Disabling VEGF-response of Purkinje Cells by downregulation of \(\it KDR\) via miRNA-204-5p
- The vascular endothelial growth factor (VEGF) is well known for its wide-ranging functions, not only in the vascular system, but also in the central (CNS) and peripheral nervous system (PNS). To study the role of VEGF in neuronal protection, growth and maturation processes have recently attracted much interest. These effects are mainly mediated by VEGF receptor 2 (VEGFR-2). Current studies have shown the age-dependent expression of VEGFR-2 in Purkinje cells (PC), promoting dendritogenesis in neonatal, but not in mature stages. We hypothesize that microRNAs (miRNA/miR) might be involved in the regulation of VEGFR-2 expression during the development of PC. In preliminary studies, we performed a miRNA profiling and identified miR204-5p as a potential regulator of VEGFR-2 expression. In the recent study, organotypic slice cultures of rat cerebella (postnatal day (p) 1 and 9) were cultivated and VEGFR-2 expression in PC was verified via immunohistochemistry. Additionally, PC at age p9 and p30 were isolated from cryosections by laser microdissection (LMD) to analyse VEGFR-2 expression by quantitative RT-PCR. To investigate the influence of miR204-5p on VEGFR-2 levels in PC, synthetic constructs including short hairpin (sh)-miR204-5p cassettes (miRNA-mimics), were microinjected into PC. The effects were analysed by confocal laser scanning microscopy (CLSM) and morphometric analysis. For the first time, we could show that miR204-5p has a negative effect on VEGF sensitivity in juvenile PC, resulting in a significant decrease of dendritic growth compared to untreated juvenile PC. In mature PC, the overexpression of miR204-5p leads to a shrinkage of dendrites despite VEGF treatment. The results of this study illustrate, for the first time, which miR204-5p expression has the potential to play a key role in cerebellar development by inhibiting VEGFR-2 expression in PC.
Author: | Julian GehmeyrGND, Abdelouahid MaghnoujGND, Jonas TjadenGND, Matthias VorgerdORCiDGND, Stephan HahnORCiDGND, Veronika MatschkeORCiDGND, Verena TheisGND, Carsten TheißORCiDGND |
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URN: | urn:nbn:de:hbz:294-85743 |
DOI: | https://doi.org/10.3390/ijms22042173 |
Parent Title (German): | International journal of molecular sciences |
Publisher: | MDPI |
Place of publication: | Basel |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2022/02/10 |
Date of first Publication: | 2021/02/22 |
Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
Tag: | Open Access Fonds KDR; Purkinje cells; VEGF; VEGFR-2; cerebellum; dendritic growth; miRNA-204-5p; microinjection; organotypic slice cultures |
Volume: | 22 |
Issue: | 4, Artikel 2173 |
First Page: | 2173-1 |
Last Page: | 2173-22 |
Note: | Article Processing Charge funded by the Open Access Publication Fund of Ruhr-Universität Bochum. |
Institutes/Facilities: | Zentrum für Klinische Forschung Bochum |
Berufsgenossenschaftliches Universitätsklinikum Bergmannsheil, Neurologische Universitätsklinik und Poliklinik | |
Institut für Anatomie, Abteilung für Cytologie | |
Institut für Molekulare Gastroenterologische Onkologie | |
Muskelzentrum Ruhrgebiet | |
open_access (DINI-Set): | open_access |
Licence (English): | Creative Commons - CC BY 4.0 - Attribution 4.0 International |