Characterization hiPSC-derived neural progenitor cells and neurons to investigate the role of NOS1AP isoforms in human neuron dendritogenesis

• Published in: Molecular and cellular neuroscience Vol. 109; p. 103562
• Main Authors: Crosta, Christen M., Hernandez, Kristina, Bhattiprolu, Atul K., Fu, Allen Y., Moore, Jennifer C., Clarke, Stephen G., Dudzinski, Natasha R., Brzustowicz, Linda M., Paradiso, Kenneth G., Firestein, Bonnie L.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2020
• Subjects: Adaptor Proteins, Signal Transducing - biosynthesis; Adaptor Proteins, Signal Transducing - genetics; Adaptor Proteins, Signal Transducing - physiology; Antipsychotic medication; Arborization; Cells, Cultured; Clozapine - pharmacology; d-Serine; Dendrites - ultrastructure; Drug Evaluation, Preclinical; Fluphenazine - pharmacology; Gene Expression Regulation - drug effects; Glutamic Acid - physiology; Haloperidol - pharmacology; hiPSC-derived neurons; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Ion Channels - physiology; Morphology; Nerve Tissue Proteins - physiology; Neural Stem Cells - cytology; Neural Stem Cells - metabolism; Neurons - cytology; Neurons - drug effects; Neurons - metabolism; NOS1AP; Oligopeptides - pharmacology; Patch-Clamp Techniques; Protein Isoforms - physiology; Schizophrenia - etiology; Schizophrenia - genetics; Serine - pharmacology; hiPSC-derived neurons; Arborization; d-Serine; Morphology; NOS1AP; Antipsychotic medication
• ISSN: 1044-7431; 1095-9327; 1095-9327
• DOI: 10.1016/j.mcn.2020.103562

Abnormal dendritic arbor development has been implicated in a number of neurodevelopmental disorders, such as autism and Rett syndrome, and the neuropsychiatric disorder schizophrenia. Postmortem brain samples from subjects with schizophrenia show elevated levels of NOS1AP in the dorsolateral prefrontal cortex, a region of the brain associated with cognitive function. We previously reported that the long isoform of NOS1AP (NOS1AP-L), but not the short isoform (NOS1AP-S), negatively regulates dendrite branching in rat hippocampal neurons. To investigate the role that NOS1AP isoforms play in human dendritic arbor development, we adapted methods to generate human neural progenitor cells and neurons using induced pluripotent stem cell (iPSC) technology. We found that increased protein levels of either NOS1AP-L or NOS1AP-S decrease dendrite branching in human neurons at the developmental time point when primary and secondary branching actively occurs. Next, we tested whether pharmacological agents can decrease the expression of NOS1AP isoforms. Treatment of human iPSC-derived neurons with d-serine, but not clozapine, haloperidol, fluphenazine, or GLYX-13, results in a reduction in endogenous NOS1AP-L, but not NOS1AP-S, protein expression; however, d-serine treatment does not reverse decreases in dendrite number mediated by overexpression of NOS1AP isoforms. In summary, we demonstrate how an in vitro model of human neuronal development can help in understanding the etiology of schizophrenia and can also be used as a platform to screen drugs for patients. [Display omitted] •Increased protein levels of NOS1AP decrease dendrite branching in human neurons in vitro.•Treatment of human neurons with d-serine reduces NOS1AP-L, but not NOS1AP-S, protein expression.•Treatment of human iPSC-derived neurons with clozapine, haloperidol, or fluphenazine, does not alter NOS1AP expression.•NOS1AP overexpression decreases dendrite branching in hiPSC-derived neurons.•d-Serine reduces NOS1AP-L but not exogenous NOS1AP-promoted decreased dendrites.