Abstract
Crustaceans are major constituents of aquatic ecosystems and, as such, changes in their behavior and the structure and function of their bodies can serve as indicators of alterations in their immediate environment, such as those associated with climate change and anthropogenic contamination. We have used bioinformatics and a de novo transcriptome assembly approach to identify potential targets for developing specific antibodies to serve as nervous system function markers for freshwater prawns of the Macrobrachium spp. Total RNA was extracted from brain ganglia of Macrobrachium carcinus freshwater prawns and Illumina Next Generation Sequencing was performed using an Eel Pond mRNA Seq Protocol to construct a de novo transcriptome. Sequencing yielded 97,202,662 sequences: 47,630,546 paired and 1,941,570 singletons. Assembly with Trinity resulted in 197,898 assembled contigs from which 30,576 were annotated: 9,600 by orthology, 17,197 by homology, and 3,779 by transcript families. We looked for glutamate receptors contigs, due to their main role in crustacean excitatory neurotransmission, and found 138 contigs related to ionotropic receptors, 32 related to metabotropic receptors, and 18 to unidentified receptors. After performing multiple sequence alignments within different biological organisms and antigenicity analysis, we were able to develop antibodies for prawn AMPA ionotropic glutamate receptor 1, metabotropic glutamate receptor 1 and 4, and ionotropic NMDA glutamate receptor subunit 2B, with the expectation that the availability of these antibodies will help broaden knowledge regarding the underlying structural and functional mechanisms involved in prawn behavioral responses to environmental impacts. The Macrobrachium carcinus brain transcriptome can be an important tool for examining changes in many other nervous system molecules as a function of developmental stages, or in response to particular conditions or treatments.
Highlights
Crustaceans are major constituents of aquatic ecosystems, living in different habitats depending on their needs, and playing an ecological role in their habitat as shredders and nutrient cyclers [1]
We looked for glutamate receptors contigs, due to their main role in crustacean excitatory neurotransmission, and found 138 contigs related to ionotropic receptors, 32 related to metabotropic receptors, and 18 to unidentified receptors
Four adult male Macrobrachium carcinus (M. carcinus) freshwater prawns were caught in non-urban rivers in Yabucoa, Puerto Rico, as authorized through permit 2016-IC-145 (R-VS-PVS15-SJ-00560–08092016) of the Department of Environmental and Natural Resources of the Government of Puerto Rico
Summary
Crustaceans are major constituents of aquatic ecosystems, living in different habitats depending on their needs, and playing an ecological role in their habitat as shredders and nutrient cyclers [1]. They are beneficial to the ecosystem as they participate in consuming decaying vegetables and animal bodies in the water [2]. Indicators of environmental stress in water include relative abundance and diversity, feeding activity, drifting, changes in metabolism, molting and growth, immune functions, reproductive capacity and locomotion [4]. Characterization of mechanisms and molecules associated with behavioral and metabolic or physiological changes is a current research focus given the commercial and environmental relevance of these invertebrate animals
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