Abstract
BackgroundThe giant freshwater prawn, Macrobrachium rosenbergii, is a decapod crustacean that is commercially important as a food source. Farming of commercial crustaceans requires an efficient management strategy because the animals are easily subjected to stress and diseases during the culture. Autophagy, a stress response process, is well-documented and conserved in most animals, yet it is poorly studied in crustaceans.ResultsIn this study, we have performed an in silico search for transcripts encoding autophagy-related (Atg) proteins within various tissue transcriptomes of M. rosenbergii. Basic Local Alignment Search Tool (BLAST) search using previously known Atg proteins as queries revealed 41 transcripts encoding homologous M. rosenbergii Atg proteins. Among these Atg proteins, we selected commonly used autophagy markers, including Beclin 1, vacuolar protein sorting (Vps) 34, microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B), p62/sequestosome 1 (SQSTM1), and lysosomal-associated membrane protein 1 (Lamp-1) for further sequence analyses using comparative alignment and protein structural prediction. We found that crustacean autophagy marker proteins contain conserved motifs typical of other animal Atg proteins. Western blotting using commercial antibodies raised against human Atg marker proteins indicated their presence in various M. rosenbergii tissues, while immunohistochemistry localized Atg marker proteins within ovarian tissue, specifically late stage oocytes.ConclusionsThis study demonstrates that the molecular components of autophagic process are conserved in crustaceans, which is comparable to autophagic process in mammals. Furthermore, it provides a foundation for further studies of autophagy in crustaceans that may lead to more understanding of the reproduction- and stress-related autophagy, which will enable the efficient aquaculture practices.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2996-4) contains supplementary material, which is available to authorized users.
Highlights
The giant freshwater prawn, Macrobrachium rosenbergii, is a decapod crustacean that is commercially important as a food source
Gene mining of autophagy-related proteins in M. rosenbergii transcriptomes An in silico search of Atg proteins in M. rosenbergii transcriptomes revealed 41 transcripts predicted to encode Atg proteins (Table 1)
The first category includes the proteins involved in formation of the autophagy interactome that initiates the process of autophagy; the second category consists of proteins that are responsible for Atg9-WIPI1 complex formation, which triggers membrane nucleation and phagophore formation; the third category includes proteins that are involved in Atg12 conjugation, which mediates phagophore elongation; the fourth category comprises of proteins involved in Atg8/LC3 conjugation, which controls cargo sequestration and autophagosome formation; and the last category includes the phosphatidylinositol 3-phosphate (PI3P)-related proteins with other proteins associated with autophagic activities
Summary
The giant freshwater prawn, Macrobrachium rosenbergii, is a decapod crustacean that is commercially important as a food source. A stress response process, is well-documented and conserved in most animals, yet it is poorly studied in crustaceans. Macroautophagy (autophagy) is a complex process involving degradation of cellular constituents to maintain organelle and protein homeostasis in response to various stresses, including genomic, oxidative, hypoxic, proteotoxic and metabolic stresses as well as starvation [1]. Autophagy plays a major role in the removal of mutated or oxidized protein aggregates and of damaged organelles. It is involved in restoring nutritional and metabolic imbalances during starvation. Autophagy is an evolutionary conserved mechanism ubiquitously present in all eukaryotic cells. It is driven and regulated by several proteins designated as autophagyrelated (Atg) proteins (ATG in mammals). Many Atg orthologs have subsequently been identified in higher eukaryotes, including
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