Final Project

Hello, everyone.

I'm Zoe Vanderburgh, and today we're gonna be talking about

metabolites and their effect on brain behavior.

Metabolites are very useful molecules that control the activity of

many complex physiology.

However, their role in complex behaviors is far less understood,

and we will be looking more in depth that their

potential involvement in these behaviors.

The paper I looked into his on the rapid metabolic

shifts that arise from the transition between the hunger and

Fed states endorse a field, Lancaster or fruit flies from

mapping out these shifts.

Steps can start to be made in the process of

comprehending what role in metabolic signaling plays and complex behaviors,

such as feeding the goal of the papers to turn

away from the more acknowledged side of metabolites.

Their energetic value by shifting to less understood side of

their informational value.

Gaining knowledge on how these shifts and metabolites fit within

different cellular pathways is a challenge.

The studies done within the paper will be the perfect

ground working beginning the process.

I chose this paper as I am very interested in

learning more about how diets can change the gene regulation

patterns within the brain as well as the overall behavior

of the organism.

This is the figure that I'll be discussing further, but

just to give a rough overview of it.

For now, the figure is split into five subsections.

A few e sections A and B, are referring to

the metabolites that are changing within the bodies of fasted

and re fed fruit flies, while C and D referred

to the metabolite shifts in the head.

Lastly, Section E is depicting the overlap between the metabolic

shifts within the body and heads of these fasted and

re fed flies.

Now that we have the general gist of the picture,

let's get into the experimental methods used to gather the

data for it.

First, we have ultra high performance liquid chromatography tandem mass

spectroscopy or U P L C M S M.

S. For short.

It is powerful technique used for many applications, which has

very high sensitivity and selectivity.

Generally, it's application is oriented towards the general detection and

potential identification of chemicals in the presence of other chemicals,

and a complex mixture has both qualitative and quantitative uses.

For example, identifying unknown compounds, determining the isotopic composition of

elements in a molecule and determining the structure of a

compound by observing its fragmentation.

Other uses include quantifying the amount of compound in a

sample. This picture is depicting the M s procedure.

The sample comes in from the left, goes through these

steps before it's detected and amplified to the right and

then it's recorded by a chart.

This picture is depicting the L C procedure.

Same thing Sample comes in from the left, goes through

these steps before it's detected on the right.

In this case, the detection is shown in peaks on

a computer, the procedure done within the paper.

First, the samples were prepped using an automated micro lab

star system from Hamilton Company.

Second, the step was done to remove proteins, disassociate small

molecules bound to protein or trapped in the precipitated protein

matrix, and to recover chemically diverse metabolites.

Third, the extracted samples are separated into these five fractions

where R P stands for reverse phase.

The ion mode is for electro spray ionization, for they

removed the organic solvents by placing them briefly on a

turbo vac buys a mark five.

They restored overnight.

Six. The samples were ran through both M S and

L C machines and data was collected.

Seven. The solvents are compatible with each of the four

methods, and each reconsider situation solvent contained a series of

standards that fixed concentrations.

Eight. This step is called bulled pools.

Biomechanics metabolites were quantified as area under the curve and

normalized to protein levels for each sample.

To account for potential differences in the amount of material

present in each sample.

The second experimental method used to collect the data was

fly scape.

Fly scape is split into three tiers first, as the

presentation tier consists of the plug in for site Escape

66 The logic here is a standard Web service that

provides access to the data stored in the database and

specific logic, such as deriving the metabolic grass or performing

enrichment analysis.

The data tier is represented by an SQL Server database

that is accessed by the Web service.

The databases you need to fly escape.

The major objects in the database are compounds, reactions, enzymes,

jeans, pathways and their relationship.

The data is returned and easily translate into job objectives.

In both the presentation and logic here, this combination allows

for easy extension of the services produced by fly scape.

The procedure for this in the paper was first flies.

Cape supports four types of network graphs one compound to

compound reaction, three compound gene and four compound reaction enzyme

gene. Second, for the case of the study, notes of

the same type are never connected to each other.

Edges between compound and reaction notes are directional to signify

the reaction substrates and products and third fly scape supports

to queries the user.

Can one supply a list of genes and or compounds,

or to select one of the canticle metabolic pathways?

Now that we understand the procedures used, let's look more

deeply at the results.

First, we have to a.

The normalize metabolic levels were clustered by compound as seen

in the rose and by biological replicates in the column.

The names are also colored according to their metabolic classes,

and the key for these can be found in the

bottom right corner of the figure.

The heat map is also split down the middle and

re fed flies are on the left with the fast

it flies on the right.

The setup is going to be the same as seen

and to see.

Of the 61 change metabolites, 31 metabolites were increased, while

30 decreased within the re fed flies.

