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Osmotic effects on arginine kinase function in living muscle of the blue crab Callinectes sapidus

Department of Biological Sciences, University of North Carolina at
Wilmington, 601 South College Road, Wilmington, NC 28403-3297, USA
* Present address: Department of Biology, University of Vermont, Burlington, VT
05405, USA
Author for correspondence (e-mail:
kinseys{at}uncwil.edu
)
Accepted 8 April 2002
Flux was examined through the reaction catalyzed by arginine kinase in
intact blue crab (Callinectes sapidus) muscle during simulated
changes in salinity. Isolated dark levator muscles from the swimming leg were
superfused with a saline solution that had an osmolarity equivalent to that of
the hemolymph under different salinity regimes. Animals were acclimated for 7
days to a salinity of 5, 17 or 35
, which corresponds to a hemolymph
osmolarity of 640, 720 or 960 mosmoll-1, respectively. Experiments
were conducted under control conditions, in which the osmolarity of the
superfusion medium matched that of the acclimated hemolymph, as well as under
hypo- and hyperosmotic conditions. These latter treatments were meant to
simulate a rapid change in environmental salinity. Pseudo-first-order
unidirectional rate constants and flux rates were measured for arginine kinase
in the forward and reverse directions using a 31P-nuclear magnetic
resonance saturation transfer method. There were no differences in the rate
constants or flux rates among the controls, indicating that arginine kinase
function is not modulated by salinity if the animal has had sufficient
acclimation time. However, the rate constants and flux rates of arginine
kinase varied over a modest 1.7-fold range across the three types of osmotic
treatments, although the range for the flux data was reduced when cell volume
changes were taken into account. The hyperosmotic treatments led to a
reduction in arginine kinase flux, while the hypo-osmotic treatments led to an
enhanced arginine kinase flux. We propose that this effect is mediated by an
increase in the concentration of perturbing inorganic ions under hyperosmotic
conditions and a decrease in the concentration of such ions during the
hypo-osmotic treatments.
Key words: arginine kinase, osmoregulation, haemolymph, muscle, blue crab, Callinectes sapidus, crustacean, nuclear magnetic resonance, salinity
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