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SEADAP LESSON OUTLINE
Lesson Title Description
1 Acute Activity
(Stereotypy)
C-shapes increase during acute
drug exposure
2 Repeated Activity
(Stereotypy)
Tolerance or sensitization to C-
shapes occurs during repeated
drug exposure
3 Acute Activity
(Motility)
Locomotion (grid crossings)
changes during acute drug
exposure
4 Natural Anxiety Greater time in dark indicates
greater anxiety
5 Dependence (Anxiety
and Drug Withdrawal)
Greater time in dark following drug
withdrawal indicates greater anxiety
6 Dependence
(Depression and Drug
Withdrawal)
Decreased motility/activity following
drug withdrawal indicates
depression
7 Reward and
Reinforcement
(Conditioned Place
Preference)
Environmental cues associated with
drug reward facilitate drug
addiction
8 Drug-Drug
Interactions
Combinations of caffeine/sugar,
caffeine/alcohol, etc. can produce
additive, synergistic or sub-additive
effects on activity and reward
Teacher’s Guide: Acute Sterotypy--50 minutes total
Goal: To demonstrate that drugs of abuse affect activity following acute exposure. Stimulants such as nicotine and caffeine will produce increased activity following acute exposure and this effect in humans is due to increased dopamine transmission in area of the brain called the limbic system. The C-shapes observed in this experiment represent a stereotyped behavior. Stereotyped behaviors are defined as repetitive, meaningless movements and are commonly observed when stimulants are administered to rodents.
Part 1: Vocabulary--approximately 5-10 minutes
Word Definition
Planarian A kind of flatworm, and the stars of this experiment!
Solute The minor component in a solution, dissolved in the solvent
Solvent The major component in a solution, what the solute is dissolved in
Solution A liquid mixture where the solute is dissolved in the solvent (Ex: Salt Water, when you have dissolved salt in water, Salt here is acting as the solute and water is acting as the solvent
Concentration The amount of dissolved solute per unit volume of solution
Acute A response of rapid onset and/or short duration
Stereotypy Repetitive or ritualistic movement (Ex: Body rocking, C-Shaped Movements)
Part 2: Observations --approximately 10 minutes
- Place the planarian in a petri dish of spring water using the disposable pipettes. Using the microscope, observe the planarian under the microscope.
- Draw a picture in the circle below of what you saw under the microscope as well as list your observations.
Under observations students should note color, movement, size, shape, etc. It is recommended that the microscopes are set up for the students.
0.1 mM 1.0 mM 5.0 mM 10 mM
Trial 1 4 8 18 22
Trial 2
Trial 3
Average (Trials 1-3)
**Note: these are the values obtained with our own experimental procedure, they may vary slightly from classroom to classroom. It is recommended that you test these on your own first!
Fill in the graph with your results below (use the averages).
CAFFEINE
- Label a petri dish with “.01 mM Caffeine”.
- Place your planarian on a petri dish with .01 mM of Caffeine.
- How many times does it exhibit C-Shaped behavior in 2 minutes (when the planarian moves its body into a C-Shape and then back to normal)?
- When finished, place the planarian back in a petri dish of spring water.
- Repeat steps 1-4 an additional 2 times with different planarians.
Repeat the above with Caffeine concentrations of 1 mM, 5 mM and 10 mM. Record your findings in the table below. Note: with a concentration of 10 (and maybe even 5) dose of drug may become so high that it is toxic to the planarian. If you do not want this to occur in your classroom just alter the experiment to eliminate that high dose.
0.1 mM 1.0 mM 5.0 mM 10 mM
Trial 1 2 3 9 13
Trial 2
Trial 3
Average (Trials 1-3)
**Again, these values are results obtained for our own experimental procedure and will vary!
Fill in the graph with your results below. Example graphs for each substance can be found above
ALCOHOL
- Label a petri dish with “.01 mM Caffeine”.
- Place your planarian on a petri dish with .01 mM of Alcohol.
- How many times does it exhibit C-Shaped behavior in 2 minutes (when the planarian moves its body into a C-Shape and then back to normal)?
- When finished, place the planarian back in a petri dish of spring water.
- Repeat steps 1-4 an additional 2 times with different planarians.
Repeat the above with Alcohol concentrations of 1 mM, 5 mM and 10 mM. Record your findings in the table below. Note: with a concentration of 10 (and maybe even 5) dose of drug may become so high that it is toxic to the planarian. If you do not want this to occur in your classroom just alter the experiment to eliminate that high dose.