The compounds that were higher and re fed flies reflect

increases in glucose availability, metabolism utilization for post translational modification

and storage into glycogen.

Fasted flies showed an increase in metabolites such as glutamine,

alpha kilo obliterate tyrosine and medium chain fatty acids, which

indicates energy coming from the higher metabolism of compounds like

fatty acid, branch chain, amino acid and ketone bodies.

Fast it flies also had changes in peering and promoting

CA Tabal is, um, the rear cycle and the Argentine

and protein metabolism.

We found that while metabolites change rapidly upon feeding in

both the head body tissue, the response of each to

fasting and eating are distinct body show, a slow response

to nutrient availability to be as shown in the key.

Salmon colored hexagons are representing compounds that were weren't measured

by fly escape.

The statistical test used was the wildest T test, which

is a two sample location test which is used to

test the hypothesis that the two populations of equal means

and it gave an FDR less than 20.1.

FDR stands for false discovery rate.

This means that less than 10% of the jeans tested

are not false positives.

Fly Escape reveals a change in the T.

C, a cycle that is easy to appreciate.

It is fueled by glucose and re fed flies and

buy glutamine through alpha keto gluten rate and fast It

flies. One looked at together.

This data provides evidence of a shift from beta oxidation

slash fatty acid metabolism and ketosis during the fasted state

to increase glucose utilization after re feeding now for to

see as mentioned previously, it has the same set up

as Figure two A.

But now it just represented the data found in the

head. Most of the metabolites in the head decreased with

fasting and comparatively to the bodies of fruit flies.

There was a higher glucose availability and higher amounts of

glucose related products.

This increasing glycogen intermediate suggests higher glucose demands and heads

this data together with the elevated levels of glucose, methylene

reflects higher octave stress and fasted fly heads compared to

bodies where no change was observed.

Okay, figure two d again the same set up as

we saw and figure to be here increases in Pintos

phosphate metabolites like K E G G and D M

e, all of which indicates a differential and likely increased

use of glucose.

Another suggestion that the glucose demand was higher in the

heads arose from the increasing glycogen intermediates.

Finally, higher levels of T C A cycle metabolism metabolites

and the Peruvian acetyl COA Intermediate Acetyl phosphate suggests that

carbon utilization into the T.

C A cycle may occur primarily from Peru, derived from

carbohydrate sources in the refried state and the heads of

fruit flies.

Lastly for Figure two e, it's depicting a Venn diagram

that shows the overlap and the metabolic shifts between the

bodies, lavender of fasted and re fed flies and the

heads green of the flies.

In conclusion, while both heads and bodies showed an increase

in energy availability with re feeding, their metabolic profiles were

largely non overlapping, with higher glucose metabolism changes in the

head. Finally, we'll go to the discussion.

This paper will help lay the groundwork for future studies

as it confirms that the state of one's diet, whether

it's fasting, satiety or a high sugar diet, alters the

way in which the metabolic pathways are assigned their nutrients.

The information gained will help in future studies that attempt

to understand the potential effects of specific metabolite transition on

the physiology of the brain, as well as their role

in the development of brain conditions associated with diet such

as neuro degeneration, depression and seizures.

Some caveats of this study include the researchers sampling the

whole heads of the flies instead of studying only the

brains. This could potentially reduce the reliability of the data

that was collected.

Another example would be from when they quantify the metabolite

data they collected from the L.

C. M s procedure.

They quantify them as areas under the curve, which can

introduce error as the ways used to process the area

either provide over under estimates many molecules such as Chunlin,

Anna Segal, serotonin, glutamine and G A B A.

That increase in re fed flies and aspirate eight and

N A that were higher and fasted animals have known

or surfacing roles in helping to modulate cellular behavior.

This data could be used to help speculate on what

the paper is searching for changes in metabolites having informational

value that affect the function of the brain.

However, on the other hand, this data also supports that

the metabolites levels could instead be mediated by circuit and

synaptic levels and that are modulated by hormone levels, organ

communication and neural peptides.

Signaling future research that could be done to help decipher

between the two results would be using cell attached voltage

clamps in order to see if the changes were brought

about by synaptic changes or not.

I do believe that the interpretations laid out in this

paper as the fly Scapin L CMS data I discussed

earlier was further solidified with studying the abundance of RNA.

When doing so, the researchers found that several nutrient transporters

and metabolic enzymes change rapidly in the brains of fasted

and re fed flies.

These results are good indication that metabolites as messengers and

transducers of environmental information, plays some sort of role and

complex behavior and have provided substantial base word for further

studies to build off of as expressed earlier for the

research is now need to confirm which path light metabolite

levels are synaptic and circuit levels is responsible for the

complex behavioral changes in the brain.

If I were to join the lab, this would be

the point where I begin to further the understanding of

these complex behavior.

These are my references, and does anybody have any questions

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