Defined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neurons
Abstract Background Mutations in the gene DISC1 are associated with increased risk for schizophrenia, bipolar disorder and major depression. The study of mutated DISC1 represents a well-known and comprehensively characterized approach to understand neuropsychiatric disease mechanisms. However, previ...
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | BMC Neuroscience |
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| Online Access: | https://doi.org/10.1186/s12868-024-00858-z |
| _version_ | 1797275729443946496 |
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| author | Johanna Heider Aaron Stahl Denise Sperlich Sophia-Marie Hartmann Sabrina Vogel Ricarda Breitmeyer Markus Templin Hansjürgen Volkmer |
| author_facet | Johanna Heider Aaron Stahl Denise Sperlich Sophia-Marie Hartmann Sabrina Vogel Ricarda Breitmeyer Markus Templin Hansjürgen Volkmer |
| author_sort | Johanna Heider |
| collection | DOAJ |
| description | Abstract Background Mutations in the gene DISC1 are associated with increased risk for schizophrenia, bipolar disorder and major depression. The study of mutated DISC1 represents a well-known and comprehensively characterized approach to understand neuropsychiatric disease mechanisms. However, previous studies have mainly used animal models or rather heterogeneous populations of iPSC-derived neurons, generated by undirected differentiation, to study the effects of DISC1 disruption. Since major hypotheses to explain neurodevelopmental, psychiatric disorders rely on altered neuronal connectivity observed in patients, an ideal iPSC-based model requires accurate representation of the structure and complexity of neuronal circuitries. In this study, we made use of an isogenic cell line with a mutation in DISC1 to study neuronal synaptic phenotypes in a culture system comprising a defined ratio of NGN2 and ASCL1/DLX2 (AD2)-transduced neurons, enriched for glutamatergic and GABAergic neurons, respectively, to mimic properties of the cortical microcircuitry. Results In heterozygous DISC1 mutant neurons, we replicated the expected phenotypes including altered neural progenitor proliferation as well as neurite outgrowth, deregulated DISC1-associated signaling pathways, and reduced synaptic densities in cultures composed of glutamatergic neurons. Cultures comprising a defined ratio of NGN2 and AD2 neurons then revealed considerably increased GABAergic synapse densities, which have not been observed in any iPSC-derived model so far. Increased inhibitory synapse densities could be associated with an increased efficiency of GABAergic differentiation, which we observed in AD2-transduced cultures of mutant neurons. Additionally, we found increased neuronal activity in GABAergic neurons through calcium imaging while the activity pattern of glutamatergic neurons remained unchanged. Conclusions In conclusion, our results demonstrate phenotypic differences in a co-culture comprising a defined ratio of DISC1 mutant NGN2 and AD2 neurons, as compared to culture models comprising only one neuronal cell type. Altered synapse numbers and neuronal activity imply that DISC1 impacts the excitatory/inhibitory balance in NGN2/AD2 co-cultures, mainly through increased GABAergic input. |
| first_indexed | 2024-03-07T15:17:41Z |
| format | Article |
| id | doaj.art-1113a8edc4e34a2a87fef4ae3a9fb152 |
| institution | Directory Open Access Journal |
| issn | 1471-2202 |
| language | English |
| last_indexed | 2024-03-07T15:17:41Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
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| series | BMC Neuroscience |
| spelling | doaj.art-1113a8edc4e34a2a87fef4ae3a9fb1522024-03-05T17:47:44ZengBMCBMC Neuroscience1471-22022024-03-0125112010.1186/s12868-024-00858-zDefined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neuronsJohanna Heider0Aaron Stahl1Denise Sperlich2Sophia-Marie Hartmann3Sabrina Vogel4Ricarda Breitmeyer5Markus Templin6Hansjürgen Volkmer7Department of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenDepartment of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenDepartment of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenDepartment of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenDepartment of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenDepartment of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenDepartment of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenDepartment of Pharma and Biotech, NMI Natural and Medical Sciences Institute at the University of TübingenAbstract Background Mutations in the gene DISC1 are associated with increased risk for schizophrenia, bipolar disorder and major depression. The study of mutated DISC1 represents a well-known and comprehensively characterized approach to understand neuropsychiatric disease mechanisms. However, previous studies have mainly used animal models or rather heterogeneous populations of iPSC-derived neurons, generated by undirected differentiation, to study the effects of DISC1 disruption. Since major hypotheses to explain neurodevelopmental, psychiatric disorders rely on altered neuronal connectivity observed in patients, an ideal iPSC-based model requires accurate representation of the structure and complexity of neuronal circuitries. In this study, we made use of an isogenic cell line with a mutation in DISC1 to study neuronal synaptic phenotypes in a culture system comprising a defined ratio of NGN2 and ASCL1/DLX2 (AD2)-transduced neurons, enriched for glutamatergic and GABAergic neurons, respectively, to mimic properties of the cortical microcircuitry. Results In heterozygous DISC1 mutant neurons, we replicated the expected phenotypes including altered neural progenitor proliferation as well as neurite outgrowth, deregulated DISC1-associated signaling pathways, and reduced synaptic densities in cultures composed of glutamatergic neurons. Cultures comprising a defined ratio of NGN2 and AD2 neurons then revealed considerably increased GABAergic synapse densities, which have not been observed in any iPSC-derived model so far. Increased inhibitory synapse densities could be associated with an increased efficiency of GABAergic differentiation, which we observed in AD2-transduced cultures of mutant neurons. Additionally, we found increased neuronal activity in GABAergic neurons through calcium imaging while the activity pattern of glutamatergic neurons remained unchanged. Conclusions In conclusion, our results demonstrate phenotypic differences in a co-culture comprising a defined ratio of DISC1 mutant NGN2 and AD2 neurons, as compared to culture models comprising only one neuronal cell type. Altered synapse numbers and neuronal activity imply that DISC1 impacts the excitatory/inhibitory balance in NGN2/AD2 co-cultures, mainly through increased GABAergic input.https://doi.org/10.1186/s12868-024-00858-zDISC1E/I imbalanceGABAergic neuronsGlutamatergic neuronsiPSCCo-culture |
| spellingShingle | Johanna Heider Aaron Stahl Denise Sperlich Sophia-Marie Hartmann Sabrina Vogel Ricarda Breitmeyer Markus Templin Hansjürgen Volkmer Defined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neurons BMC Neuroscience DISC1 E/I imbalance GABAergic neurons Glutamatergic neurons iPSC Co-culture |
| title | Defined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neurons |
| title_full | Defined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neurons |
| title_fullStr | Defined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neurons |
| title_full_unstemmed | Defined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neurons |
| title_short | Defined co-cultures of glutamatergic and GABAergic neurons with a mutation in DISC1 reveal aberrant phenotypes in GABAergic neurons |
| title_sort | defined co cultures of glutamatergic and gabaergic neurons with a mutation in disc1 reveal aberrant phenotypes in gabaergic neurons |
| topic | DISC1 E/I imbalance GABAergic neurons Glutamatergic neurons iPSC Co-culture |
| url | https://doi.org/10.1186/s12868-024-00858-z |
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