Lifespan Extension and Locomotor Activity of Beverage Ingredients in Drosophila Model
- 주제(키워드) lifespan , locomotor , beverage , drosophila
- 발행기관 고려대학교 대학원
- 지도교수 서형주
- 발행년도 2017
- 학위수여년월 2017. 2
- 학위구분 석사
- 학과 대학원 의생명융합과학과
- 세부전공 식품영양과학
- 원문페이지 121 p
- 실제URI http://www.dcollection.net/handler/korea/000000072355
- 본문언어 영어
- 제출원본 000045900894
초록/요약
This study was aimed to investigate the effect of functional drinks and the principal ingredients of functional drinks on the relationship among lifespan, antioxidant activity and behavior pattern using Drosophila melanogaster, called as a fruit fly. The survivorship was evaluated by measuring mean lifespan (MLS) and median survival (lethal time, LT50). Estimation of antioxidant activity was determined by measuring enzymatic activities, such as superoxide dismutase (SOD) and catalase (CAT), and glutathione (GSH) in addition to quantification of expression mRNA level related with the antioxidant activity (Sod1, Sod2, Cat and GstD1). Meanwhile, behavior pattern in fly model was observed by various locomotor activities. In addition, expression levels of mRNA associated with locomotor activity (Dop1R1, Rdl, GABAB-R1 and GABAB-R2 gene) were investigated for elucidation of behavior mechanism. Finally, various genes from the D. melanogaster were collected, and the genes were further analyzed by next generation sequencing (NGS) and differentially expressed gene (DEG) analyses, to identify the relationship of three factors evaluated. All assays were compartmentally performed by treating three functional drinks (vitamin drink, VD; taurine drink, TD; caffeine drink, CD) or by three main ingredients (vitamin C, taurine and caffeine). The result of lifespan showed that vitamin C supplementation did not show significant difference compared with normal group. In contrast, 1.5% taurine and 0.025 and 0.05% caffeine supplementation led to significant decrease of lifespan. In the other hands, longevities of the drinks supplementation were as follows in decreasing order: normal group > VD > CD > TD. In enzyme activity, 1% vitamin C supplementation showed the significantly increase of activities in CuZnSOD (19.88 U/mg protein), MnSOD (0.80 U/mg protein), CAT (98.53 mM/mg protein) and GSH (0.35 nM/mg protein). In result of gene expression level, 1% vitamin C supplementation showed the highest level of GstD1 expression. Furthermore, 0.05% caffeine consumption showed the highest of Dop1R1 gene expression. In contrast, 1.5% taurine consumption showed the highest of Rdl, GABAB-R1 and GABAB-R2. DEG analysis showed the interaction among genes related with antioxidant activity, hormone activity and locomotor behavior in D. melanogaster exposed 3 ingredients (vitamin C, taurine and caffeine). In behavioral assays, taurine and caffeine treatments significantly influenced daytime and nighttime activity (p < 0.05). In total movement activity, 0.05% caffeine supplementation and CD showed the highest activity at the subjective nighttime. In contrast, 1.5% taurine supplementation and TD showed the lowest total activity at the subjective nighttime and daytime. Climbing abilities at nighttime and daytime were significantly decreased in 1.5% taurine supplementation and TD group. Various behavior factors i.g. total distance, velocity, movement etc, in taurine and caffeine supplementation showed different locomotor activities compared to normal group. These results were showed that lifespan, antioxidant and behavior were closely related. Furthermore, vitamin C acted as a body enhancer that maintains the stability of the body rhythm and increases anti-oxidative activity. In contrast, taurine and caffeine are neuromodulators that induce behavioral inhibition and arousal. Change of behavior and locomotor activities might affect to decrease lifespan.
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ABSTRACT i
CONTENTS iv
LIST OF TABLES vii
LIST OF FIGURES viii
General Introduction 1
Chapter 1. 7
I. INTRODUCTION 8
II. Material & Methods 11
2.1. Chemicals 11
2.2. Drosophila melanogaster stock and rearing condition 11
2.3. Survivorship study of D. melanogaster 12
2.4. Estimation of antioxidant activity by enzymatic bioassay for D. melanogaster 14
2.5. Differentially Expressed Gene (DEG) analysis 18
2.6. Statistical analysis 21
III. Results and Discussion 22
3.1. Effect of vitamin C, taurine and caffeine on lifespan of D. melanogaster 22
3.2. Effect of commercial drinks on lifespan of D. melanogaster 25
3.3. Effect of vitamin C, taurine and caffeine on survival rate of D. melanogaster induced oxidative stress with hydrogen peroxide 28
3.4. Effect of vitamin C, taurine and caffeine on enzyme activity associated with antioxidant in D. melanogaster 30
3.5. Effect of vitamin C, taurine and caffeine on gene expression associated with antioxidant in D. melanogaster 34
3.6. Effect of vitamin C, taurine, and caffeine on gene expression associated with behavior of D. melanogaster 37
3.7. Differentially expressed gene (DEG) analysis of D. melanogaster 41
Chapter 2. 54
I.INTRODUCTION 55
II. Material & Methods 57
2.1. Chemicals 57
2.2. Drosophila melanogaster stock and rearing condition 57
2.3. D. melanogaster locomotor activity 58
2.4. Statistical analysis 62
III. Results and Discussion 63
3.1. Effect of vitamin C, taurine, and caffeine on circadian rhythm and total movement of D. melanogaster 63
3.2. Effect of commercial drinks on circadian rhythm and total movement of D. melanogaster 70
3.3. Effect of vitamin C, taurine, and caffeine on climbing ability of D. melanogaster 74
3.4. Effect of commercial drinks on climbing ability of D. melanogaster 77
3.5. Effect of vitamin C, taurine, and caffeine on open field assay (locomotion) of D. melanogaster 80
3.6. Effect of commercial drinks on open field assay (locomotion) of D. melanogaster 86
CONCLUSION 92
ABSTRACT IN KOREAN 95
References 98
