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Regulation of Dendrite Arborization by Substrate Stiffness is Mediated by Glutamate Receptors

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Abstract

Brain injury or disease can initiate changes in local or global stiffness of brain tissue. While stiffness of the extracellular environment is known to affect the morphology and function of many cell types, little is known about how the dendrites of neurons respond to changes in brain stiffness. To assess how extracellular stiffness affects dendrite morphology, we took biomaterial and biomedical engineering approaches. We cultured mixed and pure hippocampal neurons on hydrogels composed of polyacrylamide (PA) of varying stiffnesses to mimic the effects of extracellular matrix stiffness on dendrite morphology. The majority of investigations of cortical and spinal cord neurons on soft hydrogels examined branching at early time points (days in vitro (DIV) 2–7), an important distinction from our study, where we include later time points that encompass the peak of branching (DIV 10–12). At DIV 12, dendrite branching was altered by stiffness for both pure and mixed neuronal cultures. Furthermore, we treated hippocampal cultures with glutamate receptor antagonists and with astrocyte-conditioned media. Blocking AMPA and NMDA receptors affected the changes in dendrite branching seen at varying rigidities. Moreover, extracellular factors secreted by astrocytes also change dendrite branching seen at varying rigidities. Thus, astrocytes and ionotropic glutamate receptors contribute to mechanosensing.

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Abbreviations

PA:

Polyacrylamide

AMPA:

α-Amino-3-hydroxyl-5-methyl-4-isoxazole-propionate

NMDA:

N-Methyl-d-aspartic acid

DIV:

Days in vitro

APV:

dl-2-Amino-5-phosphonovalerate acid

CNQX:

6-Cyano-7-nitroquinoxaline-2,3-dione

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Acknowledgments

This work was funded by grants from National Science Foundation IBN-0548543 and IBN-0919747; March of Dimes 1-FY08-464; New Jersey Commission on Brain Injury Research, #09-3209-BIR-E-2; American Heart Association 0555801T. MLP was supported by the IGERT Program on Biointerfaces, NSF Grant DGE-0333196 and Louis Bevier Dissertation Fellowship. CGL was supported by Biotechnology Fellowship, Grant 5T32GM008339 from National Institute of General Medical Sciences. We would like to thank Dr. Penelope Georges for her scientific guidance and support.

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Authors and Affiliations

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

    Michelle L. Previtera, Christopher G. Langhammer & Bonnie L. Firestein

  2. Graduate Program in Molecular Biosciences, Rutgers University, 604 Allison Rd, Piscataway, NJ, 08854, USA

    Michelle L. Previtera

  3. Graduate Program in Biomedical Engineering, Rutgers University, 599 Taylor Rd, Piscataway, NJ, 08854, USA

    Christopher G. Langhammer

  4. Department of Biomedical Engineering, Rutgers University, 604 Allison Rd, Piscataway, NJ, 08854, USA

    Noshir A. Langrana & Bonnie L. Firestein

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  1. Michelle L. Previtera
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  4. Bonnie L. Firestein
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Correspondence to Bonnie L. Firestein.

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Associate Editor Jennifer West oversaw the review of this article.

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Previtera, M.L., Langhammer, C.G., Langrana, N.A. et al. Regulation of Dendrite Arborization by Substrate Stiffness is Mediated by Glutamate Receptors. Ann Biomed Eng 38, 3733–3743 (2010). https://doi.org/10.1007/s10439-010-0112-5

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