Elsevier

Brain Research

Volume 490, Issue 1, 19 June 1989, Pages 110-125
Brain Research

Development and selective neurodegeneration in cell cultures from different hippocampal regions

https://doi.org/10.1016/0006-8993(89)90436-8Get rights and content

Abstract

Previous studies have shown that pyramidal neurons in hippocampal regions CA1 and CA3 are selectively vulnerable in several neurodegenerative disorders and that a subpopulation of pyramidal neurons in cell cultures of embryonic hippocampus are sensitive to glutamate neurotoxicity. In order to determine whether the patterns of cell loss seen in situ correlate with intrinsic differences in neuronal sensitivities to glutamate-induced denegeration acquired during development, we characterized cultures established from different regions of postnatal rat hippocampus and then examined neuronal sensitivity to glutamate. Tissue corresponding to the dentate gyrus (DG) and regions CA1, CA2, and CA3 of Ammon's horn was removed by microdissection from transverse hippocampal slices and was used to establish cultures of dissociated cells. Cultures from all 4 regions contained 3 major morphological classes of neurons; pyramidal-like, bipolar and stellate. Pyramidal-like neurons comprised the majority of neurons in all cultures; these neurons extended one long and branching axon, and one or more short dendrites. Immunocytochemistry showed that all neurons possessed high levels of glutamate-like and γ-aminobutyric acid (GABA)-like immunoreactivity when grown in isolation. In contrast, when bipolar and pyramidal neurons were cultured in contact with glial cells, glutamate and GABA immunoreactivity were selectively reduced in the bipolar and pyramidal cells, respectively, suggesting that cell interactions influence neurotransmitter phenotype. Subpopulations of hippocampal neurons from each hippocampal region were vulnerable to glutamate-induced neurotoxicity. Bipolar and stellate cells were resistant to glutamate, while pyramidal-like neurons showed varying degrees of sensitivity to glutamate depending upon which region they were taken from. Experiments with specific glutamate receptor agonists and antagonists demonstrated that both non N-methyl-d-spartic acid (NMDA) receptors and NMDA receptors mediated glutamate-induced degeneration. There were clear differences in the vulnerability of the pyramidal-like neuron populations in cultures from the different hippocampal regions. The rank order of the vulnerability of pyramidal-like neurons to glutamate-induced neurodegeneration between regions in culture was: <CA2<CA3<CA1. This pattern of selective vulnerability in cell culture corresponds directly to the pattern of selective cell loss seen in situ in Alzheimer's disease, epilepsy, and stroke suggesting that intrinsic neuronal differences in glutamate sensitivity may be involved in these disorders.

References (68)

  • MattsonM.P.

    Neurotransmitters in the regulation of neuronal cytoarchitecture

    Brain Res. Rev.

    (1988)
  • MattsonM.P. et al.

    Isolated hippocampal neurons in cryopreserved long-term cultures: development of neuroarchitecture and sensitivity to NMDA

    Int. J. Dev. Neurosci.

    (1988)
  • MattsonM.P. et al.

    Interactions of entorhinal axons and target hippocampal neurons: a role of glutamate in the development of hippocampal circuitry

    Neuron

    (1988)
  • McCobbD.P. et al.

    Interactive effects of serotonin and acetylcholine on neurite elongation

    Neuron

    (1988)
  • ProbstA. et al.

    Neuritic plaques in senile dementia of the Alzheimer type: a golgi analysis in the hippocampal region

    Brain Research

    (1983)
  • RobainO. et al.

    GABAergic neurons of the hippocampus: development in homotopic graft and in dissociated cell cultures

    Neuroscience

    (1987)
  • SawadaS. et al.

    Selective activation of synapses near the tip of drug-ejecting microelectrode, and effects of antagonists of excitatory amino acids in the hippocampus

    Brain Research

    (1983)
  • SchwarczR. et al.

    Excitotoxic models for neurodegenerative disorders

    Life Sci.

    (1984)
  • SloviterR.S.

    ‘Epileptic’ brain damage in rats induced by sustained electrical stimulation of the perforant path. I. Acute electrophysiological and light microscopic studies

    Brain Res. Bull.

    (1983)
  • Van HoesenG.W. et al.

    Cell-specific pathology in neural systems of the temporal lobe in Alzheimer's disease

    Prog. Brain Res.

    (1986)
  • WatkinsJ.C. et al.

    Agonists and antagonists for excitatory amino acid receptors

    Trends Neurosci.

    (1987)
  • BankerG.A. et al.

    Further observations on hippocampal neurons in dispersed cell culture

    J. Comp. Neurol.

    (1979)
  • BlackI.B. et al.

    Neurotransmitter plasticity at the molecular level

    Science

    (1984)
  • CaceresA. et al.

    Differential subcellular localization of tubulin and the microtubule-associated protein MAP2 in brain tissue as revealed by immunocytochemistry with monoclonal hybridoma antibodies

    J. Neurosci.

    (1984)
  • CaceresA. et al.

    Immunocytochemical localization of microtubule-associated protein 2 during the development of hippocampal neurons in culture

    J. Neurosci.

    (1986)
  • ChouS.M.

    Pathognomy of intraneuronal inclusion in ALS

  • CollinridgeG.I. et al.

    The antagonism of amino acid-induced excitations of rat hippocampal CA1 neurons in vitro

    J. Physiol. (Lond.)

    (1983)
  • CollinridgeG.I. et al.

    Excitatory amino acids in synaptic transmission in the Schaffer collateral-commissural pathway of the rat hippocampus

    J. Physiol. (Lond.)

    (1983)
  • CotmanC.W.

    Synaptic Plasticity

  • CotmanC.W. et al.

    Anatomical organization of excitatory amino acid receptors and their pathways

    Trends Neurosci.

    (1987)
  • CowanW.M. et al.

    Further observations on the development of the dentate gyrus

  • CoyleJ.T. et al.

    Lesion of striatal neurons with kainic acid provides a model for Huntington's chorea

    Nature (Lond.)

    (1976)
  • CoyleJ.T. et al.

    Excitatory amino acid neurotoxins: selectivity, specificity, and mechanisms of action

    Neurosci. Res. Prog. Bull.

    (1981)
  • DamA.M.

    Epilepsy and neuron loss in the hippocampus

    Epilepsia

    (1980)
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