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Interaction Between CRIPT and PSD-95 Is Required for Proper Dendritic Arborization in Hippocampal Neurons

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Abstract

CRIPT, the cysteine-rich PDZ-binding protein, binds to the third PDZ domain of PSD-95 (postsynaptic density protein 95) family proteins and directly binds microtubules, linking PSD-95 family proteins to the neuronal cytoskeleton. Here, we show that overexpression of a full-length CRIPT leads to a modest decrease, and knockdown of CRIPT leads to an increase in dendritic branching in cultured rat hippocampal neurons. Overexpression of truncated CRIPT lacking the PDZ domain-binding motif, which does not bind to PSD-95, significantly decreases dendritic arborization. Conversely, overexpression of a full-length CRIPT significantly increases the number of immature and mature dendritic spines, and this effect is not observed when CRIPT∆PDZ is overexpressed. Competitive inhibition of CRIPT binding to the third PDZ domain of PSD-95 with PDZ3-binding peptides resulted in differential effects on dendritic arborization based on the origin of respective peptide sequence. These results highlight multifunctional roles of CRIPT during development and underscore the significance of the interaction between CRIPT and the third PDZ domain of PSD-95.

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Funding

This work was funded by National Science Foundation grant IOS-1353724 and New Jersey Commission on Brain Injury Research grant #CBIR14IRG019 to B.L.F. A.O. was supported by National Institutes of Health Biotechnology Training Grant (T32 GM008339-20) and a predoctoral fellowship from the New Jersey Commission on Brain Injury Research #CBIR19FEL018. HM was supported by National Institutes of Health IRACDA (Institutional Research and Career Development Award) INSPIRE (IRACDA2K12GM093854-07A1). MS received funding from Norris Cotton Cancer Center, Geisel School of Medicine at Dartmouth.

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  1. Anton Omelchenko and Harita Menon contributed equally to this work.

Authors and Affiliations

  1. Department of Cell Biology and Neuroscience, Rutgers University, 604 Allison Road, Piscataway, NJ, 08854-8082, USA

    Anton Omelchenko, Harita Menon, Sarah G. Donofrio, Gaurav Kumar, Joshua Roshal, Eduardo R. Martinez-Montes & Bonnie L. Firestein

  2. Neuroscience Graduate Program, Rutgers University, 604 Allison Road, Piscataway, NJ, 08854-8082, USA

    Anton Omelchenko

  3. Geisel School of Medicine, Department of Medical Education and Norris Cotton Cancer Center, Dartmouth College, Lebanon, NH, 03756, USA

    Heidi M. Chapman, Tiffany L. Wang & Mark R. Spaller

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  1. Anton Omelchenko
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Contributions

HM, MS, and BLF designed the experiments. HM, SD, GK, JR, HMC, ERM-M, and TW executed the experiments. HM, SD, GK, JR, and ERM-M performed the biochemical experiments and analysis of CRIPT overexpression and treatment of peptides on cultured neurons. HMC and TW synthesized the peptides. AO and BLF wrote the manuscript with input from HM and MS. AO analyzed all data. MS supervised peptide production and optimization, and BLF supervised the project.

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Correspondence to Bonnie L. Firestein.

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Omelchenko, A., Menon, H., Donofrio, S.G. et al. Interaction Between CRIPT and PSD-95 Is Required for Proper Dendritic Arborization in Hippocampal Neurons. Mol Neurobiol 57, 2479–2493 (2020). https://doi.org/10.1007/s12035-020-01895-5

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