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Diltiazem-induced Neuroprotection in Glutamate Excitotoxicity and Ischemic Insult of Retinal Neurons

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Abstract

Purpose: Cell death is often related to an abnormal increase in Ca2+ flux. In the retina, Ca2+ channels are mainly from the L-type that do not inactivate with time. Under excitotoxic and ischemic conditions, their continuous activation may therefore contribute significantly to the lethal Ca2+ influx. To assess this hypothesis, the Ca2+ channel blocker, diltiazem, was applied in excitotoxic and ischemic conditions. Methods: To induce excitotoxicity, retinal cell cultures from newborn rats were incubated with glutamate. The toxicity of glutamate was quantified by neuronal immunostaining with an antibody directed against the neuron specific enolase. Glutamate receptor function in vitro was assessed in pig retinal cell cultures by patch clamp recording. Retinal ischemia was induced by raising the intraocular pressure in adult rats. Retinal cell loss was quantified on retinal sections by measuring nuclear cell densities. Results: In retinal cell culture, glutamate application induced a major cell loss. This cell loss was attributed to glutamate excitotoxicity because glutamate receptor blockers like MK-801 and CNQX increased significantly neuronal survival. MK-801 and CNQX, which block NMDA and AMPA/Kainate receptors, respectively, had additive effects. Expression of AMPA/Kainate glutamate receptors in mixed adult retinal cell cultures was attested by patch clamp recording. In newborn rat retinal culture, glutamate excitotoxicity was significantly reduced by addition of the L-type Ca2+ channel blocker, diltiazem. In in vivo experiments, the increase in ocular pressure induced a decrease in cell number in the inner nuclear and ganglion cell layers. When animals received diltiazem injections, the ischemic treatment induced a less severe reduction in retinal cells; this neuroprotection was statistically significant in the ganglion cell layer. Conclusion: These results are consistent with previous studies suggesting that Ca2+ channel activation contributes to retinal cell death following either glutamate excitotoxicity or retinal ischemia. Under both conditions, the L-type Ca2+ channel blocker, diltiazem, can limit cell death. These results extend the potential application of diltiazem in retinal neuroprotection to retinal pathologies involving glutamate excitotoxicity and ischemia.

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Authors and Affiliations

  1. Laboratoire de Physiopathologie Cellulaire et Moléculaire de la Rétine, UPMC, INSERM U-592, Hôpital Saint-Antoine, Bâtiment Kourilsky, Paris, France

    Géraldine Vallazza-Deschamps, Céline Fuchs, David Cia, Luc-Henri Tessier, José A. A. Sahel, Henri Dreyfus & Serge Picaud

  2. Centre Hospitalier National d’Ophtalmologie des Quinze-Vingts, Paris, France

    José A. A. Sahel

  3. Fondation Ophtalmologique Adolphe de Rothschild, Paris, France

    José A. A. Sahel & Serge Picaud

  4. AP-HP, Paris, France

    Serge Picaud

Authors
  1. Géraldine Vallazza-Deschamps
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  2. Céline Fuchs
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  3. David Cia
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  4. Luc-Henri Tessier
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  5. José A. A. Sahel
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  6. Henri Dreyfus
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  7. Serge Picaud
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Correspondence to Serge Picaud.

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Vallazza-Deschamps, G., Fuchs, C., Cia, D. et al. Diltiazem-induced Neuroprotection in Glutamate Excitotoxicity and Ischemic Insult of Retinal Neurons. Doc Ophthalmol 110, 25–35 (2005). https://doi.org/10.1007/s10633-005-7341-1

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