Cyst viability and stress tolerance upon heat shock protein 70 knockdown in the brine shrimp Artemia franciscana
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Cyst viability and stress tolerance upon heat shock protein 70 knockdown in the brine shrimp Artemia franciscana Mat Taib Mimi Iryani 1 & Patrick Sorgeloos 1,2 & Muhd Danish-Daniel 1 & Min Pau Tan 1 & Li Lian Wong 1 & Wen Jye Mok 1 & Woro Hastuti Satyantini 3 & Gunanti Mahasri 3 & Yeong Yik Sung 1,3 Received: 2 January 2020 / Revised: 14 April 2020 / Accepted: 22 April 2020 # Cell Stress Society International 2020
Abstract Females of the brine shrimp Artemia franciscana produce either free-swimming nauplii via ovoviviparous pathway of reproduction or encysted embryos, known as cysts, via oviparous pathway, in which biological processes are arrested. While previous study has shown a crucial role of ATP-dependent molecular chaperone, heat shock protein 70 (Hsp70) in protecting A. franciscana nauplii against various abiotic and abiotic stressors, the function of this protein in diapausing embryos and cyst development, however, remains unknown. RNA interference (RNAi) was applied in this study to examine the role of Hsp70 in cyst development and stress tolerance, with the latter performed by desiccation and freezing, a common method used for diapause termination in Artemia cysts. Hsp70 knockdown was apparent in cysts released from females that were injected with Hsp70 dsRNA. The loss of Hsp70 affected neither the development nor morphology of the cysts. The time between fertilization and cyst release from Artemia females injected with Hsp70 dsRNA was delayed slightly, but the differences were not significant when compared to the controls. However, the hatching percentage of cysts which lacks Hsp70 were reduced following desiccation and freezing. Taken together, these results indicated that Hsp70 possibly plays a role in the stress tolerance but not in the development of diapause-destined embryos of Artemia. This research makes fundamental contributions to our understanding of the role molecular chaperone Hsp70 plays in Artemia, an excellent model organism for diapause studies of the crustaceans. Keywords Hsp70 . Artemia franciscana . Cysts . RNA interference (RNAi)
Introduction Brine shrimps Artemia are termed as “extremophiles” due to their ability to withstand a wide range of environmental stresses (Chen et al. 2007). Under favorable conditions, live nauplii emerge from the egg sac of Artemia females, which then undergo a series of molts before reaching adulthood (Liang and MacRae 1999). Alternatively, in anticipation of unfavorable
* Yeong Yik Sung [email protected] 1
Institute of Marine Biotechnology, Universiti Malaysia Terengganu (UMT), 21030 Kuala Nerus, Malaysia
2
Laboratory of Aquaculture and Artemia Reference Center, Department of Animal Sciences and Aquatic Ecology, Ghent University, Coupure Links 653, 9000 Ghent, Belgium
3
Department of Fish Health Management and Aquaculture, Faculty of Fisheries and Marine, Universitas Airlangga, Surabaya 60115, Indonesia
or extreme conditions, females release oviparous-developing encysted embryos which enter diapause. Once diapause is terminat
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