Characterization hiPSC-derived neural progenitor cells and neurons to investigate the role of NOS1AP isoforms in human neuron dendritogenesis
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 prefro...
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| Published in | Molecular and cellular neuroscience Vol. 109; p. 103562 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
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Elsevier Inc
01.12.2020
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| Abstract | 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.
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•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. |
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| AbstractList | 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. 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. 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.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. 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. |
| ArticleNumber | 103562 |
| Author | Bhattiprolu, Atul K. Paradiso, Kenneth G. Hernandez, Kristina Firestein, Bonnie L. Dudzinski, Natasha R. Crosta, Christen M. Moore, Jennifer C. Clarke, Stephen G. Brzustowicz, Linda M. Fu, Allen Y. |
| AuthorAffiliation | 2 Neurosciences Graduate Program, Rutgers, The State University of New Jersey, Piscataway, New Jersey 4 Department of Genetics, Rutgers, The State University of New Jersey, 604 Allison Road, Piscataway, NJ, USA 08854-8082 1 Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, New Jersey 3 Molecular Biosciences Graduate Program, Rutgers, The State University of New Jersey, Piscataway, New Jersey |
| AuthorAffiliation_xml | – name: 3 Molecular Biosciences Graduate Program, Rutgers, The State University of New Jersey, Piscataway, New Jersey – name: 1 Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, New Jersey – name: 2 Neurosciences Graduate Program, Rutgers, The State University of New Jersey, Piscataway, New Jersey – name: 4 Department of Genetics, Rutgers, The State University of New Jersey, 604 Allison Road, Piscataway, NJ, USA 08854-8082 |
| Author_xml | – sequence: 1 givenname: Christen M. surname: Crosta fullname: Crosta, Christen M. organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA – sequence: 2 givenname: Kristina surname: Hernandez fullname: Hernandez, Kristina organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA – sequence: 3 givenname: Atul K. surname: Bhattiprolu fullname: Bhattiprolu, Atul K. organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA – sequence: 4 givenname: Allen Y. surname: Fu fullname: Fu, Allen Y. organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA – sequence: 5 givenname: Jennifer C. surname: Moore fullname: Moore, Jennifer C. organization: Department of Genetics, Rutgers, The State University of New Jersey, 604 Allison Road, Piscataway, NJ 08854-8082, USA – sequence: 6 givenname: Stephen G. surname: Clarke fullname: Clarke, Stephen G. organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA – sequence: 7 givenname: Natasha R. surname: Dudzinski fullname: Dudzinski, Natasha R. organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA – sequence: 8 givenname: Linda M. surname: Brzustowicz fullname: Brzustowicz, Linda M. organization: Department of Genetics, Rutgers, The State University of New Jersey, 604 Allison Road, Piscataway, NJ 08854-8082, USA – sequence: 9 givenname: Kenneth G. surname: Paradiso fullname: Paradiso, Kenneth G. organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA – sequence: 10 givenname: Bonnie L. surname: Firestein fullname: Firestein, Bonnie L. email: firestein@biology.rutgers.edu organization: Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA |
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| Keywords | hiPSC-derived neurons Arborization d-Serine Morphology NOS1AP Antipsychotic medication |
| Language | English |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Christen M. Crosta: Data curation; Formal analysis; Validation; Visualization; Writing - review & editing; Kristina Hernandez: Data curation; Formal analysis; Investigation; Methodology; Visualization; Writing - original draft; Atul K. Bhattiprolu: Data curation; Formal analysis; Validation; Allen Y. Fu: Data curation; Formal analysis; Jennifer C. Moore: Conceptualization; Methodology; Writing - review & editing; Stephen G. Clarke: Data curation; Formal analysis; Investigation; Natasha R. Dudzinski: Data curation; Formal analysis; Investigation; Linda M. Brzustowicz: Conceptualization; Resources; Kenneth G. Paradiso: Conceptualization; Formal analysis; Methodology; Visualization; Writing - review & editing; Bonnie L. Firestein: Conceptualization; Funding acquisition; Methodology; Project administration; Resources; Supervision; Visualization; Writing - original draft; Writing - review & editing. Authorship Statements |
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| SubjectTerms | 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 |
| Title | Characterization hiPSC-derived neural progenitor cells and neurons to investigate the role of NOS1AP isoforms in human neuron dendritogenesis |
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