Diet Responsive Transcription Factors That Influence Neurodegeneration in

Models of Alzheimer’s
𝟏,𝟐 𝟏 𝟏 𝟏 𝟏
Holly Jossart , Apurva Shah , Sudipta Bar , Pinky Kain , Pankaj Kapahi
𝟏 Buck Institute for Research on Aging, Novato CA, 94945; 𝟐 San Marin Highschool, Novato, CA, 94947

Introduction Big Question Lifespan analysis Activity Monitor

Will the knockdown of AL enriched
Alzheimer’s disease (AD) is an irreversible age-related disease that Females
destroys cognitive abilities and leads to dementia. AD causes transcription factors rescue the AL Females
widespread neuron damage (brain cell death). The neuron damage may
be caused by Beta-Amyloid and the Tau protein.
neurodegeneration caused by AD? 100 0.14

Healthy diet’s relationship to AD 80 0.12

Average bean crosses per fly

The consumption of certain foods is associated with the buildup of Tau

Methods

Survival (%)
and Beta-Amyloid. A poor diet increases the chances of developing AD 0.1
speeds up neurodegeneration. 60
0.08
Diet related Transcription Factors (TFs) 40
TFs are proteins possessing domains that bind to the DNA of enhancer Fig. 3 UAS-GAL4 System:
0.06
regions of specific genes. A large number of TFs have been found to be
diet related and are altered because of diet. The altering of these TFs • To express the TFs in AD flies. 20
related to diet could result in improvement in AD patients. The UAS-GAL4 system works 0.04
to activate gene expression.
The system has two parts: 0 0.02
the Gal4 gene, encoding 0 5 10 15 20
the yeast transcription 0
Elav Gal4/+ Elav Gal4>Tau Elav Gal4>Tau Elav Gal4>Tau Elav Gal4>Tau
activator protein Gal4, and the Days R406W R406W/ETS97D R406W/MYB R406W/GFZF
UAS (Upstream Activation RNAi RNAi RNAi
Sequence), an enhancer to AL Elav Gal4/+
AL Elav Gal4>Tau R406W DAY NIGHT
which GAL4 specifically binds
AL Elav Gal4>Tau R406W/ETS97D RNAi
to activate gene transcription. AL Elav Gal4>Tau R406W/MYB RNAi Fig. 7 Activity Monitor:
AL Elav Gal4>Tau R406W/GFZF RNAi For the activity monitor test, we place the flies in a system that
measures their activity levels during the day and night. AD flies show
Fig. 5 Lifespan analysis: more activity while normal flies have lower activity levels. All the TFs
Lifespan is kept track of to see how long the flies live. AD patients live a show rescue.
Fig. 1 Healthy Brain compared to AD brain: The normal brain (left) is Genetic Cross to generate rescue flies
shorter life than healthy people, as is with the flies. The Ad flies (red
• Elav GAL4/+
larger and healthier. The AD brain (right) is shrunken and unhealthy
because of the neuron damage. • Elav GAL4> UAS-TauR406W (AD Flies)
• Elav GAL4>UAS-Tau R406W/UAS RNAi EYS97D (Rescue)
line) die way faster than the normal flies (green line). All of the TFs
show rescue (black lines) because they die slower than the AD flies. Conclusion
• Each transcription factor knockdown showed a rescue in the AD
D. Melanogaster as our AD model • Elav GAL4> UAS-Tau R406W/UAS RNAi MYB (Rescue)
flies.
Humans share nearly 60% of genes with fruit flies. Flies have a fast • Elav GAL4>UAS-Tau R406W /UAS RNAi GFZF (Rescue)
• Partial rescue for eye phenotype, with all TFs
generation time and a short lifespan. Drosophila is an ideal disease
• For lifespan analysis, rescue is seen with all TFs
model system to study neurodegeneration.
How we tracked neurodegeneration Climbing •
•
For climbing no rescue is seen with any of the TFs
For Activity monitor, almost completely rescue with all TFs
Different diets - AL and DR Eye neurodegeneration Lifespan

Climbing Index Average (%)

Ad Libitum (AL- 5% yeast) and Dietary Restriction (DR- 0.5% Yeast) are Climbing analysis Activity Monitor 50
the main two diets used to understand the effect of diet on life span and
AD pathology. AL is considered a nutritious food and DR is restrictive.
Flies of all genotypes do better on DR. 40 Future Plans
ChIP Sequence
Using a public database and lab database, TFs chosen were highly
Results 30
• Investigate if putting crosses directly on AL and DR during
development stage can rescue the eye phenotype more
• Test with male flies
enriched on AL food. Rescue of Eye Neurodegeneration • Test the TFs together to see if rescue is increased
20
AL Based Transcription Factors
• ETS97D
• MYB
• GFZF
Fig 4 Rescue of Eye
Neurodegeneration:
Wild type eye (top left),
10 Acknowledgments
AD fly eye showing I would like to thank Claire Spafford for providing me the opportunity to
0 participate in the Buck Institute Summer Scholars Internship Program. I
How we modify flies based on TFs roughness and is smaller Elav Gal4/+ Elav Gal4>Tau Elav Gal4>Tau Elav Gal4>Tau Elav Gal4>Tau
Knockdown of TFs in AD flies using RNAi lines to determine if there is (top right), and other 3 R406W R406W/ETS97D R406W/MYB R406W/GFZF would also like to acknowledge the Buck Institute’s generosity for
an improvement of neurodegeneration in AD flies. showing rescue of the RNAi RNAi RNAi providing the resources needed for my experiments. Specifically, I
RNAi lines help to inhibit gene expression. neurodegeneration AD fly Genotypes would like to thank Professor Kapahi, Dr. Kain, and Dr. Bar for their
caused by the guidance throughout the summer.
Elav>TauR406w knockdown of each TF (3
120 Fig. 6 Climbing Index Average:
100
DR AL bottom)
To test climbing, we place flies in tubes without food. We make all the Bibliography
Survival (%)

80 Fig. 2 AD flies live flies go to the bottom by smacking the tubes on a table and then count
1. Greenspan, Ralph J. Fly Pushing: the Theory and Practice of Drosophila Genetics.
better and longer on how many cross a 6cm line in 10 seconds. We then take the percent Cold Spring Harbor Laboratory Press, 2004.
60
DR food compared to crossed and analysis the data. None of the TFs were able to rescue 2. Hiller, Abigail J., and Makoto Ishii. “Disorders of Body Weight, Sleep and Circadian
40 Rhythm as Manifestations of Hypothalamic Dysfunction in Alzheimer’s
AL food the climbing phenotype.
Disease.” Frontiers in Cellular Neuroscience, vol. 12, 2018,
20 doi:10.3389/fncel.2018.00471.
0 3. Tabuchi, Masashi, et al. “Sleep Interacts with Aβ to Modulate Intrinsic Neuronal
0 5 10 Excitability.” Current Biology, vol. 25, no. 6, 2015, pp. 702–712.,
Days 15 20 25
doi:10.1016/j.cub.2015.01.016.

www.postersession.com
www.postersession.com