활성산소종(ROS)은 환경 스트레스 및 병원체의 침입에 의한 산화 대사의 자연적인 산물로써 생물에서 생산된다. 산화적 스트레스에 의해 생성되는 superoxide 음이온 및 과산화수소와 같은 ROS는 세포와 조직에 독성을 나타낼 수 있고, 이 과정에 관여하는 superoxide dismutase (SOD)는 중요한 metalloenzyme이다. 최근 연구는 올리브 넙치(Paralichthys olivaceus)에서 SOD의 부분 유전자가 benzo[a]pyrene에 의해 강하게 발현이 유도되고 산화 스트레스 반응의 지표라고 확인하였지만, 바이러스성 감염에 대한 전사적 반응에 대해서는 조사되지 않았다. 본 연구에서는 항바이러스 반응에서 넙치 SOD의 기능을 알아보기 위해 공간 및 시간적 발현 프로파일을 분석하였다. 넙치 SOD 전사체는 정도의 차이는 있지만 다양한 기관에서 보편적으로 발현되었으며, 근육, 간, 뇌에서는 높게 발현되었고, 위와 비장에서는 상대적으로 낮게 발현되었다. VHSV 감염 후 넙치 콩팥에서 SOD 발현은 3시간 이내에 증가하였으며 점차적으로 감소하여 감염 2일째 원래 수준으로 돌아갔다. 검사 조직에 따라 발현이 유도되는 시간의 차이는 있지만 근육, 간, 뇌에서도 콩팥과 유사한 발현양상을 보였으며, 공통적으로 급성적 면역반응에서는 발현이 증가하지만 만성적 면역반응에서는 감소하였다. 이상의 결과들을 종합해 볼 때, 넙치 SOD는 넙치(P. olivaceus)의 면역 방어 시스템에 중요한 역할을 하고 넙치의 산화 스트레스에 대한 보호 효과에 기여할 것으로 기대할 수 있다.
활성산소종(ROS)은 환경 스트레스 및 병원체의 침입에 의한 산화 대사의 자연적인 산물로써 생물에서 생산된다. 산화적 스트레스에 의해 생성되는 superoxide 음이온 및 과산화수소와 같은 ROS는 세포와 조직에 독성을 나타낼 수 있고, 이 과정에 관여하는 superoxide dismutase (SOD)는 중요한 metalloenzyme이다. 최근 연구는 올리브 넙치(Paralichthys olivaceus)에서 SOD의 부분 유전자가 benzo[a]pyrene에 의해 강하게 발현이 유도되고 산화 스트레스 반응의 지표라고 확인하였지만, 바이러스성 감염에 대한 전사적 반응에 대해서는 조사되지 않았다. 본 연구에서는 항바이러스 반응에서 넙치 SOD의 기능을 알아보기 위해 공간 및 시간적 발현 프로파일을 분석하였다. 넙치 SOD 전사체는 정도의 차이는 있지만 다양한 기관에서 보편적으로 발현되었으며, 근육, 간, 뇌에서는 높게 발현되었고, 위와 비장에서는 상대적으로 낮게 발현되었다. VHSV 감염 후 넙치 콩팥에서 SOD 발현은 3시간 이내에 증가하였으며 점차적으로 감소하여 감염 2일째 원래 수준으로 돌아갔다. 검사 조직에 따라 발현이 유도되는 시간의 차이는 있지만 근육, 간, 뇌에서도 콩팥과 유사한 발현양상을 보였으며, 공통적으로 급성적 면역반응에서는 발현이 증가하지만 만성적 면역반응에서는 감소하였다. 이상의 결과들을 종합해 볼 때, 넙치 SOD는 넙치(P. olivaceus)의 면역 방어 시스템에 중요한 역할을 하고 넙치의 산화 스트레스에 대한 보호 효과에 기여할 것으로 기대할 수 있다.
Superoxide dismutase is a family of important antioxidant metalloenzymes and catalyzes the dismutation of toxic superoxide anions into dioxygen and hydrogen peroxide. A recent study identified the partial superoxide dismutase (SOD) gene in olive flounder (Paralichthys olivaceus). The same study repo...
