| | 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 |
| | |
| | |
| | |
| | -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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