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研究生中文姓名:瑪格
研究生英文姓名:Maigan Espinili Maruquin
中文論文名稱:高溫對於石斑魚腸道先天性免疫反應之影響
英文論文名稱:Impact of High Temperature on the Innate Immune Response in Grouper Intestine
指導教授姓名:邱品文
口試委員中文姓名:教授︰張本恆
教授︰呂明偉
副教授︰陳媺玫
助理教授︰邱品文
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:水產養殖學系
學號:10633059
請選擇論文為:應用型
畢業年度:108
畢業學年度:107
學期:
語文別:英文
論文頁數:49
中文關鍵詞:基因表現,免疫刺激,腸道先天免疫,溫度基因表現免疫刺激腸道先天免疫溫度
英文關鍵字:gene expressions,intestinal innate immunity,immunostimulation,temperaturegene expressionsintestinal innate immunityimmunostimulationtemperature
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摘要
我們分析高溫對於點帶石斑魚腸道先天免疫反應的影響。將魚苗置於三個不的水溫環境下:28℃(對照組)、28℃每日增溫1℃至33℃(熱適應性處理組)、及28℃急遽增溫至33℃(熱急速處理組)。7日後,每組魚均分別注射LPS、CpG ODN和Poly I:C。於免疫刺激1、3、5日後收集腸道組織樣品(前、中、後段)以分析mRNA的相關先天性免疫相關基因的表現。分析結果顯示高溫及升溫模式對於上述三種免疫刺激物在腸道誘導免疫基因表現具有明顯的影響效應,且影響效應因腸道區域而異。結果更進一步顯示高溫處理(熱急速及熱適應性處理)對於石斑魚腸道CpG ODN免疫刺激反應最不具負面影響,甚至可提升特定免疫基因的誘導表現。
Abstract
We conducted an investigation on a phenomena related to the intestinal innate immune response of the orange- spotted grouper larvae affected by high temperatures. Fish were kept in three different temperatures: 28 0C (control), 28 0C slowly adjusted to 330C (acclimatized group), and 330C (shock group). After 7 days of acclimatizing corresponding temperatures, fish were injected with LPS, CpG ODN, and Poly I:C. Intestine samples were collected and divided to three parts: anterior gut; mid gut; and posterior gut. The mRNA was analyzed for gene expressions of related immune genes. Different TLRs and cytokines were tested. It is found that upon LPS stimulation, there different patterns on the expressions. Up- regulations and down- regulations were recorded as effect of high temperature in different sections of the intestine. Moreover, stimulation with poly I:C shows negligible effect of high temperature in different sections. Stimulation with ODN, on the other hand, has activated TLR 9A upon recognition with generally up- regulation or no adversary effect of high temperature on the expressions of immune related genes in different intestinal sections.
Acknowledgements i
摘要 ii
Abstract iii
List of Figures vi
Chapter 1 : Introduction 1
Chapter 2: Literature Review 4
2.1 Orange- spotted grouper (Epinephelus coioides) 4
2.1.1 Taxonomic Classification 4
2.1.2 Morphology 4
2.1.3 Habitat, Biology, and Distribution 4
2.1.4 Grouper Aquaculture 5
2.2 Fish Immunity 5
2.2.1 Fish Immune System 6
2.2.1.1 Intestinal Immunity 7
2.2.1.2 Innate Immunity 8
2.2.1.2.1 TLR 8
2.2.1.2.2 Interferon 10
2.2.1.2.3 IL- 1β 10
2.3 Immunostimulants 11
2.3.1. Poly I:C 5
2.3.2 LPS 11
2.3.3 CpG ODN 12
2.4 Environmental Factors 13
2.4.1 pH 13
2.4.2 Temperature 14
Chapter 3: Materials and Methods 15
3.1 Experimental Fish 15
3.2 Experimental Set- up 15
3.3 Immunostimulation and Sample Collection 15
3.4 RNA Extraction 16
3.5 cDNA Synthesis 17
3.6 qPCR 18
3.7 Statistical Analysis 19
Chapter 4: Results 20
4.1 Immunostimulation with LPS 20
4.2 Immunostimulation with Poly I:C 21
4.3 Immmunostimulation with CpG ODN 22
Chapter 5: Discussion 25
5.1 Immunostimulation with LPS 25
5.2 Immunostimulation with Poly I:C 26
5.3 Immmunostimulation with CpG ODN 28
Chapter 6: Conclusion30
Chapter 7: Figures 31
Chapter 8: References 44
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