Abstract
Neurons release neurotransmitters such as glutamate to communicate with each other and to coordinate brain functioning. As increased glutamate release is indicative of neuronal maturation and activity, a system that can measure glutamate levels over time within the same tissue and/or culture system is highly advantageous for neurodevelopmental investigation. To address such challenges, we develop for the first time a convenient method to realize functionalized borosilicate glass capillaries with nanostructured texture as an electrochemical biosensor to detect glutamate release from cerebral organoids generated from human embryonic stem cells (hESC) that mimic various brain regions. The biosensor shows a clear catalytic activity toward the oxidation of glutamate with a sensitivity of 93 ± 9.5 nA·µM−1·cm−2. It was found that the enzyme-modified microelectrodes can detect glutamate in a wide linear range from 5 µM to 0.5 mM with a limit of detection (LOD) down to 5.6 ± 0.2 µM. Measurements were performed within the organoids at different time points and consistent results were obtained. This data demonstrates the reliability of the biosensor as well as its usefulness in measuring glutamate levels across time within the same culture system.
Highlights
Glutamate is the principal excitatory neurotransmitter in the brain
(Leu); lysine (Lys) and phosphate-buffered saline solution (PBS) all purchased from Sigma–Aldrich
The nanostructured microelectrode provides an electro-active surface for enzyme immobilization with enhanced confirmation, orientation and biological activity
Summary
Glutamate is the principal excitatory neurotransmitter in the brain. In addition to its neuronal signaling role, glutamate plays critical roles in cortical development, including migration and differentiation of cortical neurons. High levels of glutamate may cause excitotoxicity and neuronal death that are associated with multiple neuronal diseases, such as ischemia [1,2], Parkinson’s disease [3,4], Alzheimer’s disease [5,6], and epilepsy [7,8,9]. These important roles of glutamate have generated a strong interest in the development of devices for the detection and monitoring of this amino acid. Neurospheres and cerebral organoids derived from human pluripotent stem cells (hPSC)
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