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Electrophysiological measurements of synaptic connectivity and plasticity in the longitudinal dentate gyrus network from mouse hippocampal slices.

ABSTRACT: Longitudinal synaptic connections between dentate gyrus (DG) granule neurons in the hippocampus have been found to be correlated with increased anxiety. Here, we present a protocol to assess synaptic connectivity and plasticity in the longitudinal DG network. We detail the steps for (1) obtaining acute mouse hippocampal slices that contain longitudinal DG-DG connections, (2) measuring excitatory postsynaptic potentials using whole-cell patch clamp recording combined with two-photon microscopy and glutamate uncaging, and (3) assessing synaptic plasticity using extracellular field recording. For complete details on the use and execution of this protocol, please refer to Pak et al. (2022).¹.

SUBMITTER: Kwon YJ

PROVIDER: S-EPMC9852689 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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Electrophysiological measurements of synaptic connectivity and plasticity in the longitudinal dentate gyrus network from mouse hippocampal slices.

Kwon Yoon Ji YJ Pak Sojeong S Yang Sunggu S Yang Sungchil S

STAR protocols 20230111 1

Longitudinal synaptic connections between dentate gyrus (DG) granule neurons in the hippocampus have been found to be correlated with increased anxiety. Here, we present a protocol to assess synaptic connectivity and plasticity in the longitudinal DG network. We detail the steps for (1) obtaining acute mouse hippocampal slices that contain longitudinal DG-DG connections, (2) measuring excitatory postsynaptic potentials using whole-cell patch clamp recording combined with two-photon microscopy an ...[more]

PMID: 36633949

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Project description:The hippocampus is one of the most widely studied areas in the brain because of its important functional role in memory processing and learning, its remarkable neuronal cell plasticity, and its involvement in epilepsy, neurodegenerative diseases, and psychiatric disorders. The hippocampus is composed of distinct regions; the dentate gyrus, which comprises mainly granule neurons, and Ammon's horn, which comprises mainly pyramidal neurons, and the two regions are connected by both anatomic and functional circuits. Many different mRNAs and proteins are selectively expressed in the dentate gyrus, and the dentate gyrus is a site of adult neurogenesis; that is, new neurons are continually generated in the adult dentate gyrus. To investigate mRNA and protein expression specific to the dentate gyrus, laser capture microdissection is often used. This method has some limitations, however, such as the need for special apparatuses and complicated handling procedures. In this video-recorded protocol, we demonstrate a dissection technique for removing the dentate gyrus from adult mouse under a stereomicroscope. Dentate gyrus samples prepared using this technique are suitable for any assay, including transcriptomic, proteomic, and cell biology analyses. We confirmed that the dissected tissue is dentate gyrus by conducting real-time PCR of dentate gyrus-specific genes, tryptophan 2,3-dioxygenase (TDO2) and desmoplakin (Dsp), and Ammon's horn enriched genes, Meis-related gene 1b (Mrg1b) and TYRO3 protein tyrosine kinase 3 (Tyro3). The mRNA expressions of TDO2 and Dsp in the dentate gyrus samples were detected at obviously higher levels, whereas Mrg1b and Tyro3 were lower levels, than those in the Ammon's horn samples. To demonstrate the advantage of this method, we performed DNA microarray analysis using samples of whole hippocampus and dentate gyrus. The mRNA expression of TDO2 and Dsp, which are expressed selectively in the dentate gyrus, in the whole hippocampus of alpha-CaMKII+/- mice, exhibited 0.037 and 0.10-fold changes compared to that of wild-type mice, respectively. In the isolated dentate gyrus, however, these expressions exhibited 0.011 and 0.021-fold changes compared to that of wild-type mice, demonstrating that gene expression changes in dentate gyrus can be detected with greater sensitivity. Taken together, this convenient and accurate dissection technique can be reliably used for studies focused on the dentate gyrus.

Dataset Information

Electrophysiological measurements of synaptic connectivity and plasticity in the longitudinal dentate gyrus network from mouse hippocampal slices.

Publications

Electrophysiological measurements of synaptic connectivity and plasticity in the longitudinal dentate gyrus network from mouse hippocampal slices.

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