Molecular cloning and localization of GABAA receptor-associated protein in the rotifer Brachionus plicatilis
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γ-Aminobutyric acid receptor type A-associated protein (GABARAP) and its homologs constitute a protein family found in many eukaryotes from yeast to human, and are known to be involved in intracellular membrane trafficking of GABAA receptors and autophagy. In this study, we cloned cDNA-encoding GABARAP from the monogonont rotifer Brachionus plicatilis and examined for its tissue distribution at the protein level in neonates, males and females. Using reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE) techniques, we showed that like other GABARAPs, rotifer GABARAP was also composed of 117 amino acids and highly homologous to vertebrate GABARAP2 ortholog (74–76% identity). GABARAP was demonstrated with its specific antibody to be ubiquitously distributed, irrespective of neonates, males, and females, in the coronal area that covers brain and contains most mechano- and chemoreceptors. Rotifer GABARAP was also expressed in the mature eggs but not in immature eggs. Double immunostaining with mammalian anti-GABA γ receptor antibody showed that rotifer GABARAP co-localized with GABA receptor, suggesting the association of the two proteins. The presence of GABARAP in rotifer implies that it is highly conserved during evolution, and plays important roles in various biological processes.
CitationMarcial, H. S., Suga, K., Kinoshita, S., Kaneko, G., Hagiwara, A., & Watabe, S. (2014). Molecular cloning and localization of GABAA receptor-associated protein in the rotifer Brachionus plicatilis.
Chemoreceptors; Antibodies; Marine invertebrates; Brain; Receptors; Proteins; Transcription; Hydrobiology; Evolution; Amino acids; Eggs; Reverse transcription; gamma -Aminobutyric acid receptors; GABARAP protein; Reviews; gamma -Aminobutyric acid A receptors; Membrane trafficking; Neonates; Phagocytosis; Yeasts; Amino acids; Membranes; Acids; Rotifera; Brachionus plicatilis; Rotifera
This study was partly supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS).
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Book chapterOM Millamena - In OM Millamena, RM Coloso & FP Pascual (Eds.), Nutrition in Tropical Aquaculture: Essentials of fish nutrition, feeds, and feeding of tropical aquatic species, 2002 - Aquaculture Department, Southeast Asian Fisheries Development CenterThis section aims to teach the reader the ten essential amino acids required by fish and their chemical structures, distinguish between essential and non-essential amino acids; the fate of absorbed amino acids in fish; effects of deficiencies and excesses of dietary amino acids in fish diets; the procedure on how to determine the qualitative and quantitative amino acid requirements of fish; methods of evaluating protein quality; and how to determine protein requirements of some aquaculture species.
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