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Mechanical injury to neuronal/glial cultures in microplates: Role of NMDA receptors and pH in secondary neuronal cell death
Author(s) -
Mukhin A.G.,
Ivanova S.A.,
Allen J.W.,
Faden A.I.
Publication year - 1998
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/(sici)1097-4547(19980315)51:6<748::aid-jnr8>3.0.co;2-b
Subject(s) - nmda receptor , receptor , neuroscience , programmed cell death , chemistry , biology , microbiology and biotechnology , apoptosis , biochemistry
In vitro models of traumatic injury are useful adjuncts to animal models for studying mechanisms of post‐traumatic cell death. Here we describe a new in vitro model in which reproducible levels of injury are delivered by a punch device that produces 28 parallel cuts in individual wells of 96‐well microplates. Cell loss is measured by LDH assay or quantitative fluorometric assay for ethidium homodimer staining. Glial cultures show cell death restricted to the initial injury site, whereas neuronal/glial cultures demonstrate substantial spread of cell loss over time. We used this model to examine the role of pH and NMDA receptors in delayed post‐traumatic injury. NMDA receptor blockade by dizocilpine (MK‐801) or treatment with antisense oligodeoxynucleotides directed against NMDAR1 was neuroprotective. Decreased cell death was observed under acidic conditions whereas increased extracellular pH was associated with increased, MK‐801 sensitive cell loss. Advantages of our model include: reproducible trauma induction; rapid measurements of cell injury; and use of 96‐well microplates which reduce time and cost. This model appears to be well‐suited for the study of selected mechanisms of post‐traumatic neuronal injury as well as for screening potential neuroprotective agents. J. Neurosci. Res. 51:748–758, 1998. © 1998 Wiley‐Liss, Inc.