Our conclusion is that astrocytes are the key APC with respect to adaptive responses to GFP expression. This, of course, is not likely to be a problem when self-proteins are expressed, but in the present study we observed, as previously, the activation of Iba1 and upregulation of MHC-II on astrocytes and microglia. In one embodiment, the toxicity, e.g., in vivo acute neurotoxicity, of the molecule is tested by measuring intracellular free calcium oscillations (calcium oscillations) in vitro in neuronal cells which are in contact or have been in contact with the molecule. In certain embodiments, the present disclosure provides a method of identifying or determining a molecule having intolerable in vivo neurotoxicity comprising measuring calcium oscillations in vitro in neuronal cells after being in contact with the molecule in the neuronal cells. In some embodiments, the calcium oscillations for a molecule having tolerable in vivo acute neurotoxicity are compared with the calcium oscillations in a cell not exposed to the molecule.

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