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Published on: February 27, 2018
Matrix metalloproteinase 9 regulates cell death following pilocarpine-induced seizures in the developing brain
Yvonne Hoehna1, Ortrud Uckermann, Hella Luksch
1Department of Pediatric Neurology, Children's Hospital, Medical Faculty Carl Gustav Carus, Technical University Dresden, Fetscherstrasse 74, 01307 Dresden, Germany.
Abstract:
Matrix metalloproteinases (MMPs) are involved in tissue repair, cell death and morphogenesis. We investigated the role of the gelatinases MMP-2 and MMP-9 in the pathogenesis of neuronal death induced by prolonged seizures in the developing brain. Seven-day-old rats, MMP-9 knockout mice and transgenic rats overexpressing MMP-9 received intraperitoneal injections of pilocarpine, 250 mg/kg, to induce seizures. After 6-72 h pups were sacrificed, tissue from different brain regions was isolated and expression of MMP-9 mRNA and protein was analyzed by real-time PCR or Western blot. Additionally, brains were fixed and processed for TUNEL-staining, immunohistochemistry and in situ zymography. We found increased numbers of TUNEL-positive cells 24 h after pilocarpine-induced seizures, most pronounced in cortical areas and the dentate gyrus, and less pronounced in thalamus. At 6-24 h, MMP-9 mRNA levels showed significant elevation compared to sham-treated controls; this effect resolved by 48 h, whereas MMP-2 mRNA levels remained stable. Cortical gelatinolytic activity, monitored by in situ zymography, was enhanced following pilocarpine-induced seizures. The MMP inhibitor GM 6001 ameliorated cell death following pilocarpine-induced seizures in infant rats. MMP-9 knockout mice were less susceptible to seizure-induced brain injury. Transgenic rats overexpressing MMP-9 were equally susceptible to seizure-induced brain injury as wild type rats. Our results suggest a significant contribution of MMP-9 to cell death after pilocarpine-induced seizures in the developing brain. As indicated by Western blot analysis, MMP-9 activation may be linked to activation of the Erk/CREB-pathway. The findings implicate involvement of MMP-9 in the pathophysiology of brain injury following seizures in the developing brain.
Insights
Matrix metalloproteinases-9 (MMP-9) contribute to neuronal death following prolonged seizures in developing brains. Inhibiting MMP-9 reduced injury, while MMP-9 knockout mice showed less susceptibility to seizure-induced brain damage.
Area of Science:
- Neuroscience
- Biochemistry
- Developmental Biology
Background:
- Matrix metalloproteinases (MMPs) play roles in tissue repair and cell death.
- The specific involvement of gelatinases MMP-2 and MMP-9 in seizure-induced neuronal death in developing brains remains unclear.
Purpose of the Study:
- To investigate the role of MMP-2 and MMP-9 in neuronal death following prolonged seizures in the developing brain.
Main Methods:
- Pilocarpine was used to induce seizures in 7-day-old rats, MMP-9 knockout mice, and transgenic rats overexpressing MMP-9.
- Analysis included real-time PCR, Western blot, TUNEL staining, immunohistochemistry, and in situ zymography.
- MMP inhibitor GM 6001 was administered to assess its effect on cell death.
Main Results:
- Pilocarpine-induced seizures increased TUNEL-positive cells, particularly in cortical areas and the dentate gyrus.
- MMP-9 mRNA levels significantly elevated post-seizure, while MMP-2 mRNA remained stable.
- Enhanced gelatinolytic activity was observed in the cortex; MMP-9 knockout mice exhibited reduced susceptibility to injury.
Conclusions:
- MMP-9 significantly contributes to cell death after pilocarpine-induced seizures in the developing brain.
- MMP-9 activation may be linked to the Erk/CREB pathway.
- MMP-9 is implicated in the pathophysiology of brain injury following seizures in developing brains.
