| 1. |
- Noonin, Chadanat, et al.
(författare)
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Circadian regulation of melanization and prokineticin homologues is conserved in the brain of freshwater crayfish and zebrafish
- 2013
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Ingår i: Developmental and Comparative Immunology. - : Elsevier BV. - 0145-305X .- 1879-0089. ; 40:2, s. 218-226
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Tidskriftsartikel (refereegranskat)abstract
- Circadian clock is important to living organisms to adjust to the external environment. This clock has been extensively studied in mammals, and prokineticin 2 (Prok2) acts as one of the messenger between the central nervous system and peripheral tissues. In this study, expression profiles of Prok1 and Prok2 were investigated in a non-mammalian vertebrate brain, zebrafish, and the expression was compared to the Prok homologues, astakines (Ast1 and Ast2) in crayfish. These transcripts exhibited circadian oscillation in the brain, and Ast1 had similar pattern to Prok2. In addition, the expression of tyrosinase, an enzyme which expression is regulated by E-box elements like in Prok2, was also examined in zebrafish brain and was compared with the expression of prophenoloxidase (proPO), the melanization enzyme, in crayfish brain. Interestingly, the expressions of both Tyr and proPO displayed circadian rhythm in a similar pattern to Prok2 and Ast1, respectively. Therefore, this study shows that circadian oscillation of prokineticin homologues and enzymes involved in melanization are conserved.
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| 2. |
- Söderhäll, Kenneth, et al.
(författare)
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DSCAM (Down syndrome adhesion molecule); structure and function in a crustacean.
- 2012
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- In invertebrates, the circulating blood cells (hemocytes) are crucial for protecting the animal against invading microorganisms, and they are directly involved in recognition, phagocytosis, melanization and cytotoxic reactions. Many microorganisms trigger hemocyte synthesis and release from the HPT, and this may be one way for the host to respond to an infection. Therefore we have studied hematopoiesis intensely and have now in detail deciphered the molecular mechanisms in hematopoietic stem cell proliferation and differentiation. Recently we have cloned Dscam proteins from P. leniusculus. The Down syndrome cell adhesion molecule (Dscam) is a member of the immunoglobulin super family of proteins. In vertebrates Dscam was first identified for its importance in developing neural tissue, and for its critical role in Down Syndrome.These cell adhesion molecules are large proteins containing ten Ig domains and six fibronectin type III domains, and this structure is conserved across animal phyla. There are two Dscam genes in humans (DSCAM), and there are evidences for a conserved essential role in neural wiring, in both vertebrate and insects. An intriguing difference in Dscam gene structure is the exon duplication that have occurred in arthropods and which give rise to an array of alternative spliced isoforms in insects and crustaceans. Alternative splicing of individual exon sequences occurs in a mutually exclusive manner and allows for expression of several thousands of isoforms.We have now shown that hemocytes that express different spliced isoforms are induced by specific challenge, and moreover that these specific forms are important in clearing bacteria. This differential splicing may be one new area of research that may help to explain how crustaceans and insects as effectively can manage to distinguish between harmless and harmful microorganisms.
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| 3. |
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| 4. |
- Watthanasurorot, Apiruck, et al.
(författare)
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A gC1qR Prevents White Spot Syndrome Virus Replication in the Freshwater Crayfish Pacifastacus leniusculus
- 2010
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Ingår i: Journal of Virology. - : American Society for Microbiology. - 0022-538X .- 1098-5514. ; 84:20, s. 10844-10851
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Tidskriftsartikel (refereegranskat)abstract
- The gC1qR/p32 protein is a multiple receptor for several proteins and pathogens. We cloned a gC1qR homologue in a crustacean, Pacifastacus leniusculus, and analyzed the expression of P. leniusculus C1qR (PlgC1qR) in various tissues. The gC1qR/p32 transcript was significantly enhanced by white spot syndrome virus (WSSV) infection 6 h after viral infection both in vitro in a hematopoietic tissue cell culture (Hpt) and in vivo compared to appropriate controls. Moreover, PlgC1qR silencing in both the Hpt cell culture and live crayfish enhanced the WSSV replication. In addition, by making a recombinant PlgC1qR protein we could show that if this recombinant protein was injected in a crayfish, Pacifastacus leniusculus, followed by injection of WSSV, this significantly reduced viral replication in vivo. Furthermore, if the recombinant PlgC1qR was incubated with Hpt cells and then WSSV was added, this also reduced viral replication. These experiments clearly demonstrate that recombinant PlgC1qR reduce WSSV replication both in vivo and in vitro. The results from a far-Western overlay and glutathione S-transferase pull-down assays showed that PlgC1qR could bind to VP15, VP26, and VP28. Altogether, these results demonstrate a role for PlgC1qR in antiviral activity against WSSV.
