[논문]Expression Analysis of Visual Arrestin gene during Ocular Development of Olive Flounder (Paralichthys olivaceus)

Yang, Hyun; Lee, Young Mee; Noh, Jae Koo; Kim, Hyun Chul; Park, Choul-Ji; Park, Jong-Won; Hwang, In Joon; Kim, Sung Yeon; Lee, Jeong-Ho

doi:10.12717/dr.2013.17.3.231

Expression Analysis of Visual Arrestin gene during Ocular Development of Olive Flounder (Paralichthys olivaceus) 원문보기

Development & reproduction = 발생과 생식, v.17 no.3, 2013년, pp.231 - 240

Yang, Hyun (Genetics and Breeding Research Center, NFRDI) , Lee, Young Mee (Genetics and Breeding Research Center, NFRDI) , Noh, Jae Koo (Genetics and Breeding Research Center, NFRDI) , Kim, Hyun Chul (Genetics and Breeding Research Center, NFRDI) , Park, Choul-Ji (Genetics and Breeding Research Center, NFRDI) , Park, Jong-Won (Genetics and Breeding Research Center, NFRDI) , Hwang, In Joon (Genetics and Breeding Research Center, NFRDI) , Kim, Sung Yeon (Genetics and Breeding Research Center, NFRDI) , Lee, Jeong-Ho (Genetics and Breeding Research Center, NFRDI)

Abstract ▼ AI-Helper

Olive flounder (Paralichthys olivaceus) is one of the commercial important flatfish species in Korea. The ocular signal transduction pathway is important in newly hatched flounders because it is closely involved in the initial feeding phase thus essential for survival during the juvenile period. However, the study of gene expression during ocular development is incomplete in olive flounder. Therefore we examined the expression analysis of specifically induced genes during the development of the visual system in newly hatched flounders. We searched ocular development-involved gene in the database of expressed sequence tags (ESTs) from olive flounder eye and this gene similar to arrestin with a partial sequence homology. Microscopic observation of retinal formation corresponded with the time of expression of the arrestin gene in the developmental stage. These results suggest that arrestin plays a vital role in the visual signal transduction pathway of the retina during ocular development. The expression of arrestin was strong in the ocular system during the entirety of the development stages. Our findings regarding arrestin have important implications with respect to its biological role and evolution of G-protein coupled receptor (GPCR) signaling in olive flounder. Further studies are required on the GPCR-mediated signaling pathway and to decipher the functional role of arrestin.

주제어

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문제 정의

However, the temporal expression patterns of the arrestin gene during the developmental stages and in other tissues of olive flounder remain unknown. The purpose of this study was to identify the genes expressed during ocular formation in the developmental stages of olive flounder.

제안 방법

Although expression of the arrestin gene was not detectable post-fertilization, induction of the arrestin was first detected from 1 DAH and continuously increased to 5 DAH and then gradually decreased in 6－7 DAH larvae. For further study of arrestin expression, a real time PCR assay was performed with olive flounder of various growth stage (Fig. 3A). Arrestin was similarly expressed in the eye and brain in 90-day-old flounder but not detected in other tissues.

대상 데이터

All experimental fish were raised at Genetics and Breeding Research Center, National Fisheries Research and Development Institute (NFRDI) and maintained in 10 tons flow through tank at 20±1℃ under a natural photoperiod.
The samples were examined under a stereo microscope (ZEISS CL1500 ECO Jena, Germany) imaging system at ×400 magnification of development was determined.

데이터처리

For unpaired matched comparative analysis of multiple groups, an analysis of variance (ANOVA) was performed. Data that did not meet normality assumptions were subjected to Kruskal-Wallis ANOVA on ranks, and then pair wise comparisons were made using the Student-Newman-Keuls (SNK) method.
Data were analyzed using Sigma plot for Windows (Jandel Scientific, San Rafael, CA, USA). For unpaired matched comparative analysis of multiple groups, an analysis of variance (ANOVA) was performed. Data that did not meet normality assumptions were subjected to Kruskal-Wallis ANOVA on ranks, and then pair wise comparisons were made using the Student-Newman-Keuls (SNK) method.

이론/모형

3%) respectively. The relative quantification of arrestin gene expression was performed using the comparative C_t method. The C_t value is defined as the point where a statistically significant increase in the fluorescence has occurred.
0 × e^－4 were considered to be significant, all ESTs were not identified as orthologs of known genes were designated as unknown EST clones. The trans-membrane secondary structure prediction program was used to TMRPres2D tool (Athens, Greece).

성능/효과

Experiments are examined with various periods of adult fish. (A) Arrestin gene expression from the segmental tissue (B: brain, M: muscle, F: fin, E: eye, L: liver, S: spleen, K: kidney, G: gill, N: negative) for the olive flounder at 90 days, 10 months and 2 years of age were analyzed using RT-PCR. (B) The mRNA expression of arrestin was analyzed by real-time PCR.
In conclusion, bioinformatics and expression analysis of visual arrestin during the stages of optical formation may assist the study of the initial larval life and growth in the olive flounder. Further functional studies on this gene will provide more information on molecular regulatory mechanisms, identity of arrestin subunit genes, and GPCR signaling pathways that are involved in ocular formation in the olive flounder.

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