Screening Antimalarial Drugs for Modulation of Aggressive Behaviour in Drosophila Melanogaster

 TABLE OF CONTENTS

DECLARATION ................................................................ ; ........................................................... 2

APPROVAL ................................................................................................................................... 3

DEDICATION ......................................................................................................................... · ....... 4

ACKNOLWEDEMENTS ......................................................... : ..................................................... 5

LIST OF ACRONYMS ........................................................................................ ~ ......................... 6

LIST OF FIGURES ...................................................................................................................... 10

LIST OF TABLES ........................................................................................................................ 10

ABSTRACT .................................................................................................................................. 11

CHAPTER ONE ........................................................................................................................... 12

1.0 INTRODUCTION .................................................................................................................. 12

1.1 Background ........................................................................................................................... 12

1.2 Problem statement. .................................................................................................................. 13

1.3 Objectives ............................................................................................................................... 13

1.3.1 Broad objective .................................................................................................................... 13

1.3.2 Specific objectives ............................................................................................................... 14

1.4 Research questions/ Hypothesis .............................................................................................. 14

1.5 Justification of the study ......................................................................................................... 14

CHAPTER TWO ............................................................ : ............................................................. 15

2.0 LITERATURE REVIEW ....................................................................................................... 15

2.1 Aggression .............................................................................................................................. 15

2.2 Biological signals and aggression .................................................................................... : ...... 16

2.3 Anti-malarias and aggression .................................................................................................. 17

CHAPTER THREE ...................................................................................................................... 18

3.0 MATERIALS AND METHODS ............................................................................................ 18

3.1 Area of study ........................................................................................................................... 18

3.2 Study design ..... : ...................................................................................................................... 18

3.3 Materials .............. · ................................................................................................................... 18

3.3 .1 Fly starch .............................................................................................................................. 18

3.3.2 Chemicals and reagents: ...................................................................................................... 18

3.3.3 Preparation of fly food ........................................................................... , ............................. 18

3.3.4 Drugs: ................................................................................................................................... 19

3.3.5 Equipment ............................................................................................................................ 19

8

3.4.0 Methods ................................................................................................................................ 19

3.4.1 Fly preparation for expeliment. ........................................................................................... 19

3.4.3 Drug administration ............................................................................................................. 20

3.5.0 Pharmacological treatments ................................................................................................. 21

3. 5.1 for serotonin ( 5HT) .............................................................................................................. 21

3.5.2 for Dopamine·············:········································································································· 21 3.5 .3 for Octopamine .................................................................................................................... 22

3.6 Anti-malaJial interference with Neurotransmitter pathways .................................................. 22

3.7 Expected outcomes ................................................................................................................. 22

CHAPTER FOUR ......................................................................................................................... 23

4.0 RESEARCH RESULT FINDINGS ........................................................................................ 23

4.1 Behavioral changes in flies treated with antimalarials ........................................................... 23

4.2 Activities of antimalarials on mediators of aggression ........................................................... 25

4.2.1 FOR OCTOP AMINE ........................................................................................................... 25

4.2.2 FOR DOPAMINE ................................................................................................................ 27

4.2.3 FOR SEROTONIN .............................................................................................................. 29

4.3 RELATIONSHIP BETWEEN AGE AND ANTIMALARIAL DRUG INDUCED AGGRESSIVE BEHAVIOR ........................................................................................................ 32

CHAPTER FIVE .......................................................................................................................... 34

5.0 DISCUSSION ......................................................................................................................... 34

5.1 Aggression in flies treated with antimalalials ......................................................................... 34

5.2 Activities ofantimalalials on mediators of aggression ........................................................... 35

5.2.1 FOR OCTOPAMINE ........................................................................................................... 35

5.2.2 FOR DOPAMINE (da) ........................................................................................................ 36

5.2.3 FOR SEROTONIN (5HT) ................................................................................................... 36

CHAPTER SIX ............................................................................................................................. 39

6.0 CONCLUSION AND RECOMMENDATION ...................................................................... 39

REFERENCES ............................................................................................................................. 40

APPPENDICES ............................................................................................................................ 48

APPENDIX 1: TIME FRAME ..................................................................................................... 48

APPENDIX 2: BUDGET ............................................................................................................. 49

APPENDIX 3: BENCH WORK PICTURES ............................................................................... 50

ABSTRACT Background: Aggressive behavior is crucial to survival and reproduction in organisms and this can be affected by environmental factors. Antimalarial are commonly used in medical practice, and it's possible that they would interact with modulators of aggression to influence behavior since some have been shown to cause neuro-damage. Objective: This was to evaluate common antimalarial drugs for modulation of aggressive behavior in Drosophila melanogaster model. Materials and Methods: Drosophila melanogaster specie (Wll8) was used in this study. Flies were divided into male and female and experiments were conducted on adult, middle and juvenile age groups. Flies were exposed to chloroquine (0.0025 mg/ml), quinine (0.000135 mg/ml), Fansidar (0.0025 mg/ml), Artesunate (0.0003 mg/ml), and Artemether lumefantrine (0.0003 mg/ml). These were later on exposed to neurotransmitter modulators i.e. octopamine (stimulatory-clonidine (I mg/ml); inhibitory-promethazine (0.00025 mg/ml)), dopamine (stimulatory-L-dopa (0.001 mg/ml); inhibitor-haloperidol (0.0001 mg/ml), serotonin (stimulatory-fluoxetine (0.0002 mg/ml), inhibitory-cyproheptadine (0.00004 mg/ml)). Data was recorded in triplicate and analyzed in MS Excel. Infonnation was presented in mean± SEM and significance at 95% was considered. Results and Discussion: The study showed that Artesunate had the highest effects of aggression in male Drosophila melanogaster flies while Quinine and Chloroquine were associated with low effects and Artemether lumefantrine was associated with low level of aggression in female flies. These observations would have been due to their interaction with neurotransmitter release which is essential for aggressive behavior. Fansidar and Artemether-lumefantrine acted synergistic to octopaminergic stimulation in both males and females respectively. Artesunate antagonized actions of promethazine by leading to increased aggression especially in male flies. Fansidar and Artemether-lumefantrine acted synergistic to dopaminergic stimulation while Artesunate antagonized dopaminergic inhibition, showing that it plays a cmcial role in aggression. Serotonin leads to decreased aggression and Fansidar showed antagonist activity in males while in females this was in Artemether Lumefantrine. Artesunate showed strong inhibitory activity on serotonin release, thus leading to increased aggression. In the age groups, aggression by Artesunate was highest in adult male and female flies and this raises major phannaceutical concerns. In the juveniles, Fansidar and Artesunate showed significant aggression, thus showing implications in neural development. Conclusion and Recommendation: Molecular mechanism on actions of Artesunate and Fansidar on modulation of neurotransmitter release need to be investigated further to gain much clear insight