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붉바리(Epinephelus akaara) 초기 성장 단계에 따른 망막의 발달과 opsin 유전자의 발현 양상

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Alternative Title
Retinal development and opsin gene expression during the juvenile development in red spotted grouper(Epinephelus akaara)
Abstract
In fish, photoreceptors including rods and cones are the membrane receptors that receive the light and transmit the light-induced signals to the brain regions to make an image of the outside world. Rod photorecetpors recognize the light and darkness, while cone photoreceptors respond to different wavelengths of light depending on their subtypes. To produce healthy and stable seed production, we need to obtain information and understand vision that affects behavior of red spotted grouper. We cloned their opsin genes here and examined their expression during the juvenile development. Using TA cloning method, we successfully partial cloned SWS2 (Short wavelength sensitive opsin), MWS (middle wavelength sensitive opsin), LWL (long wavelength sensitive opsin) and rod opsin in this fish species. Then, their tissue expression levels were determined by RT-PCR in various adult tissues including forebrain, midbrain, hindbrain, retina, kidney, heart, intestine, muscle, gonad and gill. We found that all opsin genes were strongly expressed in the retina. To examine their expression during the eye development, we isolated tissue samples at different developmental stages (lens and ear vesicle formation stage, 1, 2, 3, 5, 10, 21, 50 days after hatched) and determined their expression level by real-time quantitative PCR. SWS2, a cone photoreceptor, began to be expressed from lens and ear vesicle formation stage and its expression gradually increased until 10 dah. In case of MWS, its expression was detected 3 dah and reached the highest level at 21 dah. The expression of LWS was first observed from 3 dah and their expression decreased thereafter. Rhodopsin, a rod photoreceptor, was found to be expressed from 2 dah and its expression reached the highest level at 50 dah. The outer nuclear layer (ONL), inner nuclear layer (INL) and ganglion cell layer began to differentiate at 2 dah, while choroid first appeared at 4 dah so that the eyes became black. These results indicate that the development of retina mostly completes around 4 dah. It seems that the development of the retina and the expression of the opsin genes are closely related to the behavior such as hunting prey, considering that the timing of the completion of the development of the retina, the timing of gene expression, and the timing of completion of yolk absorption are similar.
Author(s)
김은수
Issued Date
2017
Awarded Date
2017. 8
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000008232
Alternative Author(s)
Kim, Eun Su
Affiliation
제주대학교 일반대학원
Department
대학원 해양생명과학과
Advisor
이영돈
Table Of Contents
목차 Ⅰ
List of Figure Ⅲ
List of Table Ⅴ
Abstract 1
Ⅰ. 서론 3
Ⅱ. 재료 및 방법
1. 실험어 6
2. 붉바리 초기 성장 단계에 따른 망막의 발달
1) 붉바리 자치어의 형태 발달 및 행동 특성 7
2) 붉바리 자치어 망막의 발달 7
3. Opsin 유전자의 발현 양상
1) Opsin 유전자 partial cloning 8
2) Opsin 유전자의 조직발현 및 성장단계별 발현 양상 분석을 위한 Total RNA 추출 및 cDNA 합성 10
3) Opsin 유전자의 발현 양상 분석을 위한 Real-time quantitative PCR 14
4. 통계처리 14
Ⅲ. 결과
1. 붉바리 자치어의 형태 발달과 행동특성15
2. 붉바리 자치어 망막의 구조와 발달 18
3. Opsin 유전자의 partial cloning 21
4. Opsin 유전자의 조직별 발현 30
5. 성장단계에 따른 Opsin 유전자의 발현 양상 33
Ⅳ. 고찰 39
참고문헌 42
Degree
Master
Publisher
제주대학교 일반대학원
Citation
김은수. (2017). 붉바리(Epinephelus akaara) 초기 성장 단계에 따른 망막의 발달과 opsin 유전자의 발현 양상
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General Graduate School > Marine Life Sciences
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