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