Zebrafish in Inflammasome Research

Forn-Cuní, Gabriel; Meijer, Annemarie H.; Varela, Mónica

doi:10.3390/cells8080901

Cited by 36 publications

(28 citation statements)

References 91 publications

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“…Pyroptotic cell death is associated with the activity of inflammatory caspases, like Caspase 1 and Caspase 4/5/11, and is characterized by the formation of Gasdermin pores in the cell membrane [36][37][38] . We have recently found that Caspase a (Caspa) is the Caspase-family member that induces pyroptosis of Mm-infected macrophages in zebrafish via Gasdermin Eb (Gsdmeb) 39,40 . Thus, we analyzed Caspa activity at 2dpf, the time point where we observed increased cell death in dram1 Δ19n/Δ19n .…”

Section: Dram1 Deficiency Results In Increased Pyroptotic Cell Death mentioning

confidence: 99%

Deficiency in the autophagy modulator Dram1 exacerbates pyroptotic cell death of Mycobacteria-infected macrophages

et al. 2020

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DNA damage regulated autophagy modulator 1 (DRAM1) is a stress-inducible regulator of autophagy and cell death. DRAM1 has been implicated in cancer, myocardial infarction, and infectious diseases, but the molecular and cellular functions of this transmembrane protein remain poorly understood. Previously, we have proposed DRAM1 as a host resistance factor for tuberculosis (TB) and a potential target for host-directed anti-infective therapies. In this study, we generated a zebrafish dram1 mutant and investigated its loss-of-function effects during Mycobacterium marinum (Mm) infection, a widely used model in TB research. In agreement with previous knockdown analysis, dram1 mutation increased the susceptibility of zebrafish larvae to Mm infection. RNA sequencing revealed major effects of Dram1 deficiency on metabolic, immune response, and cell death pathways during Mm infection, and only minor effects on proteinase and metabolic pathways were found under uninfected conditions. Furthermore, unchallenged dram1 mutants did not display overt autophagic defects, but autophagic targeting of Mm was reduced in the absence of Dram1. The phagocytic ability of macrophages in dram1 mutants was unaffected, but acidification of Mm-containing vesicles was strongly reduced, indicating that Dram1 is required for phagosome maturation. By in vivo imaging, we observed that Dram1-deficient macrophages fail to restrict Mm during early stages of infection. The resulting increase in bacterial burden could be reverted by knockdown of inflammatory caspase a (caspa) and gasdermin Eb (gsdmeb), demonstrating pyroptosis as the mechanism underlying premature cell death of Mm-infected macrophages in dram1 mutants. Collectively, these data demonstrate that dissemination of mycobacterial infection in zebrafish larvae is promoted in the absence of Dram1 due to reduced maturation of mycobacteria-containing vesicles, failed intracellular containment, and consequent pyroptotic death of infected macrophages. These results provide new evidence that Dram1 plays a central role in host resistance to intracellular infection, acting at the crossroad of autophagy and cell death.

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Section: Dram1 Deficiency Results In Increased Pyroptotic Cell Death mentioning

confidence: 99%

Deficiency in the autophagy modulator Dram1 exacerbates pyroptotic cell death of Mycobacteria-infected macrophages

et al. 2020

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“…The large p20 and small p10 domains of the 35 putative C. gigas caspases identified by a BLAST search, together with the 23 previously identified caspases in bivalves, were aligned with caspases homologues (caspase-1 -caspase-10) of vertebrates and other invertebrate species using the default parameter of MUSCLE v3.8.31 [85] and edited manually in case of errors. Representatives of caspase homologues to vertebrate caspase-11caspase-16 or tunicates caspase-17caspase-22 were excluded as previous research has shown that these caspases are specific to these subphyla [86,87]. For those putative caspases, which did not contain a small p10 domain, remaining protein sequence downstream to the large p20 domain were used in the alignment instead.…”

Section: Identification Bivalve Caspases and Phylogenetic Analysismentioning

confidence: 99%

Phylogenetic analysis of the caspase family in bivalves: implications for programmed cell death, immune response and development

