GluN2B but not GluN2A for basal dendritic growth of cortical pyramidal neurons
- NMDA receptors are important players for neuronal differentiation. We previously reported that antagonizing NMDA receptors with APV blocked the growth-promoting effects evoked by the overexpression of specific calcium-permeable or flip-spliced AMPA receptor subunits and of type I transmembrane AMPA receptor regulatory proteins which both exclusively modify apical dendritic length and branching of cortical pyramidal neurons. These findings led us to characterize the role of GluN2B and GluN2A for dendritogenesis using organotypic cultures of rat visual cortex. Antagonizing GluN2B with ifenprodil and Ro25-6981 strongly impaired basal dendritic growth of supra- and infragranular pyramidal cells at DIV 5–10, but no longer at DIV 15–20. Growth recovered after washout, and protein blots revealed an increase of synaptic GluN2B-containing receptors as indicated by a enhanced phosphorylation of the tyrosine 1472 residue. Antagonizing GluN2A with TCN201 and NVP-AAM077 was ineffective at both ages. Dendrite growth of non-pyramidal interneurons was not altered. We attempted to overexpress GluN2A and GluN2B. However, although the constructs delivered currents in HEK cells, there were neither effects on dendrite morphology nor an enhanced sensitivity to NMDA. Further, co-expressing GluN1-1a and GluN2B did not alter dendritic growth. Visualization of overexpressed, tagged GluN2 proteins was successful after immunofluorescence for the tag which delivered rather weak staining in HEK cells as well as in neurons. This suggested that the level of overexpression is too weak to modify dendrite growth. In summary, endogenous GluN2B, but not GluN2A is important for pyramidal cell basal dendritic growth during an early postnatal time window.
Author: | Steffen GondaORCiDGND, Jan GiesenGND, Alexander SieberathGND, Fabian WestGND, Raoul BuchholzGND, Oliver KlattGND, Tim ZiebarthGND, Andrea RäkGND, Sabine KleinhubbertGND, Christian RiedelORCiDGND, Michael HollmannORCiDGND, Mohammad HamadGND, Andreas ReinerORCiDGND, Petra WahleORCiDGND |
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URN: | urn:nbn:de:hbz:294-78838 |
DOI: | https://doi.org/10.3389/fnana.2020.571351 |
Parent Title (English): | Frontiers in neuroanatomy |
Publisher: | Frontiers Media |
Place of publication: | Lausanne |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2021/02/17 |
Date of first Publication: | 2020/11/13 |
Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
Tag: | Open Access Fonds GluN1; ampakine CX546; biolistic transfection; inhibition; rodent neocortex |
Volume: | 14 |
Issue: | Artikel 571351 |
First Page: | 571351-1 |
Last Page: | 571351-18 |
Note: | Article Processing Charge funded by the Deutsche Forschungsgemeinschaft (DFG) and the Open Access Publication Fund of Ruhr-Universität Bochum. |
Institutes/Facilities: | Lehrstuhl Biochemie I, Rezeptorbiochemie |
Lehrstuhl für Allgemeine Zoologie und Neurobiologie, Arbeitsgruppe für Entwicklungsneurobiologie | |
Dewey Decimal Classification: | Naturwissenschaften und Mathematik / Biowissenschaften, Biologie, Biochemie |
faculties: | Fakultät für Chemie und Biochemie |
Fakultät für Biologie und Biotechnologie | |
Licence (English): | Creative Commons - CC BY 4.0 - Attribution 4.0 International |