Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation
Denervation of neurons is a network consequence of brain injury. The effects of denervation on neurons can be readily studied in vitro using organotypic slice cultures of entorhinal cortex and hippocampus. Following transection of the entorhino-dentate projection, granule cells (GCs) are denervated...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Neuroanatomy |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fnana.2024.1513511/full |
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| author | Davide Greco Alexander Drakew Nina Rößler Nina Rößler Tassilo Jungenitz Peter Jedlicka Peter Jedlicka Thomas Deller |
| author_facet | Davide Greco Alexander Drakew Nina Rößler Nina Rößler Tassilo Jungenitz Peter Jedlicka Peter Jedlicka Thomas Deller |
| author_sort | Davide Greco |
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| description | Denervation of neurons is a network consequence of brain injury. The effects of denervation on neurons can be readily studied in vitro using organotypic slice cultures of entorhinal cortex and hippocampus. Following transection of the entorhino-dentate projection, granule cells (GCs) are denervated and show on average a transient loss of spines on their denervated distal dendrites but not on their non-denervated proximal dendrites. In the present study, we addressed the question how single GCs and their denervated and non-denervated segments react to entorhinal denervation. Local adeno-associated virus (AAV)-injections were employed to transduce dentate GCs with tdTomato and entorhinal projection neurons with EGFP. This made it possible to visualize both innervating entorhinal fibers and their target neurons and to identify dendritic segments located in the “entorhinal” and the “hippocampal” zone of the dentate gyrus. Confocal time-lapse imaging was used to image distal and proximal segments of single GCs after entorhinal denervation. Time-matched non-denervated cultures served as controls. In line with previous reports, average dendritic spine loss was ~30% (2–4 days post-lesion) in the denervated zone. However, individual GCs showed considerable variability in their response to denervation in both layers, and both decreases as well as increases in spine density were observed at the single cell level. Based on the standard deviations and the effect sizes observed in this study, a computer simulation yielded recommendations for the minimum number of neurons that should be analyzed in future studies using the entorhinal in vitro denervation model. |
| format | Article |
| id | doaj-art-245835684f1842f2a11536d166324574 |
| institution | Kabale University |
| issn | 1662-5129 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Neuroanatomy |
| spelling | doaj-art-245835684f1842f2a11536d1663245742025-01-21T08:36:58ZengFrontiers Media S.A.Frontiers in Neuroanatomy1662-51292025-01-011810.3389/fnana.2024.15135111513511Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervationDavide Greco0Alexander Drakew1Nina Rößler2Nina Rößler3Tassilo Jungenitz4Peter Jedlicka5Peter Jedlicka6Thomas Deller7Institute for Clinical Neuroanatomy, Faculty of Medicine, Goethe-University, Frankfurt, GermanyInstitute for Clinical Neuroanatomy, Faculty of Medicine, Goethe-University, Frankfurt, GermanyInstitute for Clinical Neuroanatomy, Faculty of Medicine, Goethe-University, Frankfurt, Germany3R Computer-Based Modelling, Faculty of Medicine, ICAR3R, Justus-Liebig-University, Giessen, GermanyInstitute for Clinical Neuroanatomy, Faculty of Medicine, Goethe-University, Frankfurt, GermanyInstitute for Clinical Neuroanatomy, Faculty of Medicine, Goethe-University, Frankfurt, Germany3R Computer-Based Modelling, Faculty of Medicine, ICAR3R, Justus-Liebig-University, Giessen, GermanyInstitute for Clinical Neuroanatomy, Faculty of Medicine, Goethe-University, Frankfurt, GermanyDenervation of neurons is a network consequence of brain injury. The effects of denervation on neurons can be readily studied in vitro using organotypic slice cultures of entorhinal cortex and hippocampus. Following transection of the entorhino-dentate projection, granule cells (GCs) are denervated and show on average a transient loss of spines on their denervated distal dendrites but not on their non-denervated proximal dendrites. In the present study, we addressed the question how single GCs and their denervated and non-denervated segments react to entorhinal denervation. Local adeno-associated virus (AAV)-injections were employed to transduce dentate GCs with tdTomato and entorhinal projection neurons with EGFP. This made it possible to visualize both innervating entorhinal fibers and their target neurons and to identify dendritic segments located in the “entorhinal” and the “hippocampal” zone of the dentate gyrus. Confocal time-lapse imaging was used to image distal and proximal segments of single GCs after entorhinal denervation. Time-matched non-denervated cultures served as controls. In line with previous reports, average dendritic spine loss was ~30% (2–4 days post-lesion) in the denervated zone. However, individual GCs showed considerable variability in their response to denervation in both layers, and both decreases as well as increases in spine density were observed at the single cell level. Based on the standard deviations and the effect sizes observed in this study, a computer simulation yielded recommendations for the minimum number of neurons that should be analyzed in future studies using the entorhinal in vitro denervation model.https://www.frontiersin.org/articles/10.3389/fnana.2024.1513511/fullhippocampal slice culturesregenerationperforant pathaxotomydeafferentationplasticity |
| spellingShingle | Davide Greco Alexander Drakew Nina Rößler Nina Rößler Tassilo Jungenitz Peter Jedlicka Peter Jedlicka Thomas Deller Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation Frontiers in Neuroanatomy hippocampal slice cultures regeneration perforant path axotomy deafferentation plasticity |
| title | Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation |
| title_full | Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation |
| title_fullStr | Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation |
| title_full_unstemmed | Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation |
| title_short | Time-lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation |
| title_sort | time lapse imaging of identified granule cells in the mouse dentate gyrus after entorhinal lesion in vitro reveals heterogeneous cellular responses to denervation |
| topic | hippocampal slice cultures regeneration perforant path axotomy deafferentation plasticity |
| url | https://www.frontiersin.org/articles/10.3389/fnana.2024.1513511/full |
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