In silico analysis and effects of environmental salinity in the expression and activity of digestive α-amylase and trypsins from the euryhaline crab Neohelice granulata

MARTÍNEZ-RODRÍGUEZ, GONZALO; LÓPEZ MAÑANES, ALEJANDRA; MARTOS-SITCHA, JUAN ANTONIO; MANCERA, JUAN MIGUEL; ASARO, ANTONELA; MARTÍNEZ-RODRÍGUEZ, GONZALO; LÓPEZ MAÑANES, ALEJANDRA; MARTOS-SITCHA, JUAN ANTONIO; MANCERA, JUAN MIGUEL; ASARO, ANTONELA

CANADIAN JOURNAL OF ZOOLOGY

NATL RESEARCH COUNCIL CANADA-N R C RESEARCH PRESS

Año: 2018 vol. 96 p. 127 - 139

0008-4301

Studies on molecular characteristics and modulation of expression of α-amylase and trypsin in the41 hepatopancreas of intertidal euryhaline crabs are lacking. In this work, we cloned and studied by in silico42 approaches the characteristics of cDNA sequences for α-amylase and two trypsins isoforms, as well as the43 effect of environmental salinity on gene expression and protein activities in hepatopancreas of Neohelice44 granulata (Dana,1852), as a good invertebrate model species. The cDNA sequence of α-amylase is 1,63745 bp long, encoding 459 amino acid residues. Trypsin 1 and 2 are 689 bp and 1,174 bp long, encoding 20446 and 151 amino acid residues, respectively. Multiple sequence alignment of deduced protein sequences47 revealed the presence of conserved motifs found in other invertebrates. In crabs acclimated at 37 psu48 (hypo-regulation), α-amylase mRNA level and total pancreatic amylase activity were higher than at 1049 psu (hyper-regulation) and 35 psu (osmoconformation). Trypsin 1 mRNA levels increased at 37 psu while50 trypsin 2 levels decreased at 10 and 37 psu. Total trypsin activity was similar in all salinities. Our results51 showed a differential modulation of α-amylase and trypsins expression and total amylase activity by52 salinity acclimation, suggesting the occurrence of distinct mechanisms of regulation at different levels53 that could lead to digestive adjustments in relation to hyper- and/or hypo-regulation.