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A Standardized Protocol for Stereotaxic Intrahippocampal Administration of Kainic Acid Combined With Electroencephalographic Seizure Monitoring in Mice

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A Standardized Protocol for Stereotaxic Intrahippocampal Administration of Kainic Acid Combined With Electroencephalographic Seizure Monitoring in Mice

Pascal Bielefeld et al. Front Neurosci.

Abstract

Lack of scientific reproducibility is a growing concern and weak experimental practices may contribute to irreproducibility. Here, we describe an optimized and versatile protocol for stereotaxic intrahippocampal administration of Kainic Acid (KA) in mice with a C57Bl6 background. In this protocol, KA administration is combined with in vivo recording of neuronal activity with wired and wireless setups. Following our protocol, KA administration results in a robust dose-dependent induction of low-level epileptiform activity or Status Epilepticus (SE) and induces previously characterized hallmarks of seizure-associated pathology. The procedure consists of three main steps: Craniotomy, stereotaxic administration of KA, and placement of recording electrodes in intrahippocampal, and subdural locations. This protocol offers extended possibilities compared to the systemic administration of KA, as it allows the researcher to accurately regulate the local dose of KA and resulting seizure activity, and permits the use and study of convulsive and non-convulsive KA doses, resulting in higher reproducibility and lower inter-individual variability and mortality rates. Caution should be taken when translating this procedure to different strains of mice as inter-strain sensitivity to KA has been described before. The procedure can be performed in ~1 h by a trained researcher, while intrahippocampal administration of KA without placing recording electrodes can be done in 25 min, and can be easily adapted to the titrated intrahippocampal administration of other drugs.

Keywords: EEG; adult hippocampal neurogenesis; epilepsy; intrahippocampal administration; kainic acid; temporal lobe.

Figures

Figure 1
Figure 1
Kainic Acid dose-dependent effects on gliosis (A) and granule cell dispersion (B), 28 days post KA administration. Immunohistochemistry against GFAP reveals clear dose-dependent gliosis effects, becoming apparent at 2.2 mM KA, which coincides with the presence of epileptic seizures. A Nissl staining to assess granule cell dispersion only shows granule cell dispersion when administering 20 mM KA, while the seizure-inducing 2.2 mM shows no sign of dispersion. Interestingly, for both gliosis and granule dispersion, the effects are limited to the ipsilateral hemisphere.
Figure 2
Figure 2
Hardware needed for intrahippocampal administration of KA and a guideline including crucial steps in the surgery procedure for electrode implantation and EEG recordings. (A) The stereotaxic setup in which the NanoJect is placed. (B) Close-up image of the injection capillary. (C) Example of a recording electrode as used for the subdural wireless EEG recordings. (D) Location of the burr holes in the skull as used for the wireless EEG electrode implantation. (E) Placement of the subdural recording electrodes in the burr holes. (F) Attaching the recording electrodes to the skull with dental cement. (G) Covering the recording electrodes in dental cement and attachment of the pin connector. (H) Example of a mouse carrying a wireless recording device. (I) Example of a mouse in the wired EEG recording setup.
Figure 3
Figure 3
Example traces of noise-free and noisy EEG recordings and Faraday cage design. (A) Example trace of a noise-free recording showing small deviations from baseline activity. (B) Example trace of a noisy recording showing significant deviations from baseline activity to an extent that may overlap with changes from baseline in the noise-free recording. (C) Example of a Faraday cage used to prevent noisy EEG recordings.

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