0.1 mM 1.0 mM 5.0 mM 10 mM
Trial 1 1 8 1 1
Trial 2
Trial 3
Average (Trials 1-3)
**Note, results may vary
What methods did we use to test our hypothesis? Placing planarians into four different concentrations of the drug solution and counting the number of times they exhibited the C-shape movement in a five minute interval
What were our results? For both Nicotine and Caffeine , increased substance concentration is associated with increased movement (C-Shaped Curves) For Alcohol there is a sharp increase in movement at 1 mM but the effect most disappears with a smaller or larger concentration. Sucrose is the control condition and as a result there was no effect of increasing concentration on movement.
What conclusions can we draw from our results? Nicotine and caffeine exposure will increase movement Alcohol will increase movement at a certain concentration (1 mM) but have little/no effect at other concentrations Sucrose exposure have no effect on movement
Part 5: Discussion Questions--approximately 10 minutes
- What patterns do you notice as the concentrations increase? Generally, as concentrations increase, there is increased movement (Note: Sucrose is a control so no changes are observed in this condition)
- Is there a relationship between substance concentration and planarian movement? For every substance but sucrose, generally the higher the concentration, the greater the movement
- How can you tell that an increased substance concentration is having a greater effect on the planarian? You can tell that increasing substance concentration is having a greater effect on the planarian because the planarian moves more with higher concentrations. This might suggest that these substances have the potential to have physical effects on the organisms that ingest them.
- Part of what makes planarians so cool is that they have a brain and a central nervous system that is a lot like ours. What might this mean about how these substances (nicotine, caffeine, alcohol, etc.) affect our own bodies?
If these substances are having physical effects on planarians, then they probably will have physical effects on humans as well. Also, in general, the more of a substance you ingest, the greater the physical effects will be.
Part 3: Experiment
How is a planarian affected by nicotine, caffeine, alcohol, and sucrose? Note: each group will be assigned one of the above substances to investigate
NICOTINE
- Label a petri dish with “.01 mM Nicotine”.
- Place your planarian on a petri dish with .01 mM of Nicotine.
- How many times does it exhibit C-Shaped behavior in 2 minutes (when the planarian moves its body into a C-Shape and then back to normal)?
- When finished, place your planarian in a petri dish with spring water.
- Repeat steps 1-4 an additional 2 times with different planarians.
Repeat the above with Nicotine concentrations of 1 mM, 5 mM and 10 mM. Record your findings in the table below.
0.1 mM 1.0 mM 5.0 mM 10 mM
Trial 1
Trial 2
Trial 3
Average (Trials 1-3)
Fill in the graph with your results below (use the averages!).
CAFFEINE
- Label a petri dish with “.01 mM Caffeine”.
- Place your planarian on a petri dish with .01 mM of Caffeine.
- How many times does it exhibit C-Shaped behavior in 2 minutes (when the planarian moves its body into a C-Shape and then back to normal)?
- When finished, place your planarian back in a petri dish with Spring Water.
- Repeat steps 1-4 an additional 2 times with different planarians.
Repeat the above with Caffeine concentrations of 1 mM, 5 mM and 10 mM. Record your findings in the table below.
0.1 mM 1.0 mM 5.0 mM 10 mM
Trial 1
Trial 2
Trial 3
Average (Trials 1-3)
Fill in the graph with your results below (use the averages!).
SUCROSE
- Label a petri dish “.01 mM Sucrose”
- Place your planarian on a petri dish with .01 mM of Sucrose.
- How many times does it exhibit C-Shaped behavior in 2 minutes (when the planarian moves its body into a C-Shape and then back to normal)?
- When finished, place your planarian back into a petri dish with spring water.
- Repeat steps 1-4 an additional 2 times with different planarians.
Repeat the above with Sucrose concentrations of 1 mM, 5 mM and 10 mM. Record your findings in the table below.
0.1 mM 1.0 mM 5.0 mM 10 mM
Trial 1
Trial 2
Trial 3
Average (Trials 1-3)
Fill in the graph with your results below (use the averages!).
Part 4: Scientific Method Recap
What question were we trying to answer with this experiment?
What might have been our hypothesis?
What methods did we use to test our hypothesis?
What were our results?
What conclusions can we draw from our results?
Part 5: Discussion Questions
- What patterns do you notice as the concentrations increase?
Lesson 2: Repeated Stereotypy
Goal: To demonstrate using planarians that drugs of abuse produce changes in activity during repeated exposure. Repeated exposure to a substance may produce a diminished biological response, which is defined as tolerance, or an augmented biological response, which is defined as sensitization. Repeated exposure is also defined as chronic exposure. The pattern of chronic exposure can mimic addictive use in humans (i.e., binge-like administration on a daily basis) or recreation use (i.e., periodic use that may be only on the weekends or once a month).
Part 1: Vocabulary
Planarian A kind of flatworm, and the subject of this experiment
Solute The minor component in a solution, dissolved in the solvent
Solvent The major component in a solution, what the solute is dissolved in
Solution A liquid mixture where the solute is dissolved in the solvent (Ex: Salt Water, when you have dissolved salt in water, Salt here is acting as the solute and water is acting as the solvent
Concentration The amount of dissolved solute per unit volume of solution
Stereotypy Repetitive or ritualistic movement (Ex: Body rocking, C-Shaped Movements)
Tolerance a drug or concentration of drug loses its effectiveness with chronic exposure
Sensitization an exaggerated response during chronic exposure that sometimes occurs with drugs of abuse
Dependence the state of being psychologically or physiologically dependent on a drug after a prolonged period of use
Physical Dependence
dependence that involves persistent physical–somatic withdrawal symptoms (e.g., fatigue and delirium tremens)
Psychological Dependence
dependence that involves emotional–motivational withdrawal symptoms (e.g., dysphoria and anhedonia)
Part 2: Observation
- Examine the planarian under a microscope and draw what you see. List your observations below.
Under observations students should note color, movement, size, shape, etc. It is recommended that the microscopes are set up for the students.
Part 3: Experiment
How is a planarian affected by repeated exposure to nicotine, caffeine, and alcohol? Note: each group will be assigned one of the above substances to investigate
This experiment is testing sensitization versus tolerance. It is very hard to predict which substances will result in either scenario, as it will differ from planarian to planarian. Sensitization will occur when the response becomes more and more robust with additional trials, and tolerance occurs when the response becomes smaller and smaller with additional trials. Both sensitization and tolerance occur as a result of chronic exposure, which is why three trials are necessary for each planarian tested. You can also vary the number of times the planarians are exposed to the drugs or the duration of the withdrawal interval. For example, you might try exposing the planarians to the drug for 3 straight days using a withdrawal interval of 24 hours. The long withdrawal interval would mimic recreational drug use in a human whereas the shorter withdrawal intervals of 8 minutes would mimic a binge-type exposure that is characteristic of addictive drug abuse.
NICOTINE
- Prepare one petri dish with a 1 mM solution of nicotine and one petri dish filled with spring water and place both on a plain white piece of paper. You will want to test out the drug concentration under your own laboratory conditions. We recommend starting with 1 mM. You may need to adjust the concentration slightly down (e.g. 0.1 mM) if the planarians die with repeated exposure or slightly up ( e.g. 5 mM) if the response is not robust enough.
- Using a pipette, remove the planarian from its home jar
- Using a pipette, place the planarian into the nicotine solution
- For 2 minutes, count the C-shapes and record this number in the table below
- Using a spatula, place the planarian into the spring water for 8 minutes
- While you wait, exchange data from your first trial with the other groups
- Repeat steps 3-6 two more times
- Return the planarian to its home jar
- Exchange information from your third trial with the other groups
- Fill in the graph with your results below
Trial 1 2 3
# of C-shapes
ALCOHOL
- Prepare one petri dish with a 0.1% solution of alcohol and one petri dish filled with spring water and place both on a plain white piece of paper. You will want to test out the drug concentration under your own laboratory conditions. We recommend starting with 0.1%. You may need to adjust the concentration slightly down (e.g. 0.01%) if the planarians die with repeated exposure or slightly up ( e.g. 1%) if the response is not robust enough.
- Using a pipette, remove the planarian from its home jar
- Using a pipette, place the planarian into the alcohol solution
- For 2 minutes, count the C-shapes and record this number in the table below
- Using a spatula, place the planarian into the spring water for 8 minutes
- While you wait, exchange data from your first trial with the other groups
- Repeat steps 3-6 two more times
- Return the planarian to its home jar
- Exchange information from your third trial with the other groups
- Fill in the graph below with your results
Trial 1 2 3
# of C-shapes
Part 4: Scientific Method Recap
What question were we trying to answer with this experiment? Which drugs cause a planarian to exhibit sensitization or tolerance over chronic use?
What might have been our hypothesis? X drug(s) cause sensitization while X drug(s) cause tolerance.
What methods did we use to test our hypothesis? We exposed planarians to the same drug repeatedly and measured the number of C-Shaped curves that each planarian exhibited in response to the chronic drug use.
What were our results? Some planarians experienced sensitization while others experienced tolerance.
