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Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome
Published on: June 10, 2013
MicroRNA-34a negatively regulates anesthesia-induced hippocampal apoptosis and memory impairment through FGFR1
Xiu-Li Jiang1, Bo-Xiang Du1, Jie Chen2
1Department of Anesthesiology, The Second Affiliated Hospital of Nantong University Nantong 226001, Jiangsu Province, China.
Background:
Mounting evidence has shown the toxic effects of anesthesia to neonatal hippocampus. We used an in vivo mouse model to explore the role of microRNA 34a (miR-34a) in regulating anesthesia-induced hippocampal neurotoxicity.
Methods:
One-month old C57/BL6 mice received daily intraperitoneal injection of anesthesia (ketamine, 50 mg/kg) for 7 days. One day after, apoptosis was evaluated by TUNEL staining in hippocampal CA1 region, and expression level of miR-34a assessed by real-time quantitative PCR (qPCR). Hippocampal miR-34a was then down-regulated through lentivirus mediated cortical injection prior to anesthesia. The effects of inhibiting hippocampal miR-34a on anesthesia-induced hippocampal apoptosis and memory impairment were further investigated by TUNEL staining and Morris water maze (MWM) test. The predicted molecular target of miR-34a, fibroblast growth factor receptor 1 (FGFR1) was down-regulated in hippocampus through siRNA-mediated cortical injection and its effect on hippocampal apoptosis was also examined.
Results:
Anesthesia caused severe apoptosis among hippocampal CA1 neurons and upregulated hippocampal miR-34a. On the other hand, lentivirual inhibition of miR-34a protected anesthesia-induced hippocampal apoptosis and memory impairment. Luciferase essay demonstrated FGFR1 was directly regulated by miR-34a in hippocampus. siRNA-induced FGFR1 downregulation further exaggerated anesthesia-induced apoptosis in hippocampus.
Conclusions:
Overall, we showed that miR-34a negatively modulated anesthesia-induced hippocampal neurotoxicity.
Insights
Anesthesia harms the neonatal hippocampus. MicroRNA 34a (miR-34a) plays a key role in this neurotoxicity, with its inhibition protecting against damage and memory loss.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Anesthesia is known to cause toxic effects on the neonatal hippocampus.
- The precise molecular mechanisms underlying anesthesia-induced hippocampal neurotoxicity require further elucidation.
Purpose of the Study:
- To investigate the role of microRNA 34a (miR-34a) in anesthesia-induced hippocampal neurotoxicity.
- To explore the therapeutic potential of modulating miR-34a in preventing anesthesia-related brain injury.
Main Methods:
- An in vivo mouse model was utilized, administering anesthesia (ketamine) daily for seven days.
- Apoptosis was assessed using TUNEL staining, and miR-34a levels were quantified via qPCR.
- miR-34a was inhibited using lentivirus, and fibroblast growth factor receptor 1 (FGFR1) was targeted with siRNA to evaluate their effects on neurotoxicity and memory.
Main Results:
- Anesthesia exposure led to significant hippocampal CA1 neuron apoptosis and increased miR-34a expression.
- Inhibition of miR-34a via lentivirus administration attenuated anesthesia-induced apoptosis and improved memory function.
- FGFR1 was identified as a direct target of miR-34a, and its downregulation exacerbated anesthesia-induced apoptosis.
Conclusions:
- MicroRNA 34a (miR-34a) acts as a negative regulator of anesthesia-induced hippocampal neurotoxicity.
- Targeting miR-34a presents a potential therapeutic strategy to mitigate anesthesia-related neurological damage in neonates.
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