RESUMO
Photoreceptor ribbon synapses tonically release glutamate. To ensure efficient signal transmission and prevent glutamate toxicity, a highly efficient glutamate removal system provided by members of the SLC1 gene family is required. By using a combination of biophysical and in vivo studies, we elucidate the role of excitatory amino acid transporter 2 (EAAT2) proteins in synaptic glutamate homeostasis at the zebrafish photoreceptor synapse. The main glutamate sink is provided by the glial EAAT2a, reflected by reduced electroretinographic responses in EAAT2a-depleted larvae. EAAT2b is located on the tips of cone pedicles and contributes little to glutamate reuptake. However, this transporter displays both a large chloride conductance and leak current, being important in stabilizing the cone resting potential. This work demonstrates not only how proteins originating from the same gene family can complement each other's expression profiles and biophysical properties, but also how presynaptic and glial transporters are coordinated to ensure efficient synaptic transmission at glutamatergic synapses of the central nervous system.
Assuntos
Transportador 2 de Aminoácido Excitatório/metabolismo , Células Fotorreceptoras/classificação , Células Fotorreceptoras/metabolismo , Terminações Pré-Sinápticas/metabolismo , Animais , Animais Geneticamente Modificados , Transportador 2 de Aminoácido Excitatório/genética , Olho/citologia , Regulação da Expressão Gênica/genética , Ácido Glutâmico/metabolismo , Técnicas In Vitro , Larva , Morfolinos/genética , Morfolinos/metabolismo , Oócitos/fisiologia , Técnicas de Patch-Clamp , Retina/anatomia & histologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Sinapses , Transmissão Sináptica/fisiologia , Vias Visuais/fisiologia , Xenopus , Peixe-ZebraRESUMO
The formation of functional neuronal circuits relies on accurate migration and proper axonal outgrowth of neuronal precursors. On the route to their targets migrating cells and growing axons depend on both, directional information from neurotropic cues and adhesive interactions mediated via extracellular matrix molecules or neighbouring cells. The inactivation of guidance cues or the interference with cell adhesion can cause severe defects in neuronal migration and axon guidance. In this study we have analyzed the function of the MAM domain containing glycosylphosphatidylinositol anchor 2A (MDGA2A) protein in zebrafish cranial motoneuron development. MDGA2A is prominently expressed in distinct clusters of cranial motoneurons, especially in the ones of the trigeminal and facial nerves. Analyses of MDGA2A knockdown embryos by light sheet and confocal microscopy revealed impaired migration and aberrant axonal outgrowth of these neurons; suggesting that adhesive interactions mediated by MDGA2A are required for the proper arrangement and outgrowth of cranial motoneuron subtypes.
