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Endogenous lactate transport in Xenopus laevis oocyte: dependence on cytoskeleton and regulation by protein kinases
Authors:Marisa Tosco  Alide Faelli  Giulia Gastaldi  Markus Paulmichl  Maria Novella Orsenigo
Affiliation:(1) Dipartimento di Scienze Biomolecolari e Biotecnologie, Università di Milano, via Celoria 26, 20133 Milano, Italy;(2) Dipartimento di Medicina Sperimentale, Università di Pavia, via Forlanini 6, 27100 Pavia, Italy
Abstract:Carbon flux in Xenopus laevis oocyte is glycogenic and an endogenous monocarboxylate transporter is responsible for intracellular lactate uptake. The aim of the present study was to determine if direct activation of protein kinases C and A modulates the activity of lactate transporter, as well as to investigate the possible role of cytoskeleton in these regulatory phenomena. The modulation was studied in isolated Xenopus oocytes of stage V–VI by measuring 14C-lactate uptake, both in the absence and in the presence of cytoskeletal-perturbing toxins. We found that the basal lactate transporter activity depends on the integrity of the cytoskeleton since it is partially inhibited by cytoskeleton disorganisation. Both PKA and PKC activation caused a significant decrease in transport activity and this decrease could be blocked by specific protein kinase inhibitors. The evidenced effects were not additive. Transport inhibition was annulled by agents that destabilize actin filaments or microtubules. We conclude that both protein kinases A and C, whose effects are mediated by cytoskeleton, negatively regulate the endogenous lactate transporter of Xenopus oocyte, suggesting that these kinases may have a role in the control of cytosolic pyruvate/lactate pool in the oocyte. All the experiments in this study comply with the current laws of Italy.
Keywords:Xenopus oocyte  Lactate transport  PKA  PKC  Cytoskeleton
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