Category ANIM L01 Toward Further Understanding in Human Addiction Using C.


Abstract Objective(s):

-Observe behavior of C. elegans when they must choose between the

drug and a substance that they are normally attracted to - NaCl.

-Study the neurotransmitters that may have been responsible for their

potential behavior.

-Compare the different mutants to each other and analyze the different

patterns that each of them holds.

-Find a connection between the worms' behavior and human addiction

-Mutants include tph-1 and dat-1

-N2 is control

-Dat-1: encodes plasma membrane dopamine transporter; “is predicted

to regulate dopaminergic neurotransmission”

-Tph-1 gene encodes tryptophan hydroxylase, which is “the enzyme

that catalyzes the rate-limiting first step in serotonin biosynthesis.”

-SAM-e: antidepressant; naturally found in human body

-Benadryl: allergy med; drug that reduces anxiety symptoms but causes

it in others

-Caffeine: central nervous system stimulant and the world's most widely

consumed psychoactive drug

-Nicotine is a stimulant drug and most commonly found in cigarettes

-All drugs will be tested separately, and all of them will be tested

alongside a well-known attract to C. elegans: NaCl.

Part 1 - Testing various concentrations of the drugs to determine which

is the best to use. Dilute all drugs to 5, 10 and 15 mM concentrations.

Only about 1 mL of each is needed. Drugs are made from pill to liquid

form using water and sodium azide. Petri dishes are cut into four

sections - two for test "T" and two for control "C". Each substance -

drugs for test and 100 mM NaCl (for control) are placed onto the plates

accordingly. Only N2 worms were used as they were put onto the

middle of the plate after the drugs and control had dried in the middle

of each of their sections. After one hour (trial time), the number of

worms were counted and recorded for each concentration/drug

combination, either under "T" for test, "C" for control, "M" for middle, or

"E" for elsewhere. Water and 3.75 M NaCl were also used separately as

tests alongside the 100 mM NaCl.

Part 2 - Testing mutants against drug concentrations determined in

Part 1.

Concentrations found based on most distinctive results: 10 Mm

caffeine, 5 mM benadryl, 10 SAM-e, and 5 mM nicotine.

Best timing for chemotaxis - about two hours and recount morning after

to ensure accuracy of worms and that they stopped moving

Conduct chemotaxis assay with the new concentrations but now with the

mutants, not only the N2 worm. Be sure to put 5-10 worms on the

middle on each plate when chemotaxing them.

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