LRP1 is critical for the surface distribution and internalization of the NR2B NMDA receptor subtype

Molecular Neurodegeneration

Claus U Pietrzik³  Ulrich Schmitt⁴  Sascha Weggen¹  Anton Roebroek²  Sabrina Meister³  Mariola Bednorz⁴  Wladislaw Maier³ 

[1] Department of Neuropathology Molecular Neuropathology, Heinrich Heine University Duesseldorf, Moorenstr. 5, Duesseldorf, 40225, Germany;Laboratory for Experimental Mouse Genetics, Center for Human Genetics KU Leuven, Herestraat 49, Leuven B-3000, Belgium;University Medical Center of the Johannes Gutenberg-University of Mainz Institute of Pathobiochemistry, Duesbergweg 6, Mainz 55099, Germany;Department of Psychiatry and Psychotherapy, University Medical Center of the Johannes Gutenberg-University of Mainz, Untere Zahlbacher Str. 8, Mainz 55131, Germany

关键词: Cell surface expression;    PSD95;    NR2B receptor subunit;    NR1;    NMDA receptor;    NPxY2 motif;    LRP1;   

Others  :  862235
DOI  :  10.1186/1750-1326-8-25

 received in 2013-01-14, accepted in 2013-07-03,  发布年份 2013

【 摘 要 】

Background

The N-methyl-D-aspartate receptors are key mediators of excitatory transmission and are implicated in many forms of synaptic plasticity. These receptors are heterotetrameres consisting of two obligatory NR1 and two regulatory subunits, usually NR2A or NR2B. The NR2B subunits are abundant in the early postnatal brain, while the NR2A/NR2B ratio increases during early postnatal development. This shift is driven by NMDA receptor activity. A functional interplay of the Low Density Lipoprotein Receptor Related Protein 1 (LRP1) NMDA receptor has already been reported. Such abilities as interaction of LRP1 with NMDA receptor subunits or its important role in tPa-mediated NMDA receptor signaling were already demonstrated. Moreover, mice harboring a conditional neuronal knock-out mutation of the entire Lrp1 gene display NMDA-associated behavioral changes. However, the exact role of LRP1 on NMDA receptor function remains still elusive.

Results

To provide a mechanistic explanation for such effects we investigated whether an inactivating knock-in mutation into the NPxY2 motif of LRP1 might influence the cell surface expression of LRP1 and NMDA receptors in primary cortical neurons. Here we demonstrate that a knock-in into the NPxY2 motif of LRP1 results in an increased surface expression of LRP1 and NR2B NMDA receptor subunit due to reduced endocytosis rates of LRP1 and the NR2B subunit in primary neurons derived from LRP1ΔNPxY2 animals. Furthermore, we demonstrate an altered phosphorylation pattern of S1480 and Y1472 in the NR2B subunit at the surface of LRP1ΔNPxY2 neurons, while the respective kinases Fyn and casein kinase II are not differently regulated compared with wild type controls. Performing co-immunoprecipitation experiments we demonstrate that binding of LRP1 to NR2B might be linked by PSD95, is phosphorylation dependent and this regulation mechanism is impaired in LRP1ΔNPxY2 neurons. Finally, we demonstrate hyperactivity and changes in spatial and reversal learning in LRP1ΔNPxY2 mice, confirming the mechanistic interaction in a physiological readout.

Conclusions

In summary, our data demonstrate that LRP1 plays a critical role in the regulation of NR2B expression at the cell surface and may provide a mechanistic explanation for the behavioral abnormalities detected in neuronal LRP1 knock-out animals reported earlier.

【 授权许可】

   
2013 Maier et al.; licensee BioMed Central Ltd.

【 图 表 】

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