RNA interference of Hsp70 in Artemia franciscana nauplii and its effect on morphology, growth, survival and immune response

Iryani, Mat Taib Mimi; MacRae, Thomas H.; Sorgeloos, Patrick; Muhammad, Tengku Sifzizul Tengku; Danish-Daniel, Muhd; Tan, Min Pau; Satyantini, Woro Hastuti; Wahid, Mohd Effendy Abd; Sun, Jingfeng; Lv, Aijun; Sung, Yeong Yik

doi:10.1016/j.aquaculture.2020.735012

Cited by 12 publications

(4 citation statements)

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“…Knockdown of Hsp70 with this method was shown to be transient, as the protein which was initially undetected in nauplii stage was expressed in juvenile and adult stages of Artemia. The Hsp70-knockdown nauplii developed normally, with their morphology similar to those accumulating normal amounts of the protein (Iryani et al 2020). Here, we continue this study to unravel the function of Hsp70 in A. franciscana during diapause with the aim to first demonstrate the presence of Hsp70 in the cysts.…”

Section: Introductionmentioning

confidence: 75%

“…Forward (5′-TAATACGACTCACTATAGGGATTCTCAAAGAC AAGC-3′) and reverse (5′-TAATAC GACTCACTATAGGG CATAGAGCTTGGTAAT-3′) primers, specific for Hsp70, were used for cDNA amplification by PCR using ImmoMix™ (Bioline, UK) at the manufacturer's suggested concentration. The PCR product was used as template to generate Hsp70 dsRNA using MEGAscript® RNAi kit (Ambion Applied Biosystems, USA) and green fluorescence protein (GFP) dsRNA was used as control solution (Iryani et al 2017;Iryani et al 2020).…”

Section: Preparation Of Hsp70 Dsrnamentioning

confidence: 99%

“…Males were introduced to females 24 h after injection. Mating pairs were fed daily with microalgae Chlorella vulgaris and females were monitored for fertilization (Iryani et al 2017;Iryani et al 2020). The time from fertilization to release of cysts from females was determined.…”

Section: Injection Of a Franciscana Females With Dsrna And Observation Of Embryo Developmentmentioning

confidence: 99%

“…Fifteen microliters of each sample was applied per lane of 10% SDS gels prior to electrophoresis and then blotted to PVDF membrane (BioRadImmun-Blot™ PVDF, USA) for antibody probing. PVDF membrane was probed in monoclonal antibody that recognize Hsp70 (SMC-164D) (StressMarq, Canada), followed by polyclonal HRP-conjugated goat anti-mouse IgG (Bioreagent-SAB-100J) (Stressgen, Canada) prior to the detection of antibody-reactive proteins (Iryani et al 2020).…”

Section: Detection Of Hsp70 Mrna and Protein In A Franciscana Cystsmentioning

confidence: 99%

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Cyst viability and stress tolerance upon heat shock protein 70 knockdown in the brine shrimp Artemia franciscana

Iryani

Sorgeloos

Danish-Daniel

et al. 2020

Cell Stress and Chaperones

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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.

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Section: Introductionmentioning

confidence: 75%

Section: Preparation Of Hsp70 Dsrnamentioning

confidence: 99%

Section: Injection Of a Franciscana Females With Dsrna And Observation Of Embryo Developmentmentioning

confidence: 99%

Section: Detection Of Hsp70 Mrna and Protein In A Franciscana Cystsmentioning

confidence: 99%

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