Knockdown of heat shock protein 70 (Hsp70) by RNAi reduces the tolerance of Artemia franciscana nauplii to heat and bacterial infection

Iryani, Mat Taib Mimi; MacRae, Thomas H.; Panchakshari, Sheethal; Tan, Jiabo; Bossier, Peter; Wahid, Mohd Effendy Abd; Sung, Yeong Yik

doi:10.1016/j.jembe.2016.12.004

Cited by 34 publications

(27 citation statements)

References 37 publications

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“…The dsRNA specific to hsp70 and gfp was mixed separately in 1:10 ratio (v/v) with 0.5% phenol red in Dulbecco’s phosphate-buffered saline (Sigma-Aldrich). The diluted dsRNA either hsp70 or gfp (250 nl of solution containing approximately 80 ng dsRNA) was injected with a FemtoJet ® microinjector (Eppendorf, USA) using Femtotips II microinjection capillary tips (Eppendorf, USA) to egg sacs of adult Artemia female while viewing under stereomicroscope (Figure S3 in Supplementary Material) ( 18 ). A total of thirty numbers of females were injected with dsRNA for each hsp70 and gfp .…”

Section: Methodsmentioning

confidence: 99%

“…), and collected in 1.5-ml sterile eppendorf tubes. The larvae were homogenized with a sterile microfuge pestle in 100 µl of TRIzol ® reagent (Invitrogen, USA) ( 15 , 18 ). The homogenate was centrifuged at 12,000 × g for 10 min at 4°C, and the supernatant was transferred to new sterile eppendorf tubes prior to RNA isolation according to the manufacturer’s protocols.…”

Section: Methodsmentioning

confidence: 99%

“…The brine shrimp ( Artemia franciscana ), an aquatic invertebrate that can be reared under gnotobiotic conditions (allowing full control over the host-associated microbial communities), is characteristically small, highly osmotolerant, branchiopod crustacean reported from a variety of harsh environment worldwide ( 14 ). Apart from its unusual life history, relatively low space and cost requirement for culture, rapid generation cycle (cyst to adult in 20–30 days), well-characterized developmental stages, the ability to form cyst that can be stored and used, and established molecular techniques like RNA interference (RNAi) make A. franciscana an exceptional experimental system to study the biological activity of polyphenol plant-based compounds as it allows to distinguish the direct effect on the host (by pre-exposing axenic brine shrimp larvae to the compound for a certain duration) from indirect effects ( 15 – 18 ).…”

Section: Introductionmentioning

confidence: 99%

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Phloroglucinol-Mediated Hsp70 Production in Crustaceans: Protection against Vibrio parahaemolyticus in Artemia franciscana and Macrobrachium rosenbergii

et al. 2018

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The halophilic aquatic bacterium, Vibrio parahaemolyticus, is an important aquatic pathogen, also capable of causing acute hepatopancreatic necrosis disease (AHPND) in shrimp resulting in significant economic losses. Therefore, there is an urgent need to develop anti-infective strategies to control AHPND. The gnotobiotic Artemia model is used to establish whether a phenolic compound phloroglucinol is effective against the AHPND strain V. parahaemolyticus MO904. We found that pretreatment with phloroglucinol, at an optimum concentration (30 µM), protects axenic brine shrimp larvae against V. parahaemolyticus infection and induced heat shock protein 70 (Hsp70) production (twofolds or more) as compared with the control. We further demonstrated that the Vibrio-protective effect of phloroglucinol was caused by its prooxidant effect and is linked to the induction of Hsp70. In addition, RNA interference confirms that phloroglucinol-induced Hsp70 mediates the survival of brine shrimp larvae against V. parahaemolyticus infection. The study was validated in xenic Artemia model and in a Macrobrachium rosenbergii system. Pretreatment of xenic brine shrimp larvae (30 µM) and Macrobrachium larvae (5 µM) with phloroglucinol increases the survival of xenic brine shrimp and Macrobrachium larvae against subsequent V. parahaemolyticus challenge. Taken together, our study provides substantial evidence that the prooxidant activity of phloroglucinol induces Hsp70 production protecting brine shrimp, A. franciscana, and freshwater shrimp, M. rosenbergii, against the AHPND V. parahaemolyticus strain MO904. Probably, phloroglucinol treatment might become part of a holistic strategy to control AHPND in shrimp.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Phloroglucinol-Mediated Hsp70 Production in Crustaceans: Protection against Vibrio parahaemolyticus in Artemia franciscana and Macrobrachium rosenbergii

et al. 2018

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“…Natural thermal tolerance in Artemia, and the capacity for its induction, varies across life stages and populations (Frankenberg et al, 2000;Clegg and Trotman, 2002); for example, populations from Vietnam display greater tolerance of high temperatures than those from San Francisco. Artemia thermal tolerance across populations is probably regulated in part by heat shock protein 70 (HSP70) (Iryani et al, 2017). Artemia have been used in several previous heat-hardening experiments (Miller and McLennan, 1988;Frankenberg et al, 2000;Norouzitallab et al, 2014); induced thermal tolerance in these examples is achieved via repeated exposure to sublethal heat shocks varying from 40 to 50°C.…”

Section: Introductionmentioning

confidence: 99%

Can physiological engineering/programming increase multi-generational thermal tolerance to extreme temperature events?

