Functional metabolic interactions of human neuron-astrocyte 3D in vitro networks.
The generation of human neural tissue-like 3D structures holds great promise for disease modeling, drug discovery and regenerative medicine strategies. Promoting the establishment of complex cell-cell interactions, 3D culture systems enable the development of human cell-based models with increased physiological relevance, over monolayer cultures. Here, we demonstrate the establishment of neuronal and astrocytic metabolic signatures and shuttles in a human 3D neural cell model, namely the glutamine-glutamate-GABA shuttle. This was indicated by labeling of neuronal GABA following incubation with the glia-specific substrate [2-(13)C]acetate, which decreased by methionine sulfoximine-induced inhibition of the glial enzyme glutamine synthetase. Cell metabolic specialization was further demonstrated by higher pyruvate carboxylase-derived labeling in glutamine than in glutamate, indicating its activity in astrocytes and not in neurons. Exposure to the neurotoxin acrylamide resulted in intracellular accumulation of glutamate and decreased GABA synthesis. These results suggest an acrylamide-induced impairment of neuronal synaptic vesicle trafficking and imbalanced glutamine-glutamate-GABA cycle, due to loss of cell-cell contacts at synaptic sites. This work demonstrates, for the first time to our knowledge, that neural differentiation of human cells in a 3D setting recapitulates neuronal-astrocytic metabolic interactions, highlighting the relevance of these models for toxicology and better understanding the crosstalk between human neural cells.
iBET, Instituto de Biologia Experimental e Biológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Daniel
D
Simão
iBET, Instituto de Biologia Experimental e Biológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Ana P
AP
Terrasso
iBET, Instituto de Biologia Experimental e Biológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Ana P
AP
Teixeira
iBET, Instituto de Biologia Experimental e Biológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Catarina
C
Brito
Department of Neuroscience, Norwegian University of Science and Technology (NTNU), Trondheim, Norway.|Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, 2100, Denmark.
Ursula
U
Sonnewald
iBET, Instituto de Biologia Experimental e Biológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Paula M
PM
Alves
D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D001921:Brain / Q000000981:diagnostic imaging / Q000378:metabolism*; D002247:Carbon Isotopes / Q000737:chemistry; D002450:Cell Communication; D005974:Glutamate-Ammonia Ligase / Q000235:genetics; D018698:Glutamic Acid / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D009415:Nerve Net / Q000166:cytology / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000096:biosynthesis / Q000378:metabolism
D002247:Carbon Isotopes; D005973:Glutamine; D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid; D005974:Glutamate-Ammonia Ligase