UV and O3 are materials used in the water treatment process, and many studies have been reported to remove organic matters, contaminants, and microorganisms. In this study, we were investigated effects of Chirnomidae (Chironomus flaviplumus, Chironomus riparius), which are contamination indicator species to exposure UV and O3 for the survival rate, body color change and gene expression response. The survival rate of C. flaviplumus exposed to UV decreased to about 70% after 24 hours, and C. riparius about 50%. There was no change in the survival rate of C. flaviplumus exposed to O3, and C. riparius decreased to 95% after 10 minutes of exposure, but there was no change during the subsequent exposure time. In addition, UV and O3 exposure to the two species in body color faded in a time-dependent. In the HSP70 gene expression, C. riparius showed an increase in expression after UV exposure compared to the control group, and a significant difference was shown 12 hours after exposure (P<0.05). C. flaviplumus exposed to O3 showed a relatively low expression compared to the control group, and showed a significant difference at 10 minutes and 1 hour after exposure (P<0.05). These results reported the ecotoxicological effects on Chironomidae according to UV and O3 exposure. Therefore, the results of this study can be used as basic data to understand the effects of UV and O3, which are disinfectants used in water treatment plants, on Chirnomidae entering plants. Key words: Chironomus flaviplumus, Chironomus riparius, UV, O3, acute toxicity, survival
돌기해삼 Apostichopus japonicus는 주요 양식 대상 무척추동물로서 우리나라 연안 해역에 서 식하고 있다. 본 연구는 방류 방법에 따른 단기간의 생리학적 스트레스 정도를 평가하기 위하 여 heat shock protein 90 (HSP90) 유전자의 발현 변화를 실시간 정량적 중합효소연쇄반응법 으로 조사하였다. 어린 돌기해삼을 비닐봉지에 산소 포장하여 30분간 수송하거나 방류 해역의 간조기에 1시간 공기 중에 노출된 실험군의 HSP90 유전자 발현은 대조군의 HSP90 유전자 발현에 비하여 통계학적으로 유의미하게 증가하였다(수송 후 실험군 p=0.001; 간조기 실험군 p=0.032). 어린 돌기해삼을 방류 후 6시간까지 분석한 결과, 선상에서 씨뿌림 방식으로 방류된 6시간째의 개체 및 호스를 통과하여 수중으로 방류된 2~6시간째의 HSP90 유전자 발현율은 대 조군에 비하여 약간 감소하는 경향을 보였다(씨뿌림 실험군 p=0.069; 호스 방류군 p=0.093). 한 편, 잠수부에 의해 수중에서 방류된 어린 돌기해삼은 방류 후 시간이 경과할수록 HSP90 유전 자 발현율은 증가하는 패턴이 관찰되었다(p=0.061). 이상의 결과는 방류된 어린 돌기해삼의 단기간 스트레스 반응 연구와 효과적인 방류 방법의 개발에 HSP90 유전자 발현이 유용하게 사용될 수 있음을 시사한다.
참다슬기 아가미 조직으로부터 heat shock protein 70 유전자를 분리 · 동정하였다. 참다슬기 HSP70 cDNA의 open reading frame (ORF)는 1,917 bp로 639개의 아미노산을 암호화하여 분자 량은 약 70 kDa으로 예측되었다. 생물정보학 배열분석에 의해 HSP 유전자 기능과 관여되어 있는 3가지 주요 signature motifs와 보존된 도메인을 확인하였다. 계통학적 분석을 통하여 참 다슬기 HSP70 유전자는 왕우렁이 Pomacea canaliculate와 같은 클러스트에 포함된다는 사실을 확인하였다. 수온 및 염분 변화에 따라, 참다슬기 HSP70 mRNA 유전자 레벨은 유의적으로 증 가하였으며(p < 0.05), 이는 외부자극요인을 파악할 있는 분자생물학적 마커로서 활용될 수 있 을 것으로 사료된다.
In order to examine if arsenic, one of environmental stresses, contributes to hypertension as one of cardiovascular pathological factors, this study was performed in vivo and in vitro, using intacted or pithed rats and aorta ring preparation, respectively. And also the relationship between expression of heat shock protein (HSP) 90 and vasoactives-induced contractile response was elucidated.
To measure blood pressure, the carotid arterial pressure was recorded on physiograph(Grass Co. 79E) connected to strain gauge. On the other hand, contractile response of vascular ring preparation isolated from rat was determined in organ bath and was recorded on physiograph connected to isometric transducer. And HSP was detacted by Western blotting whole cell lysis.
Preganglionic nerve stimulation was increased by 26.0% in arterial pressure of rat treated with arsenic.
Vascular contractile response was monitored and HSP were measured by Western blotting of whole lysis prepared from samples exposed with 0, 0.5, 1, 2 and 4 mM of arsenic for 8 hours. The dose-vascular responses of potassium chloride were augmented by increasing dose of arsenic in the strips exposed to arsenic for 8 hours, and were not augmented for 1, 3, 5 hours. And the response of relaxation of rat aorta induced by histamine was not influenced by arsenic stress.
The increase of HSP 90 expression in rat aorta was pronounced at 8 hours after 4 mM of arsenic treatment, but HSP 60 expression was not.
Arsenic stress not only increased the expression of HSP 90 in the rat aorta, but also augmented contractions to potassium chloride.
These results indicated that arsenic stress was sufficient to induce heat shock protein 90, resulting in increased vascular contractility in rat aorta.