Sorby

Green

Dempster

et al. 2018

Journal of Experimental Biology

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Organisms increasingly encounter higher frequencies of extreme weather events as a consequence of global climate change. Currently, few strategies are available to mitigate climate change effects on animals arising from acute extreme high-temperature events. We tested the capacity of physiological engineering to influence the intra- and multi-generational upper thermal tolerance capacity of a model organism, , subjected to extreme high temperatures. Enhancement of specific physiological regulators during development could affect thermal tolerance or life-history attributes affecting subsequent fitness. Using experimental populations, we exposed F0 individuals to one of four treatments: heat hardening (28°C to 36°C, 1°C per 10 min), heat hardening plus serotonin (0.056 μg ml), heat hardening plus methionine (0.79 mg ml) and a control treatment. Regulator concentrations were based on previous literature. Serotonin may promote thermal tolerance, acting upon metabolism and life history. Methionine acts as a methylation agent across generations. For all groups, measurements were collected for three performance traits of individual thermal tolerance (upper sublethal thermal limit, lethal limit and dysregulation range) over two generations. The results showed that no treatment increased the upper thermal limit during acute thermal stress, although serotonin-treated and methionine-treated individuals outperformed controls across multiple thermal performance traits. Additionally, some effects were evident across generations. Together, these results suggest that phenotypic engineering provides complex outcomes, and if implemented with heat hardening can further influence performance in multiple thermal tolerance traits, within and across generations. Potentially, such techniques could be up-scaled to provide resilience and stability in populations susceptible to extreme temperature events.

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“…Survival of A. franciscana cysts depends on several structural and molecular adaptations including the presence of molecular chaperones (MacRae, 2016), of which the ATP-independent small heat shock proteins (sHsps) have received the most attention (Jackson and Clegg, 1996;Liang and MacRae, 1999;Liang et al, 1997a,b;King and MacRae, 2012;King et al, 2013). ATPdependent molecular chaperones have also been examined in A. franciscana and these include Hsp70 (Clegg, 2011;Iryani et al, 2017), a major heat shock protein (Hsp) responsible for the folding of nascent proteins, promoting the assembly of protein complexes and either salvaging or destroying aberrant proteins (Mayer and Bukau, 2005;Zuiderweg et al, 2017;Zẇirowski et al, 2017;Mayer, 2018). These activities depend on conformational changes in Hsp70 modulated by its ATPase activity and this is influenced by co-chaperones such as the J-domain proteins or Hsp40s (Mayer and Bukau, 2005;Craig and Marszalek, 2017;Bascos et al, 2017;Kityk et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

ArHsp40 and ArHsp40-2 contribute to stress tolerance and longevity in Artemia franciscana, but only ArHsp40 influences diapause entry

Rowarth

MacRae

2018

Journal of Experimental Biology

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Embryos of the crustacean develop either ovoviviparously or oviparously, yielding swimming larvae (nauplii) or encysted gastrulae (cysts), respectively. Nauplii moult several times and become adults whereas cysts enter diapause, a state of dormancy characterized by exceptionally low metabolism and high stress tolerance. Synthesis of molecular chaperones such as the J-domain proteins ArHsp40 and ArHsp40-2 occurs during embryo development and post-diapause growth of and they influence development and stress tolerance. To further investigate J-domain protein function, ArHsp40 and ArHsp40-2 were each knocked down by RNA interference. Reductions in ArHsp40 and ArHsp40-2 had no effect on adult survival, time to release of cysts and nauplii from females and first-brood size. However, knockdown of both J-domain proteins reduced the longevity and heat tolerance of nauplii, with the loss of ArHsp40 having a greater effect. The knockdown of ArHsp40, but not of ArHsp40-2, caused approximately 50% of cysts to abort diapause entry and hatch without exposure to an exogenous signal such as low temperature and/or desiccation. Cysts lacking ArHsp40 that entered diapause exhibited decreased stress tolerance as did cysts with reduced ArHsp40-2, the latter to a lesser degree. The longevity of nauplii hatching prematurely from cysts was less than for nauplii arising by other means. The results expand our understanding of Hsp40 function in stress tolerance and development, especially during diapause, and they provide the first example of a molecular chaperone that influences diapause entry.

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Knockdown of heat shock protein 70 (Hsp70) by RNAi reduces the tolerance of Artemia franciscana nauplii to heat and bacterial infection

Cited by 34 publications

References 37 publications

Phloroglucinol-Mediated Hsp70 Production in Crustaceans: Protection against Vibrio parahaemolyticus in Artemia franciscana and Macrobrachium rosenbergii

Phloroglucinol-Mediated Hsp70 Production in Crustaceans: Protection against Vibrio parahaemolyticus in Artemia franciscana and Macrobrachium rosenbergii

Can physiological engineering/programming increase multi-generational thermal tolerance to extreme temperature events?

ArHsp40 and ArHsp40-2 contribute to stress tolerance and longevity in Artemia franciscana, but only ArHsp40 influences diapause entry

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