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A new mouse line reporting the translation of brain-derived neurotrophic factor using green fluorescent protein

Wosnitzka, Erin, Nan, Xinsheng, Nan, Jeff, Chacón-Fernández, Pedro, Kussmaul, Lothar, Schuler, Michael, Hengerer, Bastian and Barde, Yves-Alain 2020. A new mouse line reporting the translation of brain-derived neurotrophic factor using green fluorescent protein. eNeuro 7 (1) , ENEURO.0462-19.2019. 10.1523/ENEURO.0462-19.2019

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Abstract

While BDNF is receiving considerable attention for its role in synaptic plasticity and in nervous system dysfunction, identifying brain circuits involving BDNF-expressing neurons has been challenging. BDNF levels are very low in most brain areas, except for the large mossy fiber terminals in the hippocampus where BDNF accumulates at readily detectable levels. This report describes the generation of a mouse line allowing the detection of single brain cells synthesizing BDNF. A bicistronic construct encoding BDNF tagged with a P2A sequence preceding GFP allows the translation of BDNF and GFP as separate proteins. Following its validation with transfected cells, this construct was used to replace the endogenous Bdnf gene. Viable and fertile homozygote animals were generated, with the GFP signal marking neuronal cell bodies translating the Bdnf mRNA. Importantly, the distribution of immunoreactive BDNF remained unchanged, as exemplified by its accumulation in mossy fiber terminals in the transgenic animals. GFP-labeled neurons could be readily visualized in distinct layers in the cerebral cortex where BDNF has been difficult to detect with currently available reagents. In the hippocampal formation, quantification of the GFP signal revealed that <10% of the neurons do not translate the Bdnf mRNA at detectable levels, with the highest proportion of strongly labeled neurons found in CA3.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Additional Information: This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.
Publisher: Society for Neuroscience: eNeuro
ISSN: 2373-2822
Date of First Compliant Deposit: 23 January 2020
Date of Acceptance: 15 December 2019
Last Modified: 09 Nov 2020 14:00
URI: http://orca-mwe.cf.ac.uk/id/eprint/128953

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