Adipogranular cells from the mantle tissue of Mytilus edulis L. I. Isolation, purification and biochemical characteristics of dispersed cells

Abstract

High-density adipogranular (ADG) cells were isolated from the mantle tissue of female mussels Mytilus edulis L. by enzymic dispersion and centrifugation through Percoll of density 1.08 g/ml. The cells were viable as judged by microscopic examination, exclusion of Trypan Blue, nonleakage of intracellular glutamate-oxaloacetate transaminase (GOT) and malate dehydrogenase (MDH), and utilization of exogenous [2- 3H] and [U- 14C] glucose. In dual-labelled experiments, the levels of [ 3H]glucose and [ 14C]glucose in cellular glycogen were equivalent to the utilization of 80 nmol glucose/h/g wet wt and 120 nmol glucose/h/g wet wt, respectively, at an external glucose concentration of 0.5 mM. The rate of 14CO 2 production was very low (18 nmol glucose/h/g wet wt), representing 6% of the total glucose utilization as measured by detritiation of [2- 3H] glucose. Recovery of 3H 2O from [2- 3H] glucose was linear with time over a 3-h incubation and showed saturation kinetics for glucose utilization, with an apparent K m of 0.56 ± 0.09 mM and a V max of 632 ± 48 nmol glucose/h/g wet wt.