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| 5. |
- Watthanasurorot, Apiruck, et al.
(författare)
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A mammalian like interleukin-1 receptor-associated kinase 4 (IRAK-4), a TIR signaling mediator in intestinal innate immunity of black tiger shrimp (Penaeus monodon)
- 2012
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Ingår i: Biochemical and Biophysical Research Communications - BBRC. - : Elsevier BV. - 0006-291X .- 1090-2104. ; 417:1, s. 623-629
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Tidskriftsartikel (refereegranskat)abstract
- Interleukin-1 receptor associated kinase-4 (IRAK-4) has been identified as a central signal transduction mediator of the Toll-like receptor (TLR) and Toll/interleukin-1 receptor (TIR) pathways in vertebrate innate immunity. An IRAK-4 homologue was cloned from the black tiger shrimp (Penaeus monodon) (PmIRAK-4) and it shares domains and structures with other IRAK-4s. It was found to be mainly expressed in the hemocytes and midgut but also to a lower extent in several other tissues in shrimp. The PmIRAK-4 responded to bacterial infection in the intestine by an enhancement of its expression level. These results indicate that PmIRAK-4 may play a role at least in the intestinal innate immunity of P. monodon.
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| 6. |
- Watthanasurorot, Apiruck, et al.
(författare)
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An ancient cytokine, astakine, mediates circadian regulation of invertebrate hematopoiesis
- 2011
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Ingår i: Cellular and Molecular Life Sciences (CMLS). - : Springer Science and Business Media LLC. - 1420-682X .- 1420-9071. ; 68:2, s. 315-323
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Tidskriftsartikel (refereegranskat)abstract
- Invertebrate circulating hemocytes are key players in the innate immune defense and their continuous renewal from hematopoietic tissues is tightly regulated in crustaceans by astakine, a new family of cytokines sharing a prokineticin (PROK) domain. In vertebrates, brain PROKs function as transmitters of circadian rhythms and we present evidence that hemocyte release from hematopoietic tissues in crayfish is under circadian regulation, a direct result of rhythmic expression of astakine. We demonstrate that the observed variation in astakine expression has an impact on innate immunity assessed as susceptibility to a pathogenic Pseudomonas species. These findings enlighten the importance of comparing immune responses at fixed times not to neglect circadian regulation of innate immunity. Moreover, our results entail an evolutionary conserved function for prokineticins as mediators of circadian rhythm, and for the first time show a role for this domain in circadian regulation of hematopoiesis that may have implications also in vertebrates.
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| 7. |
- Watthanasurorot, Apiruck, et al.
(författare)
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Astakine 2-the Dark Knight Linking Melatonin to Circadian Regulation in Crustaceans
- 2013
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Ingår i: PLOS Genetics. - : Public Library of Science (PLoS). - 1553-7390 .- 1553-7404. ; 9:3, s. e1003361-
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Tidskriftsartikel (refereegranskat)abstract
- Daily, circadian rhythms influence essentially all living organisms and affect many physiological processes from sleep and nutrition to immunity. This ability to respond to environmental daily rhythms has been conserved along evolution, and it is found among species from bacteria to mammals. The hematopoietic process of the crayfish Pacifastacus leniusculus is under circadian control and is tightly regulated by astakines, a new family of cytokines sharing a prokineticin (PROK) domain. The expression of AST1 and AST2 are light-dependent, and this suggests an evolutionarily conserved function for PROK domain proteins in mediating circadian rhythms. Vertebrate PROKs are transmitters of circadian rhythms of the suprachiasmatic nucleus (SCN) in the brain of mammals, but the mechanism by which they function is unknown. Here we demonstrate that high AST2 expression is induced by melatonin in the brain. We identify RACK1 as a binding protein of AST2 and further provide evidence that a complex between AST2 and RACK1 functions as a negative-feedback regulator of the circadian clock. By DNA mobility shift assay, we showed that the AST2-RACK1 complex will interfere with the binding between BMAL1 and CLK and inhibit the E-box binding activity of the complex BMAL1-CLK. Finally, we demonstrate by gene knockdown that AST2 is necessary for melatonin-induced inhibition of the complex formation between BMAL1 and CLK during the dark period. In summary, we provide evidence that melatonin regulates AST2 expression and thereby affects the core clock of the crustacean brain. This process may be very important in all animals that have AST2 molecules, i.e. spiders, ticks, crustaceans, scorpions, several insect groups such as Hymenoptera, Hemiptera, and Blattodea, but not Diptera and Coleoptera. Our findings further reveal an ancient evolutionary role for the prokineticin superfamily protein that links melatonin to direct regulation of the core clock gene feedback loops.
