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Phenotypic Assessment of Lipid Droplets using Daphnia magna Model
Kojo Eghan,이상우,김우근 환경독성보건학회 2021 한국독성학회 심포지움 및 학술발표회 Vol.2021 No.5
Lipids storage, accumulation and metabolism in organisms can be altered by several factors including the environment, exogenous and endogenous compounds. Additional energy stores in the form of lipids gives living organisms a substantial metabolic benefit, nonetheless, an energy balance disorder could lead to lipid diseases and liver dysfunctions. For crustaceans, this tends to affect their overall growth and development. In Daphnia magna, growth, molting, and reproductive functions are regulated by the amount and fate of storage lipids. Tributyltin (TBT), one of the well-known toxic pollutants to be found in the aquatic environment, is a substance that resists degradation and is absorbed by organic materials such as bacteria and algae. In this study, we seek to develop a model that can be used to study the mechanisms involved in lipid metabolism associated with accumulation, uptake and regulation. With little know about the formation and metabolism of lipid droplets in Daphnia manga both physically and transcriptionally, we employed this in vivo model in our study. Daphnia magna individuals of 5 days old were exposed to TBT 0, 1.0 and 1.0 μg/L for 48 h. Then, Oil Red O and Nile red staining processes were carried out, stained individuals were observed under a stereo microscope and confocal microscope, respectively. The transcription of selected genes (rxr, cer2, hr96, magro, man, NPC1b & SM3) involved in fat metabolism was also analyzed using qPCR techniques. From our findings, microscopic Daphnia magna images from both Nile red and Oil red staining showed analogous patterns. At the transcription level, rxr, cer2 and SM3 had being significantly altered. The Daphnia magna successfully responded to our tests and significant effects were observed as expected. Our findings indicate that TBT facilitates the uptake and accumulation of lipids in Daphnia magna. Therefore, the Daphnia magna model presents an endless potential to tap into, and can be an alternative to test lipid metabolism altering chemicals. Nonetheless, further research needs to be done on lipid metabolism mechanobiology, to understand how cells respond to mechanical signals that causes a disruption in lipids production and use.