Vogeler

Carboni

et al. 2021

BMC Genomics

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Background Apoptosis is an important process for an organism’s innate immune system to respond to pathogens, while also allowing for cell differentiation and other essential life functions. Caspases are one of the key protease enzymes involved in the apoptotic process, however there is currently a very limited understanding of bivalve caspase diversity and function. Results In this work, we investigated the presence of caspase homologues using a combination of bioinformatics and phylogenetic analyses. We blasted the Crassostrea gigas genome for caspase homologues and identified 35 potential homologues in the addition to the already cloned 23 bivalve caspases. As such, we present information about the phylogenetic relationship of all identified bivalve caspases in relation to their homology to well-established vertebrate and invertebrate caspases. Our results reveal unexpected novelty and complexity in the bivalve caspase family. Notably, we were unable to identify direct homologues to the initiator caspase-9, a key-caspase in the vertebrate apoptotic pathway, inflammatory caspases (caspase-1, − 4 or − 5) or executioner caspases-3, − 6, − 7. We also explored the fact that bivalves appear to possess several unique homologues to the initiator caspase groups − 2 and − 8. Large expansions of caspase-3 like homologues (caspase-3A-C), caspase-3/7 group and caspase-3/7-like homologues were also identified, suggesting unusual roles of caspases with direct implications for our understanding of immune response in relation to common bivalve diseases. Furthermore, we assessed the gene expression of two initiator (Cg2A, Cg8B) and four executioner caspases (Cg3A, Cg3B, Cg3C, Cg3/7) in C. gigas late-larval development and during metamorphosis, indicating that caspase expression varies across the different developmental stages. Conclusion Our analysis provides the first overview of caspases across different bivalve species with essential new insights into caspase diversity, knowledge that can be used for further investigations into immune response to pathogens or regulation of developmental processes.

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“…The large p20 and small p10 domains of the 35 putative C. gigas caspases identi ed by a BLAST search, together with the 23 previously identi ed caspases in bivalves, were aligned with caspases homologs (caspase-1 -caspase-10) of vertebrates and other invertebrate species using the default parameter of MUSCLE v3.8.31 (101) and edited manually in case of errors. Representatives of caspase homologs to vertebrate caspase-11 -caspase-16 or tunicates caspase-17 -caspase-22 were excluded as previous research has shown that these caspases are speci c to these subphyla (102,103). For those putative caspases, which did not contain a small p10 domain, remaining protein sequence downstream to the large p20 domain were used in the alignment instead.…”

Section: Methodsmentioning

confidence: 99%

Phylogenetic analysis of the caspase family in bivalves: implications for programmed cell death, immune response and development

Vogeler

Carboni

et al. 2020

Preprint

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Background: Apoptosis is an important process for an organism’s innate immune system to respond to pathogens, while also allowing for cell differentiation and other essential life functions. Caspases are one of the key protease enzymes involved in the apoptotic process, however there is currently a very limited understanding of bivalve caspase diversity and functions. Results: In this work, we investigated the presence of caspase homologs using a combination of bioinformatics and phylogenetic analyses. We blasted the Crassostrea gigas genome for caspase homologs and identified thirty-five potential homologs in the addition to the already cloned twenty-three bivalve caspases. As such, we present information about the phylogenetic relationship of all identified bivalve caspases in relation to their homology to well-established vertebrate and invertebrate caspases. Our results reveal unexpected novelty and complexity in the bivalve caspase family. Notably, we were unable to identify direct homologues to the initiator caspase-9, a key-caspase in the vertebrate apoptotic pathway, inflammatory caspases (caspase-1,-4 or -5) or executioner caspases-3, -6, -7. We also explored the fact that bivalves appear to possess several unique homologs to the initiator caspase groups -2 and -8. Large expansions of caspase-3 like homologues (caspase-3A-C), caspase-3/7 group and caspase-3/7-like homologues were also identified, suggesting unusual roles of caspases with direct implications for our understanding of immune response in relation to common bivalve diseases. Furthermore, we assessed the gene expression of two initiator (Cg2A, Cg8B) and four executioner caspases (Cg3A, Cg3B, Cg3C, Cg3/7) in C. gigas late-larval development and during metamorphosis, indicating that caspase expressions vary across the different developmental stages.Conclusion: Our analysis provides the first overview of caspases across different bivalve species with essential new insight into caspase diversity, knowledge that can be used for further investigations into response to pathogens or regulation of developmental processes.

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Zebrafish in Inflammasome Research

Cited by 36 publications

References 91 publications

Deficiency in the autophagy modulator Dram1 exacerbates pyroptotic cell death of Mycobacteria-infected macrophages

Deficiency in the autophagy modulator Dram1 exacerbates pyroptotic cell death of Mycobacteria-infected macrophages

Phylogenetic analysis of the caspase family in bivalves: implications for programmed cell death, immune response and development

Phylogenetic analysis of the caspase family in bivalves: implications for programmed cell death, immune response and development

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