identical and therefore has differences in their body structure and nervous system. I

attempted to rule out this error by using as many fruit flies as possible, but more fruit flies and

more data for my next attempt would be a good way to try and limit experimental error.

There is also the fact that because of the relatively short lifespan of fruit flies, the number

of fruit flies fluctuated on a day to day basis because some of them died at the same time that

many of them were reproducing. This creates a source of error because the fruit flies that were

being produced were not trained in the same way that the original flies were. This makes it so

that they will have more sporadic results. This was also attempted to be corrected by doing the

tests on a day to day basis so fewer of them had the chance to die or reproduce. The number of

flies also slightly fluctuated because a few of them would not move when put in the T-maze, and

this error was attempted to be corrected by tapping on the plastic in order to try and coax the fly

into choosing a side. This was not always successful, and in the future, I will attempt to build a

maze where the flies are limited in space and must choose a side.

Another possibility for experimental error is human error; even though exact

measurements were attempted for each step, there is always the possibility that something was

not measured correctly or was inserted incorrectly.I attempted to make as precise measurements

as possible to try and prevent this experimental error. To really combat the possibility of error,

more tests should always be completed.

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Conclusion

My hypothesis was supported by these results. These results showed that fruit flies with

an APP mutation had a more sporadic pattern when choosing the odor, while flies that had this

mutation but were given celecoxib tended to choose the odor that related to sucrose, which was

more similar to the control (the group with no mutation). This demonstrates that the celecoxib

caused the fruit flies to regain memory function, most likely by targeting the NLRP3

inflammasome and preventing the buildup of amyloid plaques. This data was also proven

statistically significant, showing that there is a real correlation between celecoxib and memory

functioning. While these findings did show that celecoxib did reduce AD symptoms, there is still

quite a bit of experimentation that should be done on how NSAIDs affect Alzheimers disease.

This project was on a much smaller scale, and while there are many similarities to

genetics and memory functioning of fruit flies and humans, there is still a long way to go before

this will be carried into an effective treatment for humans. Because Alzheimers is such a

devastating disease and has become so prevalent in our world, it is important to continue to look

at ways to treat it, and it is also important to look at existing medication and treatments to see

how they can be used to find a cure for this disease.

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References

Daniels, M. J., Rivers-Auty, J., Schilling, T., & Spencer, N. G. (2016, August 11). Fenamate
NSAIDs inhibit the NLRP3 inflammasome and protect against Alzheimers disease in
rodent models. In Nature Communications. Retrieved September 8, 2016, from
http://www.nature.com/ncomms/2016/160811/ncomms12504/full/ncomms12504.html

De Jong, D., Jansen, R., Hoefnagels, W., Jellesma-Eggenkamp, M., Verbeek, M., Borm, G., &
Kremer, B.
(2008). No Effect of One-Year Treatment with Indomethacin on Alzheimers Disease
Progression: A Randomized Controlled Trial. PLoS ONE, 3(1), e1475.
http://doi.org/10.1371/journal.pone.0001475

Dean L. (2011) Comparing NSAIDs. In: Dean L. PubMed Clinical Q&A [Internet]. Bethesda
(MD): National Center for Biotechnology Information (US); 2008-2013. Available from:
http://www.ncbi.nlm.nih.gov/books/NBK45590/

Imbimbo, B. P., Solfrizzi, V., & Panza, F. (2010). Are NSAIDs Useful to Treat Alzheimers
Disease or Mild
Cognitive Impairment? Frontiers in Aging Neuroscience, 2, 19.
http://doi.org/10.3389/fnagi.2010.00019

Levin ED, Cerutti DT. (2009) Behavioral Neuroscience of Zebrafish. In: Buccafusco JJ, editor.
Methods of Behavior Analysis in Neuroscience. 2nd edition. Boca Raton (FL): CRC
Press/Taylor & Francis; 2009. Chapter 15. Available from:
http://www.ncbi.nlm.nih.gov/books/NBK5216/
Meinertzhagen I. A., Hanson T. E. (1993). The development of the optic lobe, in The
Development of Drosophila Melanogaster, ed. Martinez-Arias M. B. A. A., editor.
(New York, NY: Cold Spring Harbor Press; ), 13631491.

Moloney A., Sattelle D. B., Lomas D. A., Crowther D. C. (2010). Alzheimers disease:
insights
from Drosophila melanogaster models. Trends Biochem. Sci. 35, 228235.
10.1016/j.tibs.2009.11.004

Newman, M., Ebrahimie, E., & Lardelli, M. (2014). Using the zebrafish model for Alzheimers
disease research. Frontiers in Genetics, 5, 189. http://doi.org/10.3389/fgene.2014.00189

Pitman, J. L., DasGupta, S., Krashes, M. J., Leung, B., Perrat, P. N., & Waddell, S. (2009). There
are many ways to train a fly. Fly, 3(1), 3.
 Shearer 29

Rogers J., Kirby L. C., Hempelman S. R., Berry D. L., Mcgeer P. L., Kaszniak A. W., Zalinski
J.,
Cofield M., Mansukhani L., Willson P., Kogan F. (1993). Clinical trial of indomethacin
in Alzheimer's disease. Neurology 43, 16091611

Poeck B., Strauss R., Kretzschmar D. (2012). Analysis of amyloid precursor protein function
in Drosophila melanogaster. Exp. Brain Res. 217, 413421.
10.1007/s00221-011-2860-3

Tang B. L., Liou Y. C. (2007). Novel modulators of amyloid-beta precursor protein

processing.

J. Neurochem. 100, 314323. 10.1111/j.1471-4159.2006.04215.x