Autophagic processes in Mytilus galloprovincialis hemocytes: Effects of Vibrio tapetis

Balbi, Teresa; Cortese, Katia; Ciacci, Caterina; Bellese, Grazia; Vezzulli, Luigi; Pruzzo, Carla; Canesi, Laura

doi:10.1016/j.fsi.2017.12.003

Cited by 31 publications

(37 citation statements)

References 46 publications

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“…The results showed that autophagy is constitutively detected at both the molecular and protein levels in the tested tissues of the Pacific oyster. Several studies have already reported that autophagy is implicated in a variety of cellular processes like innate immunity, response to environmental stressors (like anoxia and hyperthermia), and embryonic and larval development in mollusks [11,12,15,16,44]. However, in C. gigas, the cellular functions of autophagy need to be further investigated.…”

Section: Discussionmentioning

confidence: 99%

Identification of the autophagy pathway in a mollusk bivalve, Crassostrea gigas

et al. 2020

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The Pacific oyster, Crassostrea gigas, is a mollusk bivalve commercially important as a food source. Pacific oysters are subjected to stress and diseases during culture. The autophagy pathway is involved in numerous cellular processes, including responses to starvation, cell death, and microorganism elimination. Autophagy also exists in C. gigas, and plays a role in the immune response against infections. Although this process is well-documented and conserved in most animals, it is still poorly understood in mollusks. To date, no study has provided a complete overview of the molecular mechanism of autophagy in mollusk bivalves. In this study, human and yeast ATG protein sequences and public databases (Uniprot and NCBI) were used to identify protein members of the C. gigas autophagy pathway. A total of 35 autophagy related proteins were found in the Pacific oyster. RACE-PCR was performed on several genes. Using molecular (real-time PCR) and protein-based (western blot and immunohistochemistry) approaches, the expression and localization of ATG12, ATG9, BECN1, MAP1LC3, MTOR, and SQSTM1, was investigated in different tissues of the Pacific oyster. Comparison with human and yeast counterparts demonstrated a high homology with the human autophagy pathway. The results also demonstrated that the key autophagy genes and their protein products were expressed in all the analyzed tissues of C. gigas. This study allows the characterization of the complete C. gigas autophagy pathway for the first time.

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Section: Discussionmentioning

confidence: 99%

Identification of the autophagy pathway in a mollusk bivalve, Crassostrea gigas

et al. 2020

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“…Hemocyte monolayers were prepared as previously described [18,20] and incubated at 18°C with suspensions of V. coralliilyticus suitably diluted in hemolymph serum at different concentrations (5 x 5 , 5 x 10 6 , 5 x 10 7 CFU/mL), for different periods of times, depending on the endpoint measured. Untreated hemocyte samples in serum were run in parallel.…”

Section: In Vitro Challenge Of Mytilus Hemocytes With V Coralliilyticusmentioning

confidence: 99%

“…Lysosomal membrane stability-LMS in hemocyte monolayers was evaluated by the NRRT assay as previously described [17,18,20]. Hemocyte monolayers on glass slides were pre-incubated for 30 min with different concentrations of V. coralliilyticus.…”

Section: Determination Of Lysosomal Membrane Stability-lmsmentioning

confidence: 99%

“…Although Mytilus spp., including M. galloprovincialis, is particularly resistant to bacterial infections, it shows a remarkable specificity of the immune response towards different Vibrio spp. and strains, as demonstrated both in vitro and in vivo studies in adults [2,17,18]. In contrast, little information is available on the possible vibrio pathogens affecting Mytilus embryo development [19].…”

Section: Introductionmentioning

confidence: 99%

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Responses of Mytilus galloprovincialis to challenge with the emerging marine pathogen Vibrio coralliilyticus

Balbi

Auguste

Cortese

et al. 2019

Fish & Shellfish Immunology

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Vibrio coralliilyticus (V.c.) has emerged as a coral pathogen of concern throughout the Indo-Pacific reef. The interest towards understanding its ecology and pathogenic potential has increased since V.c. was shown to be strongly virulent also for other species; in particular, it represents a serious threat for bivalve aquaculture, being one of the most important emerging pathogen responsible for oyster larval mortalities worldwide. V.c. has a tightly regulated temperature-dependent virulence and it has been related to mass mortalities events of benthic invertebrates also in the temperate northwestern Mediterranean Sea. However, no data are available on the effects of V.c. in the mussel Mytilus galloprovincialis, the most abundant aquacultured species in this area. In this work, responses of M. galloprovincialis to challenge with V.c. (ATCC BAA-450) were investigated. In vitro, short term responses of mussel hemocytes were evaluated in terms of lysosomal membrane stability, bactericidal activity, lysozyme release, ROS and NO production, and ultrastructural changes, evaluated by TEM. In vivo, hemolymph parameters were measured in mussels challenged with V.c. at 24h p.i. Moreover, the effects of V.c. on mussel early embryo development (at 48 hpf) were evaluated. The results show that both in vitro and in vivo, mussels were unable to activate immune response towards V.c., and that challenge mainly induced lysosomal stress in the hemocytes. Moreover, V.c. showed a strong and concentration-dependent embryotoxicity. Overall, the results indicate that, although M. galloprovincialis is considered a resistant species to vibrio infections, the emerging pathogen V.c. can represent a potential threat to mussel aquaculture.

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“…In mussels infected by V. anguillarum, disruption in energy metabolism and induction of oxidative stress were revealed (Ji et al, 2013), whereas immune stress, osmotic disruption and reduced energy demand were observed in V. harveyi-challenged M. galloprovincialis (Liu et al, 2014). Interestingly, autophagic processes were induced by V. tapetis in hemocytes of M. galloprovincialis, as a result of protective mechanisms towards damaged cell components (Balbi et al, 2018). More recently, a dose-dependent decrease in hemocyte lysosomal membrane stability and bactericidal activity of whole hemolymph, were observed in M. galloprovincialis in response to in vitro and in vivo challenge with the emerging marine pathogen V. coralliilyticus (Balbi et al, 2019).…”

Section: Introductionmentioning

confidence: 99%