[논문]Expression of Perforin Gene for Early Development of Nephrons in Olive Flounder (Paralichthys olivaceus)

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

doi:10.12717/dr.2013.17.4.321

Expression of Perforin Gene for Early Development of Nephrons in Olive Flounder (Paralichthys olivaceus) 원문보기

Development & reproduction = 발생과 생식, v.17 no.4, 2013년, pp.321 - 327

Yang, Hyun (Genetics and Breeding Research Center, NFRDI) , Lee, Young Mee (Genetics and Breeding Research Center, NFRDI) , Lee, Jeong-Ho (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)

Abstract ▼ AI-Helper

The innate immune system is the only defense weapon that invertebrates have, and it is the fundamental defense mechanism for fish. The innate immune response is important in newly hatched flounders because it is closely involved in the initial feeding phase, which is why it is essential for survival during the juvenile period. The expression analysis of genes involved in the innate immune response in the olive flounder (Paralichthys olivaceus) in the days after hatching is incomplete. Therefore, we have begun to examine the expression patterns of genes specifically induced during the development of the innate immune system in newly hatched flounders. Microscopic observation showed that pronephron formation corresponded with the expression of perforin-encoding gene. These results suggest that perforin plays a vital role in the innate immunity of the kidney during developmental stages. Perforin expression was strong at the start of the development of the innate immune response, and continued throughout all the development stages. Our findings have important implications with respect to perforin's biological role and the evolution of the first defense mechanisms in olive flounder. Further studies are required to elucidate the perforin-mediated innate immunity response and to decipher the functional role of perforin in developmental stages.

주제어

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문제 정의

However, the temporal expression patterns of this perforin-like gene during the developmental stages and in normal tissues of olive flounder are unknown. The purpose of this study was to examine the expression patterns of perforin in normal tissues of the olive flounder and during the development of the innate immune system in newly hatched olive flounders.

제안 방법

However, the formation of the pronephric tubules and glomeruli has not been described in any study. Guided by the position of the mature pronephric nephron, we examined the cellular structures medial to the anterior tips of the pronephric ducts at various time points to define the stages of nephron formation. At 1-DAH, a nephron primordium was visualized ventral to the third somite (Fig.

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 deter- mined.

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 one way 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 one way ANOVA on ranks, and then pair wise comparisons were made using the Student-Newman-Keuls (SNK) method.

이론/모형

Embryo and larva were harvested during hours post fertilization (HPF) and days after hatching (DAH) for the indicated periods. The RNA extracted and perforin was analyzed by RT-PCR method. (B) The mRNA ex- pression of perforin was analyzed by real-time PCR.
4%) respectively. The relative quantification of perforin 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.

성능/효과

Experiments are examined with various periods of adult fish. (A) Perforin gene ex- pression from the segmental tissue (B: brain, M: muscle, F: fin, E: eye, L: liver, S: spleen, K: kidney, G: gill) for the olive flounder at 2 years of age were analyzed using RT-PCR. (B) The mRNA expression of perforin was analyzed by real-time PCR.
In conclusion, expression analysis of cytotoxic perforin during pronephron formation might provide information that could be useful in understanding the initial immune responses and growth of olive flounder larvae. Further functional studies on this gene will help elucidate the molecular regulatory mechanisms and the immune signaling pathways involved in pronephron formation in olive flounder, as well as identifying the perforin-mediated genes.

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