Superoxide dismutase is a family of important antioxidant metalloenzymes and catalyzes the dismutation of toxic superoxide anions into dioxygen and hydrogen peroxide. A recent study identified the partial superoxide dismutase (SOD) gene in olive flounder (Paralichthys olivaceus). The same study reported that it strongly induced benzo[a]pyrene and that it was an indicator of aquatic oxidative stress responses. However, its transcriptional response against viral infection has not been investigated. In the present study, the spatial and temporal expression profiles were analyzed to investigate the function of Of-SOD in the antiviral response. The Of-SOD transcripts were ubiquitously detected at various levels in diverse tissues in a real-time PCR. The expression of Of-SOD was significantly higher in the muscles, liver, and brain but extremely low in the stomach and spleen. Following a VHSV challenge, the expression of Of-SOD increased within 3 h in the kidneys and decreased to the original level 2 days postchallenge. In muscle, liver, and brain, Of-SOD mRNA was similarly up-regulated at 3-6 h postchallenge and then decreased to the basal level. Although the expression pattern and induction time differed slightly depending on the tissue, the transcript of Of-SOD consistently increased in the acute infection response, but the expression was low in the chronic response. The expression of Of-SOD was induced after the VHSV infection, and Of-SOD was probably involved in the immune response against the viral challenge. These results suggest that SOD may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in olive flounder.
Superoxide dismutase is a family of important antioxidant metalloenzymes and catalyzes the dismutation of toxic superoxide anions into dioxygen and hydrogen peroxide. A recent study identified the partial superoxide dismutase (SOD) gene in olive flounder (Paralichthys olivaceus). The same study reported that it strongly induced benzo[a]pyrene and that it was an indicator of aquatic oxidative stress responses. However, its transcriptional response against viral infection has not been investigated. In the present study, the spatial and temporal expression profiles were analyzed to investigate the function of Of-SOD in the antiviral response. The Of-SOD transcripts were ubiquitously detected at various levels in diverse tissues in a real-time PCR. The expression of Of-SOD was significantly higher in the muscles, liver, and brain but extremely low in the stomach and spleen. Following a VHSV challenge, the expression of Of-SOD increased within 3 h in the kidneys and decreased to the original level 2 days postchallenge. In muscle, liver, and brain, Of-SOD mRNA was similarly up-regulated at 3-6 h postchallenge and then decreased to the basal level. Although the expression pattern and induction time differed slightly depending on the tissue, the transcript of Of-SOD consistently increased in the acute infection response, but the expression was low in the chronic response. The expression of Of-SOD was induced after the VHSV infection, and Of-SOD was probably involved in the immune response against the viral challenge. These results suggest that SOD may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in olive flounder.
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문제 정의
Regarding the problem, a better understanding of its immune response should help to reduce diseases in addition to immune-related genes. In this study, we demonstrated the spatial and temporal expression of Of-SOD from olive flounder in order to investigate the distribution and functional roles of Of-SOD against viral infection.
제안 방법
In this study, we analyzed the expression of Of-SOD spatial and temporal expression to investigate a role of Of-SOD against the invasion of pathogens. The results indicated that Of-SOD transcripts were constitutively expressed in all examined tissues, but the relative levels of expression were variable.
After challenge, fish were transferred to new 3 ton tank and 5% of the water was exchanged everyday. Samples were collected at 0, 1, 3, 6, 9, 12, 18, 24 hr and 2, 3, 4, 5, 6, 7, 10, 15, 20 days post-injection to collect above the described tissues and total RNA was extracted from the tissues and cDNA was synthesized as described.
Analyzed tissue is: brain, eye, fin, gill, intestine, kidney, liver, muscle, spleen and stomach. This experiment was performed for three times and the data demonstrated means of triplicates. Error bars in dicate SD.
To investigate the function of Of-SOD in the viral-infection response, the olive flounder fish were challenged with VHSV and the kidneys dissected out in a variety of infection time of the fish owing to its importance in the immune system, and the expression of Of-SOD was assayed by real-time PCR. The expression of Of-SOD was increased a 2.
To investigate the tissue distribution profile of Of-SOD transcripts, total RNA from the tissues of brain, eye, fin, gill, intestine, kidney, liver, spleen and stomach was excised from healthy olive flounder. The real-time PCR analysis was performed using olive flounder β-actin as an invariant control and relative mRNA expression-fold was derived by comparing the transcript level in each tissue with that of stomach.
성능/효과
The real-time PCR analysis was performed using olive flounder β-actin as an invariant control and relative mRNA expression-fold was derived by comparing the transcript level in each tissue with that of stomach. As a result, SOD transcripts were detected in all the examined tissues, but the relative levels of basal expression were variable. Of the ten tissues examined, the highest level of expression was observed in the muscle, followed by liver, brain.
In this study, we analyzed the expression of Of-SOD spatial and temporal expression to investigate a role of Of-SOD against the invasion of pathogens. The results indicated that Of-SOD transcripts were constitutively expressed in all examined tissues, but the relative levels of expression were variable. Predominant expression of Of-SOD was detected in muscle, liver and brain, while relatively low in spleen and stomach.
참고문헌 (21)
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