This study constitutes a first attempt to investigate intraspecific differences in osmoregulatory capacity and digestive and metabolic responses at the biochemical level in relation to hyper-and hypo-regulation in a single species of estuarine crab inhabiting contrasting habitats within a same intertidal area. We compared hemolymph osmolality, key digestive enzymes, glycemia and energy reserves in Neohelice granulata (Dana in Proc Acad Nat Sci Philadelphia 5:247-254, 1851) from the mudflat and saltmarsh of Mar Chiquita coastal lagoon (37°32 0 /37°45 0 S-57°19 0 /57°26 0 W) under a wide range of salinities (6-60 psu). Individuals from both sites exhibited high and similar osmoregulatory capacity, but while in individuals from mudflat low and high salinities affected lipase activity in hepatopancreas and triglycerides in muscle, in crabs from saltmarsh, high salinities affected glycogen in anterior gills. Low salinity differentially affected free glucose in anterior gills. The results suggest the occurrence of intraspecific distinct digestive and metabolic adjustments in relation to osmoregulatory responses and habitat.

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Strikingly, in spite of its physiological importance, information about occurrence, biochemical characteristics and mechanisms of regulation of aminopeptidase-N (APN) in the hepatopancreas of intertidal euryhaline crabs is still lacking. In this work, we determined the occurrence, biochemical characteristics, response to environmental salinity and dopamine of APN in the hepatopancreas of the euryhaline crab Neohelice granulata (Dana 1851) from the open mudflat of Mar Chiquita coastal lagoon (Buenos Aires province, Argentina). APN activity was maximal at pH and temperature range of 7.6-9.0 and 37-45 °C, respectively. APN activity exhibited Michaelis-Menten kinetics (apparent Km = 0.19 ± 0.10 mM) (pH 7.6, 37 °C) and appeared to be sensitive to bestatin (I 50 = 15 mM) and EDTA (I 50 = 9 mM). In crabs acclimated to 10 psu (hyper-regulation conditions) and 37 psu (hypo-regulation conditions), APN activity was about 45 and 160% higher, respectively, than in 35 psu (osmoconformation). APN activity in the hepatopancreas was stimulated in vitro (about 137%) by 10(-4) M dopamine. Higher dopamine concentrations produced a similar extent of increase. The responses of APN activity to salinity and dopamine in vitro suggest the role of APN in digestive adjustments upon hyper and hypo-regulatory conditions and its modulation via direct mechanisms on hepatopancreas by dopamine.