1. In silico analysis and effects of environmental salinity in the expression and activity of digestive [alpha]-amylase and trypsins from the euryhaline crab Neohelice granulata
- Author
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Asaro, Antonela, Martos-Sitcha, Juan Antonio, Martinez-Rodriguez, Gonzalo, Mancera, Juan Miguel, and Mananes, Alejandra Antonia Lopez
- Subjects
Trypsin -- Measurement ,Amylases -- Measurement ,Salinity -- Physiological aspects ,Varunidae -- Physiological aspects -- Environmental aspects ,Zoology and wildlife conservation - Abstract
Studies on molecular characteristics and modulation of expression of [alpha]-amylase and trypsin in the hepatopancreas of intertidal euryhaline crabs are lacking. In this work, we cloned and studied by in silico approaches the characteristics of cDNA sequences for [alpha]-amylase and two trypsins isoforms, as well as the effect of environmental salinity, on gene expression and protein activities in the hepatopancreas of Neohelice granulata (Dana, 1851), which is a good invertebrate model species. The cDNA sequence of [alpha]-amylase is 1637 bp long, encoding 459 amino acid residues. Trypsin 1 and 2 are 689 and 1174 bp long, encoding 204 and 151 amino acid residues, respectively. Multiple sequence alignment of deduced protein sequences revealed the presence of conserved motifs found in other invertebrates. In crabs acclimated at 37 psu (hyporegulation), [alpha]-amylase mRNA level and total pancreatic amylase activity were higher than at 10 psu (hyperregulation) and 35 psu (osmoconformation). Trypsin 1 mRNA levels increased at 37 psu, while trypsin 2 levels decreased at 10 and 37 psu. Total trypsin activity was similar in all salinities. Our results showed a differential modulation of [alpha]-amylase and trypsin expression and total amylase activity by salinity acclimation, suggesting the occurrence of distinct mechanisms of regulation at different levels that could lead to digestive adjustments in relation to hyperregulation and (or) hyporegulation. Key words: [alpha]-amylase, trypsin, cloning, digestive flexibility, hepatopancreas, Neohelice granulata. Les etudes des caracteristiques moleculaires et de la modulation de l'expression de l'[alpha]-amylase et de la trypsine dans l'hepatopancreas de crabes euryhalins intertidaux sont rares. Nous avons clone et etudie par des approches in silico les caracteristiques de sequences d'ADNc pour l'[alpha]-amylase et deux isoformes de la trypsine, ainsi que l'effet de la salinite ambiante sur l'expression genique et l'activite proteique dans l'hepatopancreas de Neohelice granulata (Dana, 1851), une bonne espece modele d'invertebre. La longueur de la sequence d'ADNc de l'[alpha]-amylase fait 1637 pb, codant 459 residus d'aminoacides. Les trypsines 1 et 2 font 689 et 1174 bp de longueur, codant 204 et 151 residus d'aminoacides, respectivement. L'alignement de sequences multiples de sequences proteiques deduites revele la presence de motifs conserves presents chez d'autres invertebres. Chez des crabes acclimates a 37 psu (hyporegulation), la concentration d'ARNm de l'[alpha]-amylase et l'activite de l'amylase pancreatique totale etaient plus elevees qu'a 10 psu (hyperregulation) et qu'a 35 psu (osmoconformation). Les concentrations d'ARNm de la trypsine 1 augmentaient a 37 psu, alors que les concentrations de trypsine 2 diminuaient a 10 et a 37 psu. L'activite totale des trypsines etait semblable a toutes les salinites. Nos resultats font ressortir une modulation differentielle de l'expression de l'[alpha]-amylase et des trypsines et de l'activite de l'amylase totale par l'acclimatation a la salinite, ce qui indiquerait l'intervention de differents mecanismes de regulation selon la concentration pouvant se traduire par des ajustements digestifs associes a l'hyperregulation et l'hyporegulation. [Traduit par la Redaction] Mots-cles: [alpha]-amylase, trypsine, clonage, souplesse digestive, hepatopancreas, Neohelice granulata., Introduction Phenotypic flexibility involves changes in attributes atdifferent levels (from molecular to organismal) in response to variations in external stimuli. These reversible changes can increase probability of survival particularly in [...]
- Published
- 2018
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