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Title: Transcriptomic analysis of the autophagy machinery in crustaceans
Authors: Saowaros Suwansa-ard
Wilairat Kankuan
Tipsuda Thongbuakaew
Jirawat Saetan
Napamanee Kornthong
Thanapong Kruangkum
Kanjana Khornchatri
Cummins, Scott F.
Isidoro, Ciro
Prasert Sobhon
Mahidol University. Faculty of Science. Department of Anatomy
Keywords: Open Access article;Autophagy;Crustaceans;Transcriptome;Nervous system;Ovary;Autophagy markers
Issue Date: 2016
Citation: BMC Genomics. Vol. 17, (2016), 587
Abstract: Background: The 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. Results: In 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. Conclusions: This 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.
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