Association of c‐ fos mRNA expression and excitotoxicity in primary cultures of mouse neocortical and cerebellar neurons

Association of c‐ fos mRNA expression and excitotoxicity in primary cultures of mouse... The effect of excitatory amino acids (EAAs) on c‐fos mRNA expression was studied in primary cultures of mouse cerebellar granule cells and in neocortical neurons after 2 and 7 days in vitro (div). In cultured granule cells at 2 and 7 div, and in cortical neurons at 2 div, exposure to low levels (≤10 μM) of a variety of EAAs (viz. glutamate (Glu), S‐sulpho‐L‐cysteine (SC), N‐methyl‐D‐aspartate (NMDA), α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazole (AMPA), and kainate (KA)) resulted in a transient increase in the level of c‐fos mRNA which peaked at 30 min but returned to a basal level by 120 min. However, exposure of granule cells (7 div) to high levels (250 μM) of Glu, NMDA, KA, SC and of cortical neurons (7 div) to high levels (250 μM) of Glu, NMDA, KA, SC, or AMPA and to low levels (≤10 μM) of Glu and AMPA resulted in a delay in c‐fos mRNA induction but a subsequent, progressive increase that was sustained for at least 240 min. Furthermore, this effect was accompanied by a dose‐related increase in the release of the cytosolic enzyme, lactate dehydrogenase, used as an indicator of excitotoxicity. A ratio (Q240/30) for the steady‐state levels of c‐fos mRNA after 30 min and 240 min of exposure to EAAs was determined which showed that Q240/30 >2 correlated reproducibly with excitotoxic cell death, whereas a ratio of ≤1 correlated with a nonexcitotoxic event. In both cell types at 7 div, coadministration of the selective NMDA receptor antagonist, DL(±)‐2‐amino‐5‐phosphonopentanoic acid (APV) with cytotoxic levels of Glu 1) protected against EAA‐induced neurotoxicity and 2) exhibited a transient c‐fos mRNA expression (Q240/30 values ≈1). In contrast, the AMPA/KA receptor antagonist, 6‐cyano‐7‐nitroquinoxaline‐2,3‐dione (CNQX), provided no protection against excitotoxicity and had no significant effect on the Glu‐induced delay in c‐fos mRNA expression. These results suggest that the Q240/30 c‐fos mRNA ratio may 1) be used as a predictive index for excitotoxic neuronal death, 2) provide information on the identity of the receptor subtype mediating excitotoxicity in different brain cell types, and 3) aid in establishing the role of excitotoxicity during the development of neurons in vitro. J. Neurosci. Res. 48:533–542, 1997. © 1997 Wiley‐Liss Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neuroscience Research Wiley

Association of c‐ fos mRNA expression and excitotoxicity in primary cultures of mouse neocortical and cerebellar neurons

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Publisher
Wiley
Copyright
Copyright © 1997 Wiley‐Liss, Inc.
ISSN
0360-4012
eISSN
1097-4547
D.O.I.
10.1002/(SICI)1097-4547(19970615)48:6<533::AID-JNR6>3.0.CO;2-B
Publisher site
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Abstract

The effect of excitatory amino acids (EAAs) on c‐fos mRNA expression was studied in primary cultures of mouse cerebellar granule cells and in neocortical neurons after 2 and 7 days in vitro (div). In cultured granule cells at 2 and 7 div, and in cortical neurons at 2 div, exposure to low levels (≤10 μM) of a variety of EAAs (viz. glutamate (Glu), S‐sulpho‐L‐cysteine (SC), N‐methyl‐D‐aspartate (NMDA), α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazole (AMPA), and kainate (KA)) resulted in a transient increase in the level of c‐fos mRNA which peaked at 30 min but returned to a basal level by 120 min. However, exposure of granule cells (7 div) to high levels (250 μM) of Glu, NMDA, KA, SC and of cortical neurons (7 div) to high levels (250 μM) of Glu, NMDA, KA, SC, or AMPA and to low levels (≤10 μM) of Glu and AMPA resulted in a delay in c‐fos mRNA induction but a subsequent, progressive increase that was sustained for at least 240 min. Furthermore, this effect was accompanied by a dose‐related increase in the release of the cytosolic enzyme, lactate dehydrogenase, used as an indicator of excitotoxicity. A ratio (Q240/30) for the steady‐state levels of c‐fos mRNA after 30 min and 240 min of exposure to EAAs was determined which showed that Q240/30 >2 correlated reproducibly with excitotoxic cell death, whereas a ratio of ≤1 correlated with a nonexcitotoxic event. In both cell types at 7 div, coadministration of the selective NMDA receptor antagonist, DL(±)‐2‐amino‐5‐phosphonopentanoic acid (APV) with cytotoxic levels of Glu 1) protected against EAA‐induced neurotoxicity and 2) exhibited a transient c‐fos mRNA expression (Q240/30 values ≈1). In contrast, the AMPA/KA receptor antagonist, 6‐cyano‐7‐nitroquinoxaline‐2,3‐dione (CNQX), provided no protection against excitotoxicity and had no significant effect on the Glu‐induced delay in c‐fos mRNA expression. These results suggest that the Q240/30 c‐fos mRNA ratio may 1) be used as a predictive index for excitotoxic neuronal death, 2) provide information on the identity of the receptor subtype mediating excitotoxicity in different brain cell types, and 3) aid in establishing the role of excitotoxicity during the development of neurons in vitro. J. Neurosci. Res. 48:533–542, 1997. © 1997 Wiley‐Liss Inc.

Journal

Journal of Neuroscience ResearchWiley

Published: Jun 15, 1997

References

  • Translational suppression of calpain I reduces NMDA‐induced spectrin proteolysis and pathophysiology in cultured hippocampal slices
    Bednarski, Bednarski; Vanerklish, Vanerklish; Gall, Gall; Saido, Saido; Bahr, Bahr; Lynch, Lynch
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  • Cytotoxic actions and effects on intracellular Ca 2+ and cGMP concentrations of sulphur‐containing excitatory amino acids in cultured cerebral cortical neurons
    Frandsen, Frandsen; Schousboe, Schousboe; Griffiths, Griffiths
  • Calcium regulation of gene expression: the mode of entry matters
    Gallin, Gallin; Greenberg, Greenberg
  • Neuronal expression of AP‐1 proteins in excitotoxic‐neurodegenerative disorders and following nerve fiber lesions
    Gass, Gass; Herdegen, Herdegen
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    Karin, Karin; Hunter, Hunter
  • Calcium‐mediated mechanisms in chemically induced cell death
    Nicotera, Nicotera; Bellomo, Bellomo; Orrenius, Orrenius

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