1. nNOSI and 6OHDA Treatments Illicit Different Motor Deficiency Symptoms in Embryonic Zebrafish
Presented by: Connor Culley, Derek Emerson, and Chris Baker
BI-420W-01 Seminar Research Presentation
Biology Department
Virginia Military Institute

2. Introduction Estrogen’s (E2) is a steroidal signaling hormone
Function: neuron protection .
Upstream regulator of nitric oxide synthesis (NOS).
Dopamine (DA) is a neurotransmitter involved in motor function
6OHDA is a neurotoxin that causes dysfunction in DA producing neurons in zebrafish (Parng et al., 2007)
nNOSI is an enzyme responsible for the inhibition of the synthesis of nitric oxide (NO) in nervous tissue
NO is a gaseous signaling molecule involved in motor control (Lorenc-Koci and Czarnecka, 2013)

3. Hypothesis
If embryonic zebrafish are exposed to either nNOSI or 6OHDA, then they will produce the same listless PD-like symptoms. E2 co-treatment will rescue the fish from these neurological symptoms

4. Zebrafish Model Why Zebrafish? Optically transparent CNS and organs.
Known genome.
Easy maintenance of survival.
Readily absorb chemicals.
Small size conducive to larger sample size.
Simple, cheap, rapidly reproducible biological model.
An established model for the study of motor deficits with PD-like symptoms (Flinn et al., 2008)

5. Materials 6-day old Casper zebrafish Dissecting microscope
Pipets and probes
6-OHDA (500 µM), nNOSI (50 µM) , ERS, E2 (100 µM)
Sterilized 6-well plates
Incubator

6. Methods Exp #1: nNOSI (10, 25, 50 µM) dose response study
Exps #2 – 6: treatments:
ERS
nNOSI
6-OHDA
E2 + nNOSI
E2 + 6OHDA

7. Methods
Motor behavior analyses (% of population demonstrating deficits 24 hrs after treatment)
Listless – no spontaneous swimming, laying on side, now swimming response to probe
Dyskinesia – rapid sporadic, twitchy/spastic swimming
Data analysis
Data were analyzed for significant differences either by a z-test for two population proportions or for multiple proportions using a chi-square contingency table test followed by a Marascuilo's post hoc analysis

8. Directions: insert error bars (±SEM); do stats for multiple proportions; write figure legend
Results
Figure 1.

9. Results
Directions: cull out just 24 hor data & add nNOSI results; insert error bars (±SEM); do stats for multiple proportions; write figure legend
Figure 2.

10. Conclusions
Directions: finish, 2-3 points

11. Future Studies

12. References Flinn et al., 2008 Lorenc-Koci and Czarnecka, 2013
Parng et al., 2007
Directions: complete citations

13. Acknowledgements