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| 8. |
- Watthanasurorot, Apiruck, et al.
(författare)
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Bacteria-Induced Dscam Isoforms of the Crustacean, Pacifastacus leniusculus
- 2011
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Ingår i: PLoS Pathogens. - : Public Library of Science (PLoS). - 1553-7366 .- 1553-7374. ; 7:6
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Tidskriftsartikel (refereegranskat)abstract
- The Down syndrome cell adhesion molecule, also known as Dscam, is a member of the immunoglobulin super family. Dscam plays an essential function in neuronal wiring and appears to be involved in innate immune reactions in insects. The deduced amino acid sequence of Dscam in the crustacean Pacifastacus leniusculus (PlDscam), encodes 9(Ig)-4(FNIII)-(Ig)-2(FNIII)-TM and it has variable regions in the N-terminal half of Ig2 and Ig3 and the complete Ig7 and in the transmembrane domain. The cytoplasmic tail can generate multiple isoforms. PlDscam can generate more than 22,000 different unique isoforms. Bacteria and LPS injection enhanced the expression of PlDscam, but no response in expression occurred after a white spot syndrome virus (WSSV) infection or injection with peptidoglycans. Furthermore, PlDscam silencing did not have any effect on the replication of the WSSV. Bacterial specific isoforms of PlDscam were shown to have a specific binding property to each tested bacteria, E. coli or S. aureus. The bacteria specific isoforms of PlDscam were shown to be associated with bacterial clearance and phagocytosis in crayfish.
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| 9. |
- Watthanasurorot, Apiruck, et al.
(författare)
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Hijacking of Host Calreticulin Is Required for the White Spot Syndrome Virus Replication Cycle
- 2014
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Ingår i: Journal of Virology. - 0022-538X .- 1098-5514. ; 88:14, s. 8116-8128
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Tidskriftsartikel (refereegranskat)abstract
- We have previously shown that multifunctional calreticulin (CRT), which resides in the endoplasmic reticulum (ER) and is involved in ER-associated protein processing, responds to infection with white spot syndrome virus (WSSV) by increasing mRNA and protein expression and by forming a complex with gC1qR and thereby delaying apoptosis. Here, we show that CRT can directly interact with WSSV structural proteins, including VP15 and VP28, during an early stage of virus infection. The binding of VP28 with CRT does not promote WSSV entry, and CRT-VP15 interaction was detected in the viral genome in virally infected host cells and thus may have an effect on WSSV replication. Moreover, CRT was detected in the viral envelope of purified WSSV virions. CRT was also found to be of high importance for proper oligomerization of the viral structural proteins VP26 and VP28, and when CRT glycosylation was blocked with tunicamycin, a significant decrease in both viral replication and assembly was detected. Together, these findings suggest that CRT confers several advantages to WSSV, from the initial steps of WSSV infection to the assembly of virions. Therefore, CRT is required as a “vital factor” and is hijacked by WSSV for its replication cycle.IMPORTANCE White spot syndrome virus (WSSV) is a double-stranded DNA virus and the cause of a serious disease in a wide range of crustaceans that often leads to high mortality rates. We have previously shown that the protein calreticulin (CRT), which resides in the endoplasmic reticulum (ER) of the cell, is important in the host response to the virus. In this report, we show that the virus uses this host protein to enter the cell and to make the host produce new viral structural proteins. Through its interaction with two viral proteins, the virus “hijacks” host calreticulin and uses it for its own needs. These findings provide new insight into the interaction between a large DNA virus and the host protein CRT and may help in understanding the viral infection